Body-consolidating and life-prolonging pill for improving frequent micturition at night and preparation method thereof

Abstract

The invention relates to the technical field of traditional Chinese medicines, in particular to a foundation-consolidating and life-prolonging pill for improving frequent urination at night and a preparation method of the foundation-consolidating and life-prolonging pill. The invention discloses a foundation strengthening and life prolonging pill for improving frequent micturition at night. The traditional Chinese medicine composition comprises the following components in parts by weight: 3-12 parts of ginseng, 5-15 parts of Chinese yam, 3-12 parts of dogwood, 2-14 parts of akebiaquinata, 3-10 parts of exocarpium benincasae, 2-10 parts of pericarpium arecae, 1-8 parts of semen boitae, 3-12 parts of radix achyranthis bidentatae, 1-7 parts of deer-horn glue, 1-7 parts of tortoise-plastron glue, 5-16 parts of pyrrosia lingua, 0.5-5 parts of liquorice, 3-10 parts of morinda officinalis, 4-15 parts of mantis egg-case, 3-10 parts of fructus alpiniae oxyphyllae, 6-20 parts of fructus rosae laevigatae, 6-20 parts of raspberry and 6-20 parts of The foundation-consolidating and life-prolonging pill does not cause pathological damage to drug metabolism visceral liver tissue, urine metabolism and regulation of visceral kidney, adrenal gland and bladder tissue. According to a high-dose group of the foundation strengthening and life prolonging pills, the urination latency period can be remarkably prolonged, the urine volume and weight ratio and the total excretion rate can be reduced, the urination latency period of the foundation strengthening and life prolonging pill is prolonged to 47.88 min from 34.38 min, the urine volume and weight ratio is reduced to 29.86 mL / kg from 44.81 mL / kg, and the total excretion rate is reduced to 62.86% from 81.37%.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicines, and in particular to a Guben Yanling pill for improving nocturnal urination and a preparation method thereof. Background Art

[0002] Frequent urination at night is a common symptom among middle-aged and elderly people, often associated with kidney qi deficiency, bladder qi dysfunction, or spleen and kidney yang deficiency, severely impacting sleep quality and well-being. Currently, Western medicine primarily treats this condition with alpha-blockers or antidiuretic hormones, but long-term use can lead to side effects such as electrolyte imbalances and headaches. Traditional Chinese medicine formulas, such as Suquan Pills (Alpinia oxyphylla, Linderae sibiricum, and Chinese yam) and Jinkui Shenqi Pills, offer unique advantages in improving nocturnal urination by targeting multiple pathways. However, these formulas offer limited efficacy in treating complex syndromes characterized by damp-heat or qi and blood deficiency. Modern medicine attributes its pathogenesis to factors such as reduced bladder capacity, weakened urinary regulation, and irregular adrenal hormone secretion. Traditional Chinese medicine, however, attributes this condition to "weak kidney qi," "weakened life gate fire," and "bladder dysfunction."

[0003] Traditional Guben Yanling Pills are a tonic Chinese patent medicine with benefits such as tonifying qi and blood, strengthening the spleen and kidneys, and calming the mind. However, their formula focuses on "prolonging life" and "strengthening the body" rather than "strengthening and stopping nocturnal urination." While existing Guben Yanling Pills have some effect on relieving frequent urination at night, their efficacy is less robust, with limitations such as a non-focused formula and insufficient astringency.

[0004] Therefore, there is an urgent need to develop a traditional Chinese medicine compound that can nourish the kidney and consolidate essence, eliminate dampness and clear turbidity, strengthen the spleen and replenish qi, and by optimizing the formula structure, enhance its targeting of reducing urine and stopping nocturnal emission, and warming and nourishing kidney yang, so as to more comprehensively improve the symptoms of frequent urination at night, thereby developing a compound traditional Chinese medicine preparation that is more suitable for people with frequent urination at night. Summary of the Invention

[0005] In response to the deficiencies in the above-mentioned prior art, the present invention provides a Guben Yanling Pill for improving nocturnal frequent urination and a preparation method thereof. By deleting medicinal materials unrelated to the main symptoms on the basis of the original formula and adding a variety of Chinese medicinal materials with the effect of reducing urine flow and stopping nocturnal emission, a composition with synergistic enhancement and strong targeting is formed. At the same time, a preparation method and animal experimental verification thereof are provided.

[0006] In order to solve the above technical problems, the technical solution adopted by the present invention is:

[0007] A Guben Yanling Pill for improving frequent nocturnal urination is characterized by being prepared from the following Chinese medicinal raw materials: ginseng, yam, cornus officinalis, akebia, wax gourd peel, dried tangerine peel, cypress seed, achyranthes bidentata, antler glue, tortoise shell glue, pyrrosia, licorice, Morinda officinalis, mulberry silkworm cocoon, Alpinia oxyphylla, Rosa laevigata, raspberry, and cat's whiskers.

[0008] Preferably, the Guben Yanling Pills for improving frequent urination at night are prepared from the following Chinese medicinal raw materials, calculated by weight: 3-12 parts of ginseng, 5-15 parts of yam, 3-12 parts of cornus officinalis, 2-14 parts of akebia, 3-10 parts of wax gourd peel, 2-10 parts of dried tangerine peel, 1-8 parts of cypress seed, 3-12 parts of Achyranthes bidentata, 1-7 parts of antler glue, 1-7 parts of tortoise shell glue, 5-16 parts of pyrrosia, 0.5-5 parts of licorice, 3-10 parts of Morinda officinalis, 4-15 parts of mulberry silkworm cocoon, 3-10 parts of Alpinia oxyphylla, 6-20 parts of Rosa laevigata, 6-20 parts of raspberry, and 6-20 parts of cat's whiskers.

[0009] Furthermore, the Guben Yanling Pills for improving frequent nocturnal urination are prepared from the following Chinese medicinal raw materials, calculated by weight: 4-8 parts of ginseng, 8-12 parts of yam, 4-10 parts of cornus officinalis, 4-10 parts of akebia, 4-8 parts of wax gourd peel, 4-8 parts of dried tangerine peel, 2-6 parts of cypress seed, 4-8 parts of Achyranthes bidentata, 2-6 parts of antler glue, 2-6 parts of tortoise shell glue, 6-12 parts of pyrrosia, 1-4 parts of licorice, 4-8 parts of Morinda officinalis, 6-12 parts of mulberry silkworm cocoon, 4-8 parts of Alpinia oxyphylla, 8-16 parts of Rosa laevigata, 8-16 parts of raspberry, and 8-16 parts of cat's whiskers.

[0010] Furthermore, the Guben Yanling Pills for improving frequent nocturnal urination are prepared from the following Chinese medicinal raw materials, calculated by weight: 6 parts of ginseng, 10 parts of yam, 8 parts of cornus officinalis, 8 parts of akebia, 6 parts of wax gourd peel, 6 parts of dried tangerine peel, 4 parts of cypress seed, 6 parts of Achyranthes bidentata, 4 parts of antler glue, 4 parts of tortoise shell glue, 9 parts of pyrrosia, 3 parts of licorice, 6 parts of Morinda officinalis, 9 parts of mulberry silkworm cocoon, 6 parts of Alpinia oxyphylla, 10 parts of Rosa laevigata, 10 parts of raspberry, and 10 parts of cat's whiskers.

[0011] Ginseng: It tastes sweet and slightly bitter, is slightly warm in nature, and enters the spleen, lung, heart, and kidney meridians. It has the effects of replenishing vital energy, restoring the pulse and strengthening the body, nourishing the spleen and lungs, promoting the production of body fluids and nourishing blood, and calming the mind and improving intelligence.

[0012] Chinese Yam: sweet, neutral, enters the spleen, lung and kidney meridians; nourishes the spleen and stomach, promotes fluid production and benefits the lungs, nourishes the kidneys and astringes semen; used for spleen deficiency, poor appetite, chronic diarrhea, lung deficiency, wheezing and coughing, kidney deficiency, spermatorrhea, leucorrhea, frequent urination, deficiency heat and thirst; fried Chinese yam with bran nourishes the spleen and strengthens the stomach; used for spleen deficiency, poor appetite, diarrhea, loose stools, and excessive leucorrhea.

[0013] Cornus officinalis: sour, astringent, slightly warm, enters the liver and kidney meridians. It nourishes the liver and kidneys, astringes the essence and strengthens the body. It is used for vertigo, tinnitus, soreness of the waist and knees, impotence, spermatorrhea, enuresis, frequent urination, metrorrhagia, leukorrhea, profuse sweating, and internal heat and thirst.

[0014] Akebia, bitter in taste, cold in nature, has the effects of diuresis, detumescence, clearing away heat and removing fire.

[0015] Winter melon peel: Sweet in flavor, cold in nature, it enters the spleen and small intestine meridians. It has diuretic and detumescent properties and is used to treat edema, dysuria, thirst caused by summer heat, and short, dark urine. It contains polyphenols, pigments, polysaccharides, and dietary fiber, and has multiple pharmacological effects, including antioxidant, hypoglycemic, antihypertensive, and diuretic properties.

