A medicinal and edible Chinese medicine composition for treating insomnia and a preparation method thereof

Abstract

This invention provides a traditional Chinese medicine formula for treating insomnia that can be used as both food and medicine, and its preparation method, belonging to the field of traditional Chinese medicine preparation technology. The traditional Chinese medicine formula of this invention includes: 2-8 parts egg yolk, 1-7 parts donkey-hide gelatin, 2-8 parts jujube seed, 1-7 parts Solomon's seal rhizome, 2-8 parts lily bulb, 1-7 parts longan pulp, 2-8 parts ophiopogon root, 2-8 parts angelica root, 1-7 parts clove, 3-10 parts dried tangerine peel, 1-7 parts mulberry leaf, 1-7 parts chrysanthemum, 2-8 parts peach kernel, 4-13 parts kudzu root, 1-7 parts saffron, 2-8 parts ganoderma lucidum, 2-8 parts cistanche, 2-8 parts raspberry, 2-8 parts wolfberry, 2-8 parts yam, 4-13 parts roasted malt, 1-7 parts roasted ginkgo kernel, 4-13 parts roasted euryale seed, and 2-8 parts prepared licorice root. The traditional Chinese medicine formula provided by this invention has certain effects on improving and treating insomnia, menopausal symptoms in women, and ovarian function decline. It is safe, effective, has no side effects, and has a long-lasting effect.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine composition technology, and in particular to a traditional Chinese medicine formula for treating insomnia that can be used as both food and medicine, and its preparation method. Background Technology

[0002] Sleep is essential for normal life and study; good sleep helps eliminate fatigue and enhance immunity. Insomnia is a common clinical condition characterized by dissatisfaction with sleep duration or quality, accompanied by daytime social dysfunction. Insomnia can induce or aggravate neuroinflammation, damage the hippocampus, and affect normal learning and memory functions. Long-term insomnia also increases the risk of cognitive impairment. Therefore, inhibiting neuroinflammation and protecting nerves is of great significance in improving anxiety caused by insomnia.

[0003] Most traditional Chinese medicines exhibit synergistic effects with sodium pentobarbital when exerting sedative and hypnotic effects. By reducing the number of spontaneous bodily activities, prolonging the suprathreshold and subthreshold sodium pentobarbital sleep time, and shortening the sleep latency time induced by sodium pentobarbital, they inhibit the function of the central nervous system, thereby suppressing bodily arousal, prolonging bodily sleep time, and improving bodily sleep quality.

[0004] Studies have shown that sleep and inflammatory responses are closely related. Cytokines can regulate the sleep process in the central nervous system through different pathways. Inflammatory cytokines such as interleukin (IL) and tumor necrosis factor (TNF) are important factors involved in sleep regulation. TNF-α, as an inflammatory cytokine, can promote the production and secretion of IL-1β, IL-2, and IL-6, inducing inflammatory responses. IL-10 is an anti-inflammatory factor that downregulates inflammatory responses and antagonizes inflammatory mediators. IL-1β, as an inflammatory factor with broad biological activity, can indirectly reflect sleep status. IL-2 and IL-6, as cytokines, participate in the regulation of sleep and immune function in the human body. Chronic insomnia often causes chronic inflammatory responses and changes in immune function, and altering the secretion levels of IL-2 and IL-6 plays an important regulatory role in insomnia. Research has found that the diurnal variations of other cytokines such as TNF-α, IL-2, IL-6, and IL-1β are closely related to sleep. Reduced sleep can increase the secretion of IL-2, IL-6, IL-1β and TNF-α, and decrease the secretion of the anti-inflammatory factor IL-10. TNF-α also plays an important role in autonomous life activities such as sleep and eating. Summary of the Invention

[0005] In view of this, the present invention provides a traditional Chinese medicine formula for treating insomnia that can be used as both food and medicine. The traditional Chinese medicine formula is composed of the following raw materials in parts by weight: 2-8 parts egg yolk, 1-7 parts donkey-hide gelatin, 2-8 parts jujube seed, 1-7 parts Solomon's seal rhizome, 2-8 parts lily bulb, 1-7 parts longan pulp, 2-8 parts ophiopogon root, 2-8 parts angelica root, 1-7 parts clove, 3-10 parts dried tangerine peel, 1-7 parts mulberry leaf, 1-7 parts chrysanthemum, 2-8 parts peach kernel, 4-13 parts kudzu root, 1-7 parts saffron, 2-8 parts ganoderma lucidum, 2-8 parts cistanche deserticola, 2-8 parts raspberry, 2-8 parts wolfberry, 2-8 parts yam, 4-13 parts roasted malt, 1-7 parts roasted ginkgo kernel, 4-13 parts roasted euryale seed, and 2-8 parts roasted licorice root.

[0006] Furthermore, the traditional Chinese medicine formula consists of the following ingredients in parts by weight: 2 parts egg yolk, 1 part donkey-hide gelatin, 2 parts jujube seed, 1 part Solomon's seal rhizome, 2 parts lily bulb, 1 part longan pulp, 2 parts ophiopogon root, 2 parts angelica root, 1 part clove, 3 parts dried tangerine peel, 1 part mulberry leaf, 1 part chrysanthemum, 2 parts peach kernel, 4 parts kudzu root, 1 part saffron, 2 parts ganoderma lucidum, 2 parts cistanche, 2 parts raspberry, 2 parts wolfberry, 2 parts yam, 4 parts roasted malt, 1 part roasted ginkgo kernel, 4 parts roasted foxnut, and 2 parts roasted licorice root.

[0007] Furthermore, the dosage forms of the traditional Chinese medicine formulation include ointments, granules, and tablets.

[0008] Furthermore, the preparation method of the ointment includes the following steps: weigh each raw material according to the proportion, wash and dry them, then pulverize them to 100-150 mesh, then mix the obtained powder, add 8-10 times the total weight of pure water of the raw materials, extract for 1-3 hours, filter to obtain extract a and residue a; add 8-10 times the amount of pure water to residue a, extract for 1-3 hours, filter to obtain extract b and residue b; mix extract a and extract b, add 40-60% honey, and concentrate to 1.3 times the concentration to obtain the ointment.

[0009] Furthermore, the preparation method of the granules includes the following steps: weigh each raw material according to the proportion, wash and dry them, then pulverize them to 100-150 mesh, then mix the obtained powder, add 8-10 times the total weight of pure water to the raw materials, extract for 1-3 hours, filter to obtain extract a and residue a; add 8-10 times the amount of pure water to residue a, extract for 1-3 hours, filter to obtain extract b and residue b; mix extract a and extract b, concentrate, spray dry to obtain extract powder, then add the extract powder to edible ethanol, mix, dry and granulate in sequence to obtain granules.

[0010] Furthermore, the preparation method of the tablets includes the following steps: weighing each raw material according to the proportion, washing and drying them, then pulverizing them to 100-150 mesh, mixing the obtained powders, adding 8-10 times the total weight of pure water to the raw materials, extracting for 1-3 hours, filtering to obtain extract a and residue a; adding 8-10 times the amount of pure water to residue a, extracting for 1-3 hours, filtering to obtain extract b and residue b; mixing extract a and extract b, concentrating, spray drying to obtain extract powder, adding the extract powder to edible ethanol, mixing, drying, and granulating in sequence to obtain granules, adding excipients to the granules, compressing into tablets to obtain tablets.

[0011] Furthermore, the excipients include xylooligosaccharides, magnesium stearate, calcium sulfate, sugar powder, lactose, or any combination thereof, and the excipients constitute 0.1-3% of the total weight of the granules.

[0012] Furthermore, the extraction temperature is 60-90℃.

[0013] The ingredients used in the medicinal and edible herbal formula for treating insomnia described in this invention are explained below:

[0014] 1. Egg yolk can nourish the heart and calm the mind, and replenish blood and yin; donkey-hide gelatin can replenish blood and stop bleeding, and nourish yin and moisten dryness. The combination of the two has a good effect on symptoms such as blood deficiency insomnia and palpitations.