[0016] Pilosula dahurica: spicy in taste, slightly warm in nature, enters the spleen, stomach, large intestine and small intestine meridians; has the effects of promoting qi and relieving fullness, promoting water circulation and reducing swelling, and can be used to treat edema, beriberi, dysuria and other symptoms; mainly contains alkaloids, phenols, oils and other chemical components, and has pharmacological activities such as regulating gastrointestinal function and anti-oxidation.

[0017] Raspberry: Sweet, sour, and warm in nature, it enters the liver, kidney, and bladder meridians. Raspberry nourishes the kidneys, strengthens essence, reduces urination, nourishes the liver, and improves eyesight. It is used to treat impotence, premature ejaculation, spermatorrhea, infertility caused by uterine coldness, thin vaginal discharge, frequent urination, enuresis, blurred vision, and premature graying of hair.

[0018] Platycodon grandiflorum seed: Sweet, neutral, enters the heart, kidney, and large intestine meridians. Nourishes the heart and calms the mind, stops sweating, and moistens the intestines. Used for insomnia due to deficiency, palpitations, night sweats due to yin deficiency, and constipation due to dry intestines.

[0019] Achyranthes bidentata: bitter and sour in taste, neutral in nature, enters the liver and kidney meridians, has the effects of removing blood stasis and dredging menstruation, nourishing the liver and kidneys, strengthening tendons and bones, promoting diuresis and relieving stranguria, and guiding blood downward.

[0020] Deer Antler Glue: Sweet, salty, warming, enters the kidney and liver meridians. Warms and nourishes the liver and kidneys, replenishes essence and blood. Used for impotence and spermatorrhea, soreness and coldness of the waist and knees, consumptive emaciation, metrorrhagia, hematuria, and swelling and pain in the genitals.

[0021] Turtle Shell Glue: Salty, sweet, and cooling. Enters the Liver, Kidney, and Heart meridians. Nourishes Yin, nourishes blood, and stops bleeding. Used for hot flashes due to Yin deficiency, bone steaming, night sweats, soreness of the waist and knees, blood deficiency, sallow complexion, and metrorrhagia and leukorrhea.

[0022] Pyrus nobilis: Sweet and bitter in flavor, it enters the lung and bladder meridians, and has diuretic, stranguria-clearing, cough-relieving, and blood-cooling properties. Containing organic acids, flavonoids, and their glycosides, it is used to treat symptoms such as stranguria caused by heat, urinary obstruction, and painful urination. Pyrus nobilis has a significant diuretic effect, increasing urine volume, helping to excrete excess sugar and toxins from the body, and reducing the burden on the kidneys. It also improves symptoms such as polyuria and edema caused by diabetes. Pyrus nobilis's heat-clearing and detoxifying properties help eliminate heat toxins from the body and improve heat-related symptoms in patients with diabetes, such as thirst and excessive drinking.

[0023] Licorice: It is mild in nature and sweet in taste. It enters the heart, spleen, lung and stomach meridians. Efficacy: Tonic; tonifies the spleen and replenishes qi, clears away heat and detoxifies, eliminates phlegm and relieves cough, relieves urgency and relieves pain, and harmonizes various medicines. It is used for spleen and stomach weakness, fatigue, palpitations and shortness of breath, cough with sputum, cramps and pain in the abdomen and limbs, carbuncles, swellings and sores, and relieves the toxicity and potency of drugs.

[0024] Morinda officinalis: sweet and spicy in taste, slightly warm in nature, enters the liver and kidney meridians, has the effects of replenishing kidney yang, strengthening muscles and bones, and removing rheumatism.

[0025] Mulberry silkworm cocoon: It is neutral in nature, sweet in taste, and enters the liver and kidney meridians. It nourishes the kidneys and boosts yang, consolidates essence and reduces urination. It is used to treat impotence, spermatorrhea, spermatorrhea, enuresis, frequent urination, and excessive leucorrhea caused by kidney deficiency and yang deficiency.

[0026] Alpinia oxyphylla: Pungent and slightly warm in nature, it enters the spleen and kidney meridians. It warms the kidneys and boosts yang, consolidates essence and reduces urination, warms the spleen and stops diarrhea, stimulates appetite and controls saliva. It is used to treat deficiency-cold in the lower abdomen, nocturnal emission and enuresis, leukorrhea, frequent urination, abdominal pain, vomiting and diarrhea, and spontaneous salivation.

[0027] Rosa Laevigata: Astringent in nature, neutral in nature, it enters the bladder, kidney, and large intestine meridians. Rosa Laevigata strengthens essence and astringes the intestines, acts as a diuretic and antidiarrheal, and is used to treat spermatorrhea, wheezing and coughing due to lung deficiency, spontaneous sweating and night sweats, metrorrhagia, leukorrhea, frequent urination, and diarrhea due to spleen deficiency.

[0028] Cat's Whiskers: sweet and slightly bitter in nature, enters the liver and kidney meridians, and has the effects of warming, diuresis and stone removal.

[0029] The preparation method of the Guben Yanling Pill for improving nocturnal frequent urination comprises the following steps:

[0030] (1) Weigh the above raw materials in order by weight;

[0031] (2) The Chinese medicinal raw materials except deer antler glue and tortoise shell glue are crushed and mixed, cellulase and water are added for enzymatic extraction, followed by ultrasonic-assisted extraction, and the first filtrate is obtained by filtration; ethanol aqueous solution is added to the precipitate for secondary extraction, and the second filtrate is obtained by filtration; the two filtrates are combined and concentrated to obtain a Chinese medicinal paste;

[0032] (3) extracting and concentrating the deer antler glue and tortoise shell glue with water to obtain a thick paste;

[0033] (4) Mix the Chinese medicine paste and thick paste, add a binder to make Guben Yanling Pills to improve frequent urination at night.

[0034] Preferably, the amount of cellulase added is 0.5-2 wt% of the total mass of the raw material, the enzymatic hydrolysis pH is 3-5, the temperature is 40-60° C., the stirring speed is 200-600 rpm, and the time is 30-60 min.

[0035] Preferably, the ultrasonic extraction conditions are: power 50-150W, frequency 20-60kHz, temperature 60-90°C, and time 1-3h.

[0036] Preferably, the concentration of the ethanol aqueous solution is 40-60 wt %, the extraction temperature is 70-85° C., and the extraction time is 2-4 h.

[0037] Preferably, the extraction conditions in step (3) are: extraction at 70-90° C. for 1-3 times, with each extraction time being 0.5-2 h.

[0038] Preferably, the binder is one of honey, starch or hydroxypropyl methylcellulose, and the added amount is 2-6wt% of the total paste mass.

[0039] In the present invention, ginseng is the main medicine, which can greatly replenish vital energy, benefit the kidney and assist yang, and can enhance the bladder's gasification function, which is the key to restoring the central power of kidney qi; yam and cornus officinalis are the auxiliary ministers, the former can strengthen the spleen and replenish qi, nourish yin and astringe essence, and the latter can nourish the liver and kidney, consolidate essence and reduce urine, and jointly consolidate the foundation of kidney qi and spleen qi; licorice is the guiding medicine, which harmonizes various medicines, relieves the stimulation of cold and cool medicines, and has the effects of moistening the intestines and promoting bowel movements, and preventing warming and tonic medicines from damaging yin due to dryness and heat. This prescription establishes clear pathways for dispelling dampness and relieving stranguria, as well as clearing heat and detoxifying. A dampness-removing herb group, consisting of Akebia, Winter Melon Peel, Pyrola Rhizoma, and Rhizoma Cibotii, is used to treat stranguria. Akebia promotes diuresis and relieves stranguria, Winter Melon Peel promotes diuresis and reduces swelling, Pyrola Rhizoma dispels dampness and clears turbidity, and Rhizoma Cibotii promotes qi and resolves dampness, promoting diuresis and relieving constipation. Their synergistic effect effectively relieves damp-heat stagnation in the lower burner. Misquito Herba is used as a medicinal herb to clear heat and dampness, expel stones and promote diuresis, and gently regulate the urinary tract metabolic environment. Achyranthes Bidentata activates blood circulation and promotes menstruation, directing blood downward, strengthening the meridians of lower burner drugs and promoting the smooth flow of qi, blood, and water. Furthermore, a pair of deer antler glue and tortoise shell glue is used as a medicinal pair. The former warms and tonifies kidney yang, replenishing essence and benefiting marrow, while the latter nourishes kidney yin, suppressing yang and strengthening the foundation. These two herbs, one warming and one cooling, harmonize yin and yang, strengthening the prescription's essence-replenishing and kidney-strengthening properties. To improve core symptoms such as frequent urination at night, the present invention innovatively combines Rosa Laevigata, Rubus Idaeus, and Herba Miscanthus Officinalis as synergistic intervention units: Rosa Laevigata is rich in tannins and triterpenoid saponins, and has a strong function of consolidating kidney qi and stopping nocturnal emission; Raspberry Idaeus contains a variety of flavonoids and organic acid components, which can enhance the regulatory capacity of the central nervous system, prolong the interval between urinations at night, and enhance urinary control; Herba Miscanthus Officinalis is rich in polyphenols and caffeic acid derivatives, and has the effects of clearing heat and detoxifying, and mildly promoting diuresis, stabilizing the urinary environment, and reducing endogenous dampness and heat. The combination of these three drugs not only complements the pathways and synergizes the targets, but also jointly alleviates the mechanism of nocturnal urination through the ternary mechanism of "consolidation-nerve-promoting diuresis", providing the most core differentiated pharmacodynamic module for the present invention.

[0040] Beneficial effects of the present invention:

[0041] 1. The present invention provides Guben Yanling Pills for improving nocturnal urination and a preparation method thereof. By rationally combining multiple Chinese medicinal herbs based on functional grouping and pathway complementarity, combined with segmented extraction and optimized pill-making processes, this method effectively achieves targeted enrichment and synergistic release of active ingredients. This method overcomes technical bottlenecks such as poor absorption, slow onset, and unstable ingredients in traditional pill formulations, significantly improving the bioavailability and compliance of the preparation.

[0042] 2. In terms of constructing a pathway for promoting diuresis and detoxification, the present invention uses Akebia, Wax Gourd Peel, Selaginella, and Cyperus as the core, which respectively play the role of promoting diuresis and relieving stranguria, reducing swelling and promoting qi, removing dampness and turbidity, and forming an effective system for dredging damp-heat in the lower jiao. In the module for reducing urine and consolidating urine, the five medicinal materials of Sangpiao Coix, Alpinia oxyphylla, Rosa rugosa, Rubus idaeus, and Cat's Whiskers are introduced. Among them, Sangpiao Coix and Alpinia oxyphylla can consolidate essence and reduce urine, and enhance the ability of kidney qi to store; Rosa rugosa is rich in tannins and triterpenoid saponins, which can be astringent and consolidate, and mildly promote diuresis; Raspberry flavonoids and organic acid components can activate neural regulatory pathways, enhance urine control and bladder capacity; polyphenols and caffeic acid derivatives in Cat's Whiskers can clear away heat and detoxify, and regulate the damp-heat environment in the lower jiao. The combination of the three medicines together constitutes a synergistic efficacy pathway of consolidating the body, regulating the mind, and promoting diuresis, which significantly improves the specific efficacy of this prescription for frequent urination at night.

[0043] 3. Animal experiments have demonstrated that the present invention has significant advantages in regulating urination rhythm and reducing nocturnal urination frequency. Compared with the model group, the high-dose Guben Yanling Pill group significantly prolonged the urination latency (from 34.38 minutes to 47.88 minutes, P < 0.05), reduced the urine volume-to-weight ratio (from 44.81 mL / kg to 29.86 mL / kg, P < 0.01), and decreased the total excretion rate (from 81.37% to 62.86%, P < 0.05), demonstrating a clear urine-reducing effect and urinary system regulation, demonstrating its effective intervention for nocturnal urination.

[0044] 4. In terms of safety, histological examination of the Guben Yanling Pills prepared by the present invention revealed no inflammatory damage, cell necrosis, or structural disorder in related organs such as the liver, kidney, adrenal gland, and bladder. Pathological sections revealed neatly arranged hepatocytes, intact renal tubular structure, no adrenal congestion or edema, and dense, orderly smooth muscle in the bladder wall, with no mucosal shedding or inflammatory infiltration. This demonstrates that the formulation exhibits good tissue safety and toxicological stability under long-term use. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0046] Figure 1 The urination latency (min), total excretion rate (%), and urine volume to body weight ratio (mL / kg) of each group of rats after taking the Guben Yanling Pills prepared in Example 1 (n=8).

[0047] Figure 2These are the organ indexes (%) of the liver, kidney, adrenal gland, and bladder tissues of rats in each group after taking the Guben Yanling Pills prepared in Example 1 (n=8).

[0048] Figure 3 HE-stained sections of liver tissue (x100) of rats after taking Guben Yanling Pills prepared in Example 1.

[0049] Figure 4 This is a HE-stained section (x200) of renal tissue of rats after taking the Guben Yanling Pills prepared in Example 1.

[0050] Figure 5 HE-stained sections (x100) of adrenal gland tissue of rats after taking Guben Yanling Pills prepared in Example 1.

[0051] Figure 6 This is a HE-stained section (x100) of the bladder tissue of rats after taking the Guben Yanling Pills prepared in Example 1. DETAILED DESCRIPTION

[0052] The above content of the present invention will be further described in detail below in conjunction with specific embodiments, but it should not be understood that the scope of the above subject matter of the present invention is limited to the following embodiments.

[0053] Some of the raw materials described in this application are commercially available. Other raw materials not described are commercially available:

[0054] Acid cellulase was purchased from Hebei Jiuyu Biotechnology Co., Ltd. with an enzyme activity of 100,000 u / g.

[0055] Starch was purchased from Wuhan Baitong Biotechnology Co., Ltd. and was food grade.

[0056] Hydroxypropyl methylcellulose was purchased from Shenzhen Lefu Biotechnology Co., Ltd. and was food grade.

[0057] Example 1

[0058] A Guben Yanling Pill for improving nocturnal urination is prepared from the following Chinese medicinal raw materials, calculated by weight: 6 parts of ginseng, 10 parts of yam, 8 parts of cornus officinalis, 8 parts of akebia, 6 parts of wax gourd peel, 6 parts of dried tangerine peel, 4 parts of cypress seed, 6 parts of achyranthes bidentata, 4 parts of antler glue, 4 parts of tortoise shell glue, 9 parts of pyrrosia, 3 parts of liquorice, 6 parts of morinda officinalis, 9 parts of silkworm cocoon, 6 parts of alpinia oxyphylla, 10 parts of rose hip, 10 parts of raspberry, and 10 parts of cat's whiskers.

[0059] The preparation method of the Guben Yanling Pill for improving nocturnal frequent urination comprises the following steps:

[0060] (1) Weigh the above raw materials in order by weight;

[0061] (2) Ginseng, Chinese yam, cornus officinalis, akebia, wax gourd peel, dried tangerine peel, cypress seed, Achyranthes bidentata, pyrrosia sinensis, licorice, Morinda officinalis, mulberry silkworm cocoon, Alpinia oxyphylla, Rosa laevigata, raspberry, and cat's whiskers are crushed and mixed, 1 wt% acid cellulase and 8 times the volume of water are added, and heated and stirred at pH = 4, 50°C, and 400 rpm for 40 minutes; then placed under ultrasonic extraction at 80°C, ultrasonic power 100W, and ultrasonic frequency 40kHz for 2 hours, filtered, and obtained a filtrate; 15 times the volume of 50 wt% ethanol aqueous solution is added to the precipitate, extracted at 78°C for 2.5 hours, filtered, and obtained a filtrate; the two filtrates are combined, concentrated under reduced pressure to a relative density of 1.2, and a Chinese medicine paste is obtained for use;

[0062] (3) Mix deer antler glue and tortoise shell glue, add 10 times the volume of water, extract twice at 80℃, each time for 1 hour, combine the two filtrates, and concentrate under reduced pressure to a relative density of 1.2 to obtain a thick paste;

[0063] (4) The above-mentioned Chinese medicine paste and thick paste are mixed to form an ointment, 4 wt% honey is added, and the mixture is made into pills, which are dried to obtain Guben Yanling Pills for improving nocturnal urination.

[0064] Example 2

[0065] A Guben Yanling Pill for improving nocturnal urination is prepared from the following Chinese medicinal ingredients, calculated by weight: 3 parts of ginseng, 15 parts of yam, 3 parts of cornus officinalis, 14 parts of akebia, 3 parts of wax gourd peel, 10 parts of dried tangerine peel, 1 part of cypress seed, 12 parts of achyranthes bidentata, 1 part of antler glue, 7 parts of tortoise shell glue, 5 parts of pyrrosia, 5 parts of liquorice, 3 parts of morinda officinalis, 15 parts of mulberry silkworm cocoon, 3 parts of alpinia oxyphylla, 20 parts of rose hip, 6 parts of raspberry, and 20 parts of cat's whiskers.

[0066] The preparation method of the Guben Yanling Pill for improving nocturnal frequent urination comprises the following steps:

[0067] (1) Weigh the above raw materials in order by weight;

[0068] (2) Ginseng, Chinese yam, cornus officinalis, akebia, wax gourd peel, dried tangerine peel, cypress seed, Achyranthes bidentata, pyrrosia sinensis, licorice, Morinda officinalis, mulberry silkworm cocoon, Alpinia oxyphylla, Rosa laevigata, raspberry, and cat's whiskers are crushed and mixed, 0.5 wt% acid cellulase and 8 times the volume of water are added, and heated and stirred at pH = 5, 40°C, and 600 rpm for 30 minutes; then placed in 90°C, ultrasonic power 50W, and ultrasonic frequency 20kHz for 3 hours, filtered, and obtained a filtrate; 15 times the volume of 40 wt% ethanol aqueous solution is added to the precipitate, extracted at 85°C for 2 hours, filtered, and obtained a filtrate; the two filtrates are combined, concentrated under reduced pressure to a relative density of 1.2, and a Chinese medicine paste is obtained for use;

[0069] (3) Mix deer antler glue and tortoise shell glue, add 10 times the volume of water, extract at 70℃ for 3 times, each time for 0.5h, combine the filtrates from the 2 times, and concentrate under reduced pressure to a relative density of 1.2 to obtain a thick paste;

[0070] (4) The above-mentioned Chinese medicine paste and thick paste are mixed to form an ointment, 2 wt% honey is added, and the mixture is made into pills, which are dried to obtain Guben Yanling Pills for improving nocturnal urination.