[0015] 2. Sour jujube seed is a medicine for nourishing the heart and calming the mind, mainly used to treat insomnia, palpitations, and excessive dreaming due to deficiency and restlessness; Polygonatum odoratum has the effects of nourishing yin, moistening dryness, clearing heat, and promoting body fluid production, and can improve cardiovascular function. Lily bulb can nourish yin and moisten the lungs, clear the heart and calm the mind. Modern pharmacological studies have shown that it has a significant sedative effect and can withstand hypoxia and fatigue. The combination of these three has a synergistic effect on symptoms such as restlessness, insomnia, and excessive dreaming.

[0016] 3. Longan pulp has the effects of nourishing blood and qi, eliminating fatigue, and calming the mind; Ophiopogon japonicus contains rich protein, vitamins, and amino acids, and has the effects of nourishing yin and promoting body fluids, delaying aging, moistening the lungs and clearing the heart, moistening the intestines and promoting bowel movements, and aiding sleep. The combination of the two is suitable for improving hot flashes and night sweats, qi and blood deficiency, neurasthenia, and poor sleep quality caused by menopause.

[0017] 4. Angelica dahurica can dispel wind and relieve pain, clear the orifices and release the exterior. It is often used to treat headaches, nasal congestion and other symptoms. Clove has the effects of moistening the intestines and relieving constipation, warming the kidneys and strengthening yang, and relieving insomnia. Tangerine peel can regulate qi and strengthen the spleen, dry dampness and resolve phlegm. It has a good effect on spleen and stomach disharmony and phlegm and dampness obstruction. The hesperidin it contains is a natural antioxidant and has a certain anti-arteriosclerosis effect.

[0018] Mulberry leaves are primarily used to dispel wind and clear heat, clear the liver and improve eyesight, and relieve symptoms such as irritability, anger, and excessive liver fire.

[0019] Chrysanthemum has the effects of clearing heat and detoxifying, and calming liver yang.

[0020] Peach kernels promote blood circulation, remove blood stasis, moisten the intestines, and relieve constipation;

[0021] Kudzu root can relieve muscle tension and reduce fever, promote body fluid production and quench thirst, and raise yang energy to stop diarrhea;

[0022] Saffron promotes blood circulation, removes blood stasis, relieves depression, and calms the mind. When the above eight ingredients are used together, they can regulate qi and strengthen the spleen, moisten the intestines and promote bowel movements, relieve irritability, relieve muscle tension and reduce fever, and relieve depression and calm the mind.

[0023] 5. Ganoderma lucidum can replenish qi and calm the mind, relieve cough and asthma, and is often used for symptoms such as restlessness and insomnia;

[0024] Cistanche deserticola tonifies kidney yang, nourishes essence and blood, and moistens the intestines to relieve constipation. It has a good effect on kidney yang deficiency and essence and blood deficiency.

[0025] Raspberries contain natural estrogen and have the effect of nourishing the kidneys and replenishing essence, as well as regulating female estrogen levels.

[0026] Goji berries have benefits including nourishing the liver and kidneys, and clearing the liver and improving eyesight.

[0027] Yam has multiple benefits, including strengthening the spleen, nourishing the lungs, tonifying the kidneys, and replenishing essence. Yam is rich in estrogen, which helps improve ovarian function and increase estrogen levels. The combination of these five ingredients can replenish qi and calm the mind, tonify the kidneys and replenish essence, improve ovarian function, and increase estrogen levels.

[0028] 6. Roasted malt can promote digestion, strengthen the spleen and stimulate appetite; roasted ginkgo kernels can relieve cough and asthma, and astringe and stop leukorrhea; roasted fox nuts can benefit the kidneys and strengthen essence, and tonify the spleen and stop diarrhea. When used together, these three ingredients can strengthen the spleen and stimulate appetite, as well as astringe and stop leukorrhea. They are effective for symptoms such as spleen and stomach weakness and excessive leukorrhea.