[0071] Example 3

[0072] A Guben Yanling Pill for improving nocturnal urination is prepared from the following Chinese medicinal ingredients, calculated by weight: 12 parts of ginseng, 5 parts of yam, 12 parts of cornus officinalis, 2 parts of akebia, 10 parts of wax gourd peel, 2 parts of dried tangerine peel, 8 parts of cypress seed, 3 parts of achyranthes bidentata, 7 parts of antler glue, 1 part of tortoise shell glue, 16 parts of pyrrosia, 0.5 part of liquorice, 10 parts of morinda officinalis, 4 parts of mulberry silkworm cocoon, 10 parts of alpinia oxyphylla, 20 parts of rose hip, 20 parts of raspberry, and 6 parts of cat's whiskers.

[0073] The preparation method of the Guben Yanling Pill for improving nocturnal frequent urination comprises the following steps:

[0074] (1) Weigh the above raw materials in order by weight;

[0075] (2) Ginseng, Chinese yam, cornus officinalis, akebia, wax gourd peel, dried tangerine peel, cypress seed, Achyranthes bidentata, pyrrosia sinensis, licorice, Morinda officinalis, mulberry silkworm cocoon, Alpinia oxyphylla, Rosa laevigata, raspberry, and cat's whisker grass were crushed and mixed, 2 wt% of acid cellulase and 8 times of water were added, and heated and stirred at pH = 3, 60 ° C, and 200 rpm for 60 min; then placed in 60 ° C, ultrasonic power 150 W, ultrasonic frequency 60 kHz for 1 h, filtered to obtain a filtrate; 15 times of the volume of 60 wt% ethanol aqueous solution was added to the precipitate, extracted at 70 ° C for 4 h, filtered to obtain a filtrate; the two filtrates were combined, concentrated under reduced pressure to a relative density of 1.2, and a Chinese medicine paste was obtained for use;

[0076] (3) Mix deer antler glue and tortoise shell glue, add 10 times the volume of water, extract once at 90℃ for 2 hours each time, combine the two filtrates, and concentrate under reduced pressure to a relative density of 1.2 to obtain a thick paste;

[0077] (4) The above-mentioned Chinese medicine paste and thick paste are mixed to form an ointment, 6 wt% starch is added, and the mixture is made into pills, which are dried to obtain Guben Yanling Pills for improving nocturnal urination.

[0078] Example 4

[0079] A Guben Yanling Pill for improving nocturnal urination is prepared from the following Chinese medicinal ingredients, calculated by weight: 4 parts of ginseng, 12 parts of yam, 4 parts of cornus officinalis, 10 parts of akebia, 4 parts of wax gourd peel, 8 parts of dried tangerine peel, 2 parts of cypress seed, 8 parts of achyranthes bidentata, 2 parts of antler glue, 6 parts of tortoise shell glue, 6 parts of pyrrosia, 4 parts of liquorice, 4 parts of morinda officinalis, 12 parts of silkworm cocoon, 4 parts of alpinia oxyphylla, 16 parts of rose hip, 8 parts of raspberry, and 8 parts of cat's whiskers.

[0080] The preparation method of the Guben Yanling Pill for improving nocturnal frequent urination comprises the following steps:

[0081] (1) Weigh the above raw materials in order by weight;

[0082] (2) Ginseng, Chinese yam, cornus officinalis, akebia, wax gourd peel, dried tangerine peel, cypress seed, Achyranthes bidentata, pyrrosia sinensis, licorice, Morinda officinalis, mulberry silkworm cocoon, Alpinia oxyphylla, Rosa laevigata, raspberry, and cat's whisker grass were crushed and mixed, 1.5 wt% acid cellulase and 8 times the volume of water were added, and heated and stirred at pH = 4.5, 55°C, and 300 rpm for 45 minutes; then placed under ultrasonic extraction at 70°C, ultrasonic power 80W, and ultrasonic frequency 30kHz for 1.5 hours, filtered, and obtained a filtrate; 15 times the volume of 55 wt% ethanol aqueous solution was added to the precipitate, extracted at 75°C for 3 hours, filtered, and obtained a filtrate; the two filtrates were combined, concentrated under reduced pressure to a relative density of 1.2, and a Chinese medicine paste was obtained for use;

[0083] (3) Mix deer antler glue and tortoise shell glue, add 10 times the volume of water, extract twice at 75℃, each time for 1.5 hours, combine the two filtrates, and concentrate under reduced pressure to a relative density of 1.2 to obtain a thick paste;

[0084] (4) The above-mentioned Chinese medicine paste and thick paste are mixed to form an ointment, 4 wt% honey is added, and the mixture is made into pills, which are dried to obtain Guben Yanling Pills for improving nocturnal urination.

[0085] Example 5

[0086] A Guben Yanling Pill for improving nocturnal urination is prepared from the following Chinese medicinal ingredients, calculated by weight: 8 parts of ginseng, 8 parts of yam, 10 parts of cornus officinalis, 4 parts of akebia, 8 parts of wax gourd peel, 4 parts of dried tangerine peel, 6 parts of cypress seed, 4 parts of achyranthes bidentata, 6 parts of antler glue, 2 parts of tortoise shell glue, 12 parts of pyrrosia, 1 part of liquorice, 8 parts of morinda officinalis, 6 parts of silkworm cocoon, 8 parts of alkyl oxyphylla, 8 parts of rosa laevigata fruit, 16 parts of raspberry, and 16 parts of cat's whiskers.

[0087] The preparation method of the Guben Yanling Pill for improving nocturnal frequent urination comprises the following steps:

[0088] (1) Weigh the above raw materials in order by weight;

[0089] (2) Ginseng, Chinese yam, cornus officinalis, akebia, wax gourd peel, dried tangerine peel, cypress seed, Achyranthes bidentata, stone vine, licorice, Morinda officinalis, mulberry silkworm cocoon, Alpinia oxyphylla, Rosa laevigata, raspberry, and cat's whisker grass were crushed and mixed, 1.8 wt% of acid cellulase and 8 times the volume of water were added, and heated and stirred at pH = 3.5, 45 ° C, and 500 rpm for 50 min; then placed in 85 ° C, ultrasonic power 120 W, ultrasonic frequency 50 kHz for 2.5 h, filtered to obtain a filtrate; 15 times the volume of 45 wt% ethanol aqueous solution was added to the precipitate, extracted at 80 ° C for 3.5 h, filtered to obtain a filtrate; the two filtrates were combined, concentrated under reduced pressure to a relative density of 1.2, and a Chinese medicine paste was obtained for use;

[0090] (3) Mix deer antler glue and tortoise shell glue, add 10 times the volume of water, extract twice at 85℃, each time for 1 hour, combine the two filtrates, and concentrate under reduced pressure to a relative density of 1.2 to obtain a thick paste;

[0091] (4) The above-mentioned Chinese medicine paste and thick paste are mixed to form an ointment, 5 wt% hydroxypropyl methylcellulose is added, and the mixture is made into pills, which are dried to obtain Guben Yanling Pills for improving nocturnal urination.

[0092] Comparative Example 1

[0093] The invention is basically the same as Example 1, except that: a Guben Yanling Pill for improving frequent urination at night is prepared from the following Chinese medicinal raw materials, calculated by weight: 6 parts of ginseng, 10 parts of yam, 8 parts of cornus officinalis, 8 parts of akebia, 6 parts of wax gourd peel, 6 parts of dried tangerine peel, 4 parts of cypress seed, 6 parts of Achyranthes bidentata, 4 parts of antler glue, 4 parts of tortoise shell glue, 9 parts of pyrrosia, 3 parts of licorice, and 6 parts of Morinda officinalis.

[0094] Comparative Example 2

[0095] The invention is basically the same as Example 1, except that: a Guben Yanling Pill for improving frequent urination at night is prepared from the following Chinese medicinal raw materials, calculated by weight: 6 parts of ginseng, 10 parts of yam, 8 parts of cornus officinalis, 4 parts of cypress seed, 6 parts of Achyranthes bidentata, 4 parts of antler glue, 4 parts of tortoise shell glue, 3 parts of licorice, 6 parts of Morinda officinalis, 9 parts of mulberry silkworm cocoon, 6 parts of Alpinia oxyphylla, 10 parts of Rosa laevigata, 10 parts of raspberry, and 10 parts of cat's whiskers.

[0096] Comparative Example 3

[0097] The invention is basically the same as Example 1, except that: a Guben Yanling Pill for improving frequent urination at night is prepared from the following Chinese medicinal raw materials, calculated by weight: 6 parts of ginseng, 10 parts of yam, 8 parts of cornus officinalis, 8 parts of akebia, 6 parts of wax gourd peel, 6 parts of dried tangerine peel, 4 parts of cypress seed, 6 parts of Achyranthes bidentata, 9 parts of Pyrola, 3 parts of licorice, 6 parts of Morinda officinalis, 9 parts of mulberry silkworm cocoon, 6 parts of Alpinia oxyphylla, 10 parts of Rosa laevigata, 10 parts of Rubus idaeus, and 10 parts of Cat's Whiskers.

[0098] Comparative Example 4

[0099] The invention is basically the same as Example 1, except that: a Guben Yanling Pill for improving frequent urination at night is prepared from the following Chinese medicinal raw materials, calculated by weight: 6 parts of ginseng, 10 parts of yam, 8 parts of cornus officinalis, 8 parts of akebia, 6 parts of wax gourd peel, 6 parts of dried tangerine peel, 6 parts of Achyranthes bidentata, 4 parts of antler glue, 4 parts of tortoise shell glue, 9 parts of pyrrosia, 3 parts of licorice, 6 parts of Morinda officinalis, 9 parts of mulberry silkworm cocoon, 6 parts of Alpinia oxyphylla, 10 parts of Rosa laevigata, 10 parts of raspberry, and 10 parts of cat's whiskers.

[0100] Comparative Example 5

[0101] The invention is basically the same as Example 1, except that: a Guben Yanling Pill for improving frequent urination at night is prepared from the following Chinese medicinal raw materials, calculated by weight: 6 parts of ginseng, 10 parts of yam, 8 parts of cornus officinalis, 8 parts of akebia, 6 parts of wax gourd peel, 6 parts of dried tangerine peel, 4 parts of cypress seed, 6 parts of Achyranthes bidentata, 4 parts of antler glue, 4 parts of tortoise shell glue, 9 parts of pyrrosia, 3 parts of licorice, 6 parts of Morinda officinalis, 9 parts of mulberry silkworm cocoon, 6 parts of Alpinia oxyphylla, 15 parts of raspberry, and 15 parts of cat's whiskers.

[0102] Comparative Example 6

[0103] The invention is basically the same as Example 1, except that: a Guben Yanling Pill for improving frequent urination at night is prepared from the following Chinese medicinal raw materials, calculated by weight: 6 parts of ginseng, 10 parts of yam, 8 parts of cornus officinalis, 8 parts of akebia, 6 parts of wax gourd peel, 6 parts of dried tangerine peel, 4 parts of cypress seed, 6 parts of Achyranthes bidentata, 4 parts of antler glue, 4 parts of tortoise shell glue, 9 parts of pyrrosia, 3 parts of licorice, 6 parts of Morinda officinalis, 9 parts of mulberry silkworm cocoon, 6 parts of Alpinia oxyphylla, 15 parts of Rosa laevigata, and 15 parts of raspberry.

[0104] Comparative Example 7

[0105] The invention is basically the same as Example 1, except that: a Guben Yanling Pill for improving frequent urination at night is prepared from the following Chinese medicinal raw materials, calculated by weight: 6 parts of ginseng, 10 parts of yam, 8 parts of cornus officinalis, 8 parts of akebia, 6 parts of wax gourd peel, 6 parts of dried tangerine peel, 4 parts of cypress seed, 6 parts of Achyranthes bidentata, 4 parts of antler glue, 4 parts of tortoise shell glue, 9 parts of pyrrosia, 3 parts of licorice, 6 parts of Morinda officinalis, 9 parts of mulberry silkworm cocoon, 6 parts of Alpinia oxyphylla, 15 parts of Rosa laevigata, and 15 parts of cat's whiskers.

[0106] Comparative Example 8

[0107] The invention is basically the same as Example 1, except that: a Guben Yanling Pill for improving frequent urination at night is prepared from the following Chinese medicinal raw materials, calculated by weight: 6 parts of ginseng, 10 parts of yam, 8 parts of cornus officinalis, 8 parts of akebia, 6 parts of wax gourd peel, 6 parts of dried tangerine peel, 4 parts of cypress seed, 6 parts of Achyranthes bidentata, 4 parts of antler glue, 4 parts of tortoise shell glue, 9 parts of pyrrosia, 3 parts of licorice, 6 parts of Morinda officinalis, 9 parts of mulberry silkworm cocoons, 6 parts of Alpinia oxyphylla, and 30 parts of Rosa laevigata.

[0108] Comparative Example 9

[0109] The invention is basically the same as Example 1, except that: a Guben Yanling Pill for improving frequent urination at night is prepared from the following Chinese medicinal raw materials, calculated by weight: 6 parts of ginseng, 10 parts of yam, 8 parts of cornus officinalis, 8 parts of akebia, 6 parts of wax gourd peel, 6 parts of dried tangerine peel, 4 parts of cypress seed, 6 parts of Achyranthes bidentata, 4 parts of antler glue, 4 parts of tortoise shell glue, 9 parts of pyrrosia, 3 parts of licorice, 6 parts of Morinda officinalis, 9 parts of mulberry silkworm cocoon, 6 parts of Alpinia oxyphylla, and 30 parts of raspberry.

[0110] Comparative Example 10

[0111] The invention is basically the same as Example 1, except that: a Guben Yanling Pill for improving frequent urination at night is prepared from the following Chinese medicinal raw materials, calculated by weight: 6 parts of ginseng, 10 parts of yam, 8 parts of cornus officinalis, 8 parts of akebia, 6 parts of wax gourd peel, 6 parts of dried tangerine peel, 4 parts of cypress seed, 6 parts of Achyranthes bidentata, 4 parts of antler glue, 4 parts of tortoise shell glue, 9 parts of pyrrosia, 3 parts of licorice, 6 parts of Morinda officinalis, 9 parts of mulberry silkworm cocoon, 6 parts of Alpinia oxyphylla, and 30 parts of cat's whiskers.

[0112] Comparative Example 11

[0113] The invention is basically the same as Example 1, except that: a Guben Yanling Pill for improving frequent urination at night is prepared from the following Chinese medicinal raw materials, calculated by weight: 6 parts of ginseng, 10 parts of yam, 8 parts of cornus officinalis, 8 parts of akebia, 6 parts of wax gourd peel, 6 parts of dried tangerine peel, 4 parts of cypress seed, 6 parts of Achyranthes bidentata, 4 parts of antler glue, 4 parts of tortoise shell glue, 9 parts of pyrrosia, 3 parts of licorice, 6 parts of Morinda officinalis, 9 parts of mulberry silkworm cocoons, and 6 parts of Alpinia oxyphylla.

[0114] Comparative Example 12

[0115] It is basically the same as Example 1, except that:

[0116] The preparation method of the Guben Yanling Pill for improving nocturnal frequent urination comprises the following steps:

[0117] (1) Weigh the above raw materials in order by weight;

[0118] (2) Ginseng, Chinese yam, cornus officinalis, akebia, wax gourd peel, dried tangerine peel, cypress seed, Achyranthes bidentata, pyrrosia sinensis, licorice, Morinda officinalis, mulberry silkworm cocoon, Alpinia oxyphylla, Rosa laevigata, raspberry, cat's whisker grass, deer antler glue and tortoise shell glue were crushed and mixed, 1 wt% acid cellulase and 8 times the volume of water were added, and heated and stirred at pH = 4, 50 ° C, and 400 rpm for 40 minutes; then placed under ultrasonic extraction at 80 ° C, ultrasonic power 100 W, and ultrasonic frequency 40 kHz for 2 hours, filtered to obtain a filtrate; 15 times the volume of 50 wt% ethanol aqueous solution was added to the precipitate, extracted at 78 ° C for 2.5 hours, filtered to obtain a filtrate; the two filtrates were combined, concentrated under reduced pressure to a relative density of 1.2, and a Chinese medicine paste was obtained for use;

[0119] (3) The above-mentioned Chinese medicine paste is made into an ointment, 4 wt% honey is added, and the mixture is made into pills, which are dried to obtain Guben Yanling Pills for improving nocturnal urination.

[0120] Comparative Example 13

[0121] Basically the same as Example 1, 60g of ginseng, 60g of ophiopogon, 60g of schisandra, 60g of asparagus, 60g of rehmannia, 60g of prepared rehmannia, 60g of yam, 30g of oriental rhizome, 60g of poria, 60g of cyperus, 60g of cornus officinalis, 60g of eucommia, 60g of costus root, 30g of polygala, 60g of cypress seed, 60g of morinda officinalis (salted), 30g of zanthoxylum bungeanum, 30g of calamus, 120g of salvia miltiorrhiza, 120g of cistanche deserticola, 45g of wolfberry, 45g of raspberry, 45g of rehmannia root, 30g of antler glue, 120g of dodder seed, 30g of fish maw beads, 9g of pearl, and 25g of dog whip.

[0122] The preparation method is basically the same as that in Example 1.

[0123] Test Example 1

[0124] DPPH free radical scavenging ability determination: 0.1 mL of the 1 mg / mL methanol solution of Guben Yanling Pills in the above examples and comparative examples was added to 3.9 mL 2 .5×10 -2 The mixture was added into 100 mg / mL DPPH methanol solution, and the mixture was incubated in a dark place at room temperature for 30 min. The absorbance was measured at 517 nm.

[0125] The percentage scavenging rate of DPPH free radicals was calculated according to the formula (1): (1): DPPH free radical scavenging rate = (A0-A) / A0×100%, where: A0-absorbance after the reaction without adding the test solution; A-absorbance measured after the reaction with the test solution. The DPPH free radical scavenging rate of Guben Yanling Pills for improving nocturnal urination was determined. Each group was tested four times and the average value was taken. The results are shown in Table 1.

[0126] Table 1 Free radical scavenging rate and urine reduction test results of Guben Yanling Pills for improving frequent urination at night

[0127] DPPH free radical scavenging rate, % Urinary latency, min Total excretion, % Example 1 89.6 47.88 62.86 Example 2 85.5 45.23 65.40 Example 3 86.1 46.50 64.36 Example 4 87.4 46.80 63.70 Example 5 87.9 47.22 63.23 Comparative Example 1 80.7 38.50 72.55 Comparative Example 2 82.4 40.85 70.20 Comparative Example 3 83.0 42.10 68.96 Comparative Example 4 78.6 36.70 75.30 Comparative Example 5 85.6 44.67 66.80 Comparative Example 6 86.4 45.94 65.58 Comparative Example 7 84.3 43.26 67.45 Comparative Example 8 82.0 41.50 69.70 Comparative Example 9 82.9 44.00 66.23 Comparative Example 10 81.8 39.83 71.50 Comparative Example 11 79.2 37.20 73.86 Comparative Example 12 80.7 39.54 72.36 Comparative Example 13 83.4 30.42 78.94

[0128] As can be seen from the above experimental results, the Guben Yanling Pills prepared by the present invention have significant antioxidant activity and urine reduction effect. Specifically, in Examples 1 to 5, the DPPH free radical scavenging rate and urination latency were significantly increased, while the total urine volume was reduced, which was better than the overall combination of each comparative example. Its advantage is that through the compound system design, the multi-target synergistic effect between the medicinal materials is fully utilized. In the overall compatibility, ginseng, yam, and licorice are the tonic medicine group, rich in saponins and polysaccharide components, which directly scavenge free radicals and enhance basal metabolism and antioxidant capacity; Akebia, wax gourd peel, Pyrola, and Piper melongena are the most diuretic medicine group, rich in polyphenols, flavonoids and organic acids, which synergistically promote the scavenging of metabolic waste and free radicals and strengthen the antioxidant chain in the body. In addition, the most distinctive design of the present invention is the synergistic compatibility of Rosa laevigata, Rubus idaeus and Miscanthus sinensis. Although these three medicinal materials have their own specialties in traditional astringent drugs, they form a synergistic effect of urine reduction, regulating the mind, and promoting dampness, forming a multi-target, multi-pathway intervention mechanism. Rosa laevigata fruit, rich in tannins and triterpenoid saponins, helps strengthen kidney qi and reduce involuntary urination. The flavonoids in raspberry can enhance central nervous system inhibition, increasing bladder capacity and urinary control threshold at night. And cat's whiskers, with its caffeic acid derivatives and polyphenols, offer mild diuretic properties and reduce bladder irritation. Combined use of these three drugs can improve nocturia while effectively modulating neural reflex pathways and the local bladder environment, achieving comprehensive intervention across multiple levels, encompassing the causes, pathogenesis, and symptoms.

[0129] As can be seen from the experimental results of Example 1 and Comparative Examples 5-11, Example 1 achieved a DPPH free radical scavenging rate of 89.6%, a prolonged urination latency of 47.88 minutes, and a reduced total excretion volume of 62.86%, significantly outperforming the corresponding data in each comparative example (DPPH scavenging rate of 79.2% to 86.4%, urination latency of 37.2 to 45.94 minutes, and total excretion volume of 65.58% to 73.86%). This demonstrates that the synergistic combination of the core medicinal ingredients of Rosa laevigata, Rubus idaeus, and Herba Miscanthi in Example 1, targeting the formation mechanism of nocturia, establishes a multi-target intervention system with strengthening the foundation, regulating the mind, and promoting diuresis as the main themes. The combined use of equal amounts of these three medicinal ingredients, as verified by in vitro antioxidant experiments and animal models, was superior to the comparative example groups composed of any single or two medicinal ingredients, demonstrating clear functional complementarity and synergistic efficacy. This synergistic mechanism not only significantly enhances overall antioxidant capacity but also strengthens the regulatory stability of the urinary system, achieving a balanced and optimized overall internal environment while improving nocturia symptoms. Among them, Rosa Laevigatae, rich in tannins and triterpenoid saponins, strengthens bladder sphincter tone and strengthens kidney qi. Iberis idaeus, with its flavonoids and organic acids, helps regulate central nervous system excitability and prolong the latency of the urinary control reflex. And Misquito Herba, with its polyphenols and caffeic acid derivatives, clears heat and dampness, improving the environment of the bladder and lower urethra. These three ingredients act on different links in the etiology, forming a stable closed loop of strengthening the foundation, regulating the mind, and promoting dampness removal, thus achieving the synergistic and effective compound advantage.

[0130] It can be seen from Example 1 and Comparative Example 13 that the improvement of nocturnal urination prepared by the present invention is significantly better than that of Comparative Example 13. The reason is that Example 1 forms a synergistic mechanism of consolidating, regulating the mind and promoting diuresis through the precise combination of core medicinal materials such as Rosa laevigata, Rubus idaeus and Herba Miscanthuse, which specifically improves nocturnal urination; while Comparative Example 13 (National Standard Guben Yanling Pills) focuses on nourishing and prolonging life, lacks a specific disease-targeted design, and therefore has a weaker effect on reducing urine output.

[0131] 2. Experimental Animals

[0132] Male Sprague-Dawley rats weighing (220 ± 20) g were purchased from the Animal Science and Technology Center of Jiangxi University of Traditional Chinese Medicine under license number SCXK (Gan) 2023-0001. The animals were housed at a temperature of 25.0 ± 1.5°C, a relative humidity of 45 ± 15%, and a 12 / 12-hour light / dark cycle. Experiments were conducted after one week of adaptive feeding. Animal welfare regulations and the 3R principles were adhered to.

[0133] 2.1 Experimental methods

[0134] Water load polyuria model experiment:

[0135] Preparation of the medicine: Take an appropriate amount of Guben Yanling Pills, cut them into pieces, put them in a mortar and pestle, add physiological saline and grind them into a uniform medicinal solution. Then prepare three concentrations of 0.6 g / mL, 0.3 g / mL and 0.15 g / mL respectively and store them for later use.

[0136] Suquan pills (purchased from Guangzhou Shaxi Pharmaceutical Co., Ltd.; batch number: 2311010): Grind an appropriate amount of Suquan pills, place them in a mortar, add 0.5% sodium carboxymethyl cellulose and grind them into a uniform liquid, then prepare a liquid with a concentration of 0.133 g / mL and store it for later use.

[0137] 2.2 Animal screening

[0138] SD rats were fasted for 18 hours before the experiment but not water. Animals were screened using the Aston method, with 22 mL / kg of deionized water perfused according to body weight to achieve water balance. Urine was collected over a 2-hour period; rats were considered qualified if their urine volume exceeded 40% of their water load.

[0139] 2.3 Grouping and Dosing

[0140] Forty-eight rats that passed the screening were evenly divided into six groups based on urine volume and body weight: a normal group, a model group, a Suquan Wan group, and high-, medium-, and low-dose groups of Guben Yanling Wan prepared in Example 1, with eight animals in each group. The Suquan Wan dose was 2 g / kg, while the high, medium, and low doses of Guben Yanling Wan were 9, 4.5, and 2.25 g / kg, respectively. The animals in the normal group served as a control reference for the mental state, behavioral activity, and urination status of the animals in each group.

[0141] Each group received 15 mL / kg of the drug by gavage for one week. The normal and model groups received an equal volume of distilled water, and the remaining groups received the corresponding drug solution. Animals were fasted for 18 hours before the final dose. One hour after the drug administration, they were gavaged with 50 mL / kg of purified water. The normal group received no water. The bladders were squeezed to empty, and the rats in each group were placed in metabolic cages and their urine output and time to first urination were observed for 6 hours.

[0142] 2.4 Observation and testing of indicators

[0143] 2.4.1 General appearance

[0144] Compared with the animals in the normal group, the daily mental state, behavioral activities, eating and drinking conditions, hair gloss, feces and urine of the animals in the model group and each drug-treated group were improved.

[0145] 2.4.2 Urine reduction effect index

[0146] The urination latency was determined based on the time of first urination after water loading. The total excretion rate of each group of animals was calculated based on the total urine volume within 6 hours. The urine volume to body weight ratio was calculated based on the ratio of 6-hour urine volume to body weight. The specific formula is as follows: Total excretion rate (%) = 6-hour total urine volume / water loading amount × 100%

[0147] Urine volume to body weight ratio mL / kg = rat urine volume mL / rat weight kg

[0148] 2.4.3 Organ indices of liver, kidney, adrenal gland, and bladder

[0149] At the end of the experiment, the animals were sacrificed, the abdominal and thoracic cavities were opened, and the liver, kidneys, adrenal glands, and bladder tissues were carefully removed. The corresponding tissues were removed according to a unified technique, washed with saline, and weighed after absorbing water with filter paper. Organ index = organ weight / body weight × 100%

[0150] 2.4.4 Histopathological examination

[0151] The tissues retained in the experiment were stained with HE (hematoxylin-eosin) for histopathological examination.

[0152] The specific operation method and process take liver tissue as an example: (The specific parameters of the different tissue processing processes are determined based on preliminary experiments)

[0153] Tissue embedding and sectioning

[0154] Trimming: Each group of liver tissue was removed from the fixative and trimmed in a fume hood. The tissue was then rinsed in running water overnight and then dehydrated.

[0155] Dehydration: Place the rinsed tissue in 75% alcohol for 4 hours, 85% alcohol for 2 hours, 90% alcohol for 1.5 hours, 95% alcohol for 1 hour, and anhydrous ethanol I and II for 30 minutes each.

[0156] Transparency: Immerse the dehydrated tissue in xylene: anhydrous ethanol (1:1) for 10 minutes, and xylene I and II for 20 minutes each.

[0157] Wax immersion: Immerse the transparent tissue in 60℃ paraffin I, II, and III for 1 hour each.

[0158] Embedding: Pour liquid paraffin into the embedding base mold, then place the fully infiltrated tissue block flat on the bottom, with the cut surface facing downwards, and then transfer it as a whole to a cold table to cool. After the paraffin is fully solidified, remove the embedding base mold, trim the wax block and store it at room temperature for subsequent sectioning.

[0159] Spreading the slices: Adjust the angles of the slice and the blade to make them parallel, then fix the tissue paraffin block in the slice slot. Hold the brush in your left hand and turn the microtome handle with your right hand. After cutting, hold the slices with the brush. The slices are 4μm thick. Then, gently pick them up with tweezers and place them in a slide spreader for spreading. The water temperature is 40℃. After the slices are completely spread, hold a glass slide and vertically immerse it in water to attach the slices. Then, bake the slides in a 60℃ oven for 30 minutes.

[0160] HE staining process

[0161] Dewax to water: Bake the paraffin sections in a 60°C oven for 1 hour, then quickly dewax them in xylene I and II for 5 minutes. Repeat this operation twice. Then hydrate the sections and immerse them in anhydrous ethanol I and II for 5 minutes each, 95% alcohol for 5 minutes, 85% alcohol for 5 minutes, and 75% alcohol for 5 minutes. Then rinse them with tap water and distilled water several times, each for about 5 minutes.

[0162] Hematoxylin staining of cell nuclei: The hydrated liver tissue sections were immersed in hematoxylin staining solution for 5 minutes, then rinsed with tap water for 2 minutes, then placed in 1% hydrochloric acid alcohol for differentiation for 10 seconds, rinsed with tap water for 10 seconds, then placed in 2% sodium bicarbonate for blueing for 40 seconds, and washed with water for 2 minutes.

[0163] Eosin staining of cytoplasm: Stain the sections in eosin staining solution for 2 minutes, then rinse with running water for 1 minute.

[0164] Dehydration and clearing: Place the sections in 95% alcohol I, 95% alcohol II, anhydrous ethanol I, and anhydrous ethanol II for 5 minutes each, then transfer to xylene I and II for clearing for 5 minutes.

[0165] Seal film and take photos

[0166] The sections were sealed with neutral gum and the changes in tissue morphology were observed under a microscope and photographed for preservation.

[0167] 2.5 Animal Handling and Collection

[0168] After the experiment, rats were fasted for 12 hours but not water. After anesthesia, blood was collected from the abdominal aorta and sacrificed. Blood was collected in centrifuge tubes and allowed to clot at room temperature for 2 hours. Serum was then centrifuged at 3000 rpm for 10 minutes to separate the serum. The serum was aliquoted into 1.5 mL EP tubes and stored at -80°C until needed.

[0169] Carefully open the abdominal and thoracic cavities and carefully dissect the heart, liver, kidneys, adrenal glands, spleen, and bladder. Remove the surface mucosa and adipose tissue, wash away any blood, blot dry with filter paper, and weigh. Samples of tissue from the same location in each group were excised and fixed in 4% paraformaldehyde for histopathological analysis.

[0170] 2.6 Statistical Analysis of Data

[0171] GraphPad Prism statistical software was used for data processing, and the results were expressed as mean ± SD. One-way ANOVA was used for significance analysis. P > 0.05 indicated no statistically significant difference, P < 0.05 indicated statistically significant difference, and P < 0.01 indicated statistically significant difference.

[0172] 3. Experimental results

[0173] 3.1 General appearance

[0174] The animals in each group were in good spirits and behaved freely every day; there was no significant difference in food intake, water intake, hair gloss, feces and urine compared with the animals in the normal group.

[0175] 3.2 Urine reduction effect index

[0176] The time interval from the start of water loading to the first urination of each group of animals is the urination latency period. The urine excretion rate and urine volume to body weight ratio within 6 hours after water loading can reflect the drug's urine reduction effect, that is, improving the physiological phenomenon of frequent urination at night. As shown in Table 2, Figure 1 As shown in the figure (Note: *P<0.05, statistically significant difference; **P<0.01, statistically extremely significant difference) (compared with the water-load polyuria model group (M), each group had a prolonged urination latency period in the Suquan pill group (SQW) and the high- and medium-dose Guben Yanling pill groups compared with the water-load polyuria model group (M), with statistically significant differences (P<0.05), which played a role in delaying urination. The total excretion rate within 6 hours was significantly lower than that in the water-load polyuria model group (M). Compared with the water-loaded polyuria model group (M), the total excretion rate was reduced in the Suquan Pills group (SQW) and the high-, medium-, and low-dose Guben Yanling Pills groups, and the difference was statistically significant (P < 0.05); in terms of the urine volume to body weight ratio, compared with the water-loaded polyuria model group (M), the Suquan Pills group (SQW) and the medium- and low-dose Guben Yanling Pills groups showed significant differences (P < 0.05), while the high-dose Guben Yanling Pills group showed an extremely significant difference (P < 0.01).

[0177] Table 2: Urinary latency (min), total excretion rate (%), and urine volume to body weight ratio (mL / kg) of animals in each group (n=8)

[0178] Group Dosage (g / kg) Urinary latency (min) Total excretion rate (%) Urine volume to body weight ratio (mL / kg) Water load polyuria model group (M) / 34.38±7.63 81.37±9.38 44.81±3.56 Suquanwan group (SQW) 2 <![CDATA[48.88±11.57 ✱ ]]> <![CDATA[58.85±21.39 ✱ ]]> <![CDATA[30.70±11.69 ✱ ]]> Guben Yanling Pills high-dose group (GB-H) 9 <![CDATA[47.88±9.70 ✱ ]]> <![CDATA[62.86±8.10 ✱ ]]> <![CDATA[29.86±9.95 ✱✱ ]]> Guben Yanling Pills medium dose group (GB-M) 4.5 <![CDATA[48.25±10.38 ✱ ]]> <![CDATA[61.88±12.26 ✱ ]]> <![CDATA[32.85±7.55 ✱ ]]> Guben Yanling Pills low-dose group (GB-L) 2.25 43.63±12.01 <![CDATA[61.13±15.56 ✱ ]]> <![CDATA[32.33±10.69 ✱ ]]>

[0179] Note: *P<0.05, statistically significant difference between each group and the water-load polyuria model group (M); **P<0.01, statistically extremely significant difference.

[0180] 3.3 Organ Index

[0181] The organ indices of liver tissue, which is directly affected by drug metabolism, and kidney, adrenal gland, and bladder tissue, which are closely related to urine reduction, were selected to roughly reflect the effects of drugs on tissues and organs. Figure 2 As shown (Note: *P < 0.05, statistically significant difference; **P < 0.01, extremely significant difference; #P < 0.05, statistically significant difference compared with the normal group (N)), there were no statistically significant differences in the liver, kidney, adrenal gland, and bladder organ indices of the animals in the water-load polyuria model group (M) compared with the animals in the normal group (N). Compared with the animals in the normal group (N), the adrenal gland organ indices of the high- and medium-dose Guben Yanling Pill groups were statistically significant (P < 0.05). Compared with the water-load polyuria model group (M), the adrenal gland organ indices of the high-dose Guben Yanling Pill group were extremely significant (P < 0.01).

[0182] These results suggest that the water-loaded polyuria model does not affect the overall status of the liver, kidneys, adrenal glands, or bladder tissues; nor do the various dosing groups affect the overall status of the liver, kidneys, and bladder tissues. The high- and medium-dose Guben Yanling Pills groups showed some effects on adrenal organ indices, possibly related to their ability to regulate urine metabolism.

[0183] Table 3: Organ index indicators (%) of liver, kidney, adrenal gland and bladder tissues of animals in each group (n=8)

[0184] Group liver kidney adrenal glands bladder Normal group (N) 2.81±0.45 0.72±0.04 0.011±0.001 0.027±0.005 Water load polyuria model group (M) 2.69±0.10 0.73±0.04 0.010±0.001 0.026±0.005 Suquanwan group (SQW) 2.79±0.25 0.70±0.05 0.009±0.002 0.024±0.004 Guben Yanling Pills high-dose group (GB-H) 2.57±0.15 0.73±0.08 <![CDATA[0.016±0.006 ✱✱＃ ]]> 0.026±0.004 Guben Yanling Pills medium dose group (GB-M) 2.82±0.24 0.68±0.06 <![CDATA[0.007±0.002 ＃ ]]> 0.023±0.003 Guben Yanling Pills low-dose group (GB-L) 2.54±0.30 0.67±0.05 0.013±0.002 0.029±0.004

[0185] Note: Compared with the water-load polyuria model group (M), *P<0.05, there are statistically significant differences; **P<0.01, there are extremely significant differences; compared with the normal group (N), #P<0.05, there are statistically significant differences.

[0186] 3.4 HE staining results of liver, kidney, adrenal gland, and bladder tissues

[0187] During the animal treatment and sampling process, it was observed that the abdominal cavity, thoracic cavity, heart, liver, spleen, lung, kidney, adrenal gland and bladder of the rats in each group were in good condition and showed no difference compared with the normal group.

[0188] Liver tissue, which is directly affected by drug metabolism, and kidney, adrenal gland, and bladder tissue, which are closely related to urine metabolism and regulation, were selected for HE staining to detect pathological changes in each tissue. Figure 3 、 4 , 5, 6.

[0189] Depend on Figure 3 As shown in the data, in the liver tissue sections of animals in the normal group, the water-loaded polyuria model group, and each drug-administered group, hepatocytes were evenly distributed, with cells arranged tightly and regularly. The hepatocytes were normal in morphology, polygonal in shape, and arranged in a lamellar or radial pattern with the central vein as the center. The nuclei were mostly round, located in the center of the cell, of uniform size and evenly distributed, with clear nuclear membranes, and no abnormal condensation or marginalization of chromatin. The cytoplasm was stained evenly with consistent depth, no obvious vacuolar changes were observed, and no inflammatory cell infiltration was observed. The results showed that the construction process of the water-loaded polyuria model and the treatment of each drug-administered group did not cause obvious pathological damage to the liver tissue.

[0190] like Figure 4 As shown, renal tissue sections from animals in the normal group, the water-load polyuria model group, and each drug-treated group showed regular glomerular morphology, well-patented capillary loops, and no proliferation of mesangial cells or matrix. Periglomerular tubules were neatly arranged, with intact epithelial cell morphology and clear tubular lumens, and no vacuolar degeneration, necrosis, or cast formation was observed. The renal capsule cavity was not narrowed or dilated, and the cyst wall structure was clear. No pathological changes such as inflammatory cell infiltration, vascular congestion, or congestion were observed in the renal tissue of any group. These results indicate that during the establishment of the water-load polyuria model and subsequent treatment in each drug-treated group, no obvious signs of pathological damage were observed in the renal tissue at the cellular and histological levels.

[0191] like Figure 5 As shown, the adrenal tissue cells in the normal group were densely arranged with clear layers, chromatin was evenly distributed, the nuclear membrane was clear, and there was no pyknosis, fragmentation, or dissolution. No pathological changes such as inflammatory cell infiltration, congestion, or edema were observed in the tissue. Compared with the normal group, the adrenal tissue morphology of the model group and the various drug-treated groups was similar: cells were densely arranged with clear layers, chromatin was evenly distributed, and no pathological changes such as inflammatory cell infiltration, congestion, or edema were observed in the tissue. These results indicate that the water-loaded polyuria model and the various drug-treated groups did not cause pathological damage to the adrenal tissue.

[0192] like Figure 6 As shown, the bladder tissue sections of animals in the normal group, water-loaded polyuria model group, and each drug-treated group showed intact mucosal layers with no shedding or fragmentation. The smooth muscle layer of the muscularis was tightly arranged and orderly, with no inflammatory infiltration and normal tissue morphology. This indicates that the water-loaded polyuria model and each drug-treated group did not cause pathological damage to bladder tissue.

[0193] In summary, the water-loaded polyuria model and the treatments in each drug-administered group did not cause pathological damage to the liver tissue, the urine metabolism and regulatory organs, and the kidney, adrenal gland, and bladder tissue.

[0194] Experimental studies have shown that the Suquan Pills (SQW) and high- and medium-dose Guben Yanling Pills groups prolonged urination latency compared to the water-loaded polyuria model group (M), delaying urination. Total urine output over 6 hours and its ratio to body weight were observed. Compared to the water-loaded polyuria model group (M), the Suquan Pills (SQW) and high-, medium-, and low-dose Guben Yanling Pills groups all reduced the total excretion rate and urine volume-to-weight ratio. The high-dose Guben Yanling Pills group showed a highly significant difference in urine volume-to-weight ratio. This suggests that both the positive drug Suquan Pills and the investigational drug Guben Yanling Pills have a certain degree of urinary reduction effect and should be able to improve the symptoms of patients with nocturia.

Claims

1. A Guben Yanling Pill for improving nocturnal urination, characterized in that: It is prepared from the following Chinese medicinal raw materials: ginseng, yam, cornus officinalis, akebia, wax gourd peel, dried tangerine peel, cypress seed, achyranthes bidentata, antler glue, tortoise shell glue, pyrrosia, licorice, Morinda officinalis, mulberry silkworm cocoon, Alpinia oxyphylla, Rosa laevigata, raspberry, and cat's whiskers.

2. The Guben Yanling Pill for improving nocturnal urination according to claim 1, characterized in that: The invention is prepared from the following Chinese medicinal raw materials, calculated by weight: 3-12 parts of ginseng, 5-15 parts of yam, 3-12 parts of cornus officinalis, 2-14 parts of akebia, 3-10 parts of wax gourd peel, 2-10 parts of dried tangerine peel, 1-8 parts of cypress seed, 3-12 parts of achyranthes bidentata, 1-7 parts of antler glue, 1-7 parts of tortoise shell glue, 5-16 parts of pyrrosia, 0.5-5 parts of liquorice, 3-10 parts of morinda officinalis, 4-15 parts of silkworm cocoon, 3-10 parts of alpinia oxyphylla, 6-20 parts of rose hip, 6-20 parts of raspberry, and 6-20 parts of cat's whiskers.

3. The Guben Yanling Pill for improving nocturnal urination according to claim 2, characterized in that: The invention is prepared from the following Chinese medicinal raw materials, calculated by weight: 4-8 parts of ginseng, 8-12 parts of yam, 4-10 parts of cornus officinalis, 4-10 parts of akebia, 4-8 parts of wax gourd peel, 4-8 parts of dried tangerine peel, 2-6 parts of cypress seed, 4-8 parts of achyranthes bidentata, 2-6 parts of antler glue, 2-6 parts of tortoise shell glue, 6-12 parts of pyrrosia, 1-4 parts of liquorice, 4-8 parts of morinda officinalis, 6-12 parts of silkworm cocoon, 4-8 parts of alpinia oxyphylla, 8-16 parts of rose hip, 8-16 parts of raspberry, and 8-16 parts of cat's whiskers.

4. The Guben Yanling Pill for improving nocturnal urination according to claim 3, characterized in that: The invention is prepared from the following Chinese medicinal raw materials, calculated by weight: 6 parts of ginseng, 10 parts of yam, 8 parts of cornus officinalis, 8 parts of akebia, 6 parts of wax gourd peel, 6 parts of dried tangerine peel, 4 parts of cypress seed, 6 parts of achyranthes bidentata, 4 parts of antler glue, 4 parts of tortoise shell glue, 9 parts of pyrrosia, 3 parts of liquorice, 6 parts of morinda officinalis, 9 parts of silkworm cocoon, 6 parts of alpinia oxyphylla, 10 parts of rose hip, 10 parts of raspberry, and 10 parts of cat's whiskers.

5. The method for preparing the Guben Yanling Pill for improving nocturnal urination according to any one of claims 1 to 4, characterized in that: The steps include: (1) Weigh the above raw materials in order by weight; (2) The Chinese medicinal materials except deer antler glue and tortoise shell glue are crushed and mixed, cellulase and water are added for enzymatic extraction, followed by ultrasonic-assisted extraction, and the first filtrate is obtained by filtration; Adding ethanol aqueous solution to the precipitate for secondary extraction, filtering to obtain a second filtrate; combining the two filtrates and concentrating to obtain a Chinese medicine paste; (3) extracting and concentrating the deer antler glue and tortoise shell glue with water to obtain a thick paste; (4) Mix the Chinese medicine paste and thick paste, add a binder to make Guben Yanling Pills to improve frequent urination at night.

6. The method for preparing Guben Yanling Pills for improving nocturnal urination according to claim 5, characterized in that: The amount of cellulase added is 0.5-2wt% of the total mass of the raw materials, the enzymatic hydrolysis pH is 3-5, the temperature is 40-60°C, the stirring speed is 200-600rpm, and the time is 30-60min.

7. The method for preparing Guben Yanling Pills for improving nocturnal urination according to claim 5, characterized in that: The ultrasonic extraction conditions are: power 50-150W, frequency 20-60kHz, temperature 60-90°C, and time 1-3h.

8. The method for preparing Guben Yanling Pills for improving nocturnal urination according to claim 5, characterized in that: The concentration of the ethanol aqueous solution is 40-60 wt %, the extraction temperature is 70-85° C., and the extraction time is 2-4 h.

9. The method for preparing Guben Yanling Pills for improving nocturnal urination according to claim 5, characterized in that: The extraction conditions in step (3) are: extraction at 70-90° C. for 1-3 times, with each extraction time being 0.5-2 h.

10. The method for preparing Guben Yanling Pills for improving nocturnal urination according to claim 5, characterized in that: The binder is one of honey, starch or hydroxypropyl methylcellulose, and the addition amount is 2-6wt% of the total paste mass.

Images