[0029] 7. Prepared licorice root can invigorate the spleen and stomach, replenish qi and restore pulse, relieve spasms and pain, and harmonize the effects of other medicines.

[0030] The combined effects of these herbs are to nourish blood and calm the mind, nourish yin and moisten dryness, strengthen the spleen and stomach, promote blood circulation and remove blood stasis, and tonify the kidneys and consolidate essence.

[0031] This formula is mainly used to treat symptoms such as insomnia, excessive dreaming, palpitations, dizziness, abdominal distension, and numbness of limbs caused by disharmony between the heart and kidneys, disharmony between the spleen and stomach, blood stasis and phlegm obstruction.

[0032] The medicinal and edible herbal formula provided by this invention has certain improving and therapeutic effects on insomnia, menopausal symptoms in women, and ovarian function decline. It is safe, effective, has no side effects, and provides long-lasting results. This herbal formula does not limit the dosage form or product type; it can be made into compressed candies, pastes, granules, meal replacement powders, and other health products. These are convenient to consume, easy to promote, and widely applicable in medical institutions, health management centers, and daily family dietary supplements, possessing significant social and healthcare value. Attached Figure Description

[0033] Figure 1 This is a graph showing the effect of the ointment of the present invention on the body weight of PCPA-induced insomnia mice.

[0034] Figure 2 These are open field behavioral trajectory diagrams of mice in each group according to the present invention.

[0035] Figure 3 These are behavioral trajectory diagrams of the elevated cross maze for each group of mice in this invention.

[0036] Figure 4 This is a diagram showing the results of the observation of the neuropathological morphology of the CA1, DG, and CA3 regions of the mouse hippocampus using the ointment of this invention.

[0037] Figure 5 These are images of the neuropathological morphology (hematoxylin-eosin staining, ×400) of the CA1 region of the hippocampus in mice of each group in this invention.

[0038] Figure 6 These are images of the neuropathological morphology (hematoxylin-eosin staining, ×400) of the hippocampal DG region in mice of each group according to the present invention.

[0039] Figure 7 These are images showing the neuropathological morphology (hematoxylin-eosin staining, ×400) of the CA3 region of the hippocampus in mice from each group of mice in this invention.

[0040] Figure 8 This is a graph showing the effect of the ointment of the present invention on the levels of IL-2, IL-6 and IL-10 in mouse serum.

[0041] Figure 9 This is a graph showing the effect of the ointment of the present invention on the levels of TNF-α and IL-1β in mouse serum. Detailed Implementation

[0042] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of this invention, and not all of them. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0043] Unless otherwise specified, the test methods or experimental methods described in the following examples are all conventional methods; unless otherwise specified, the raw materials and additives are obtained from conventional commercial sources or prepared by conventional methods.

[0044] Example 1

[0045] A medicinal and edible herbal ointment for treating insomnia.

[0046] Prescription: 2 parts egg yolk, 1 part donkey-hide gelatin, 2 parts jujube seed, 1 part Solomon's seal rhizome, 2 parts lily bulb, 1 part longan pulp, 2 parts ophiopogon root, 2 parts angelica root, 1 part clove, 3 parts dried tangerine peel, 1 part mulberry leaf, 1 part chrysanthemum, 2 parts ganoderma lucidum, 2 parts peach kernel, 4 parts kudzu root, 1 part saffron, 2 parts cistanche, 2 parts raspberry, 2 parts wolfberry, 2 parts yam, 4 parts roasted malt, 1 part roasted ginkgo kernel, 4 parts roasted foxnut, 2 parts prepared licorice root.

[0047] Preparation process:

[0048] Weigh each raw material according to the proportion, wash and dry them, then pulverize them to 100 mesh. After sieving, mix the resulting powders, add 8 times the total weight of pure water, extract at 60℃ for 1 hour, filter, and obtain extract a and residue a; add 8 times the amount of pure water to residue a, extract at 60℃ for 1 hour, filter, and obtain extract b and residue b; mix extract a and extract b, add 50% honey, and concentrate to 1.3 times the concentration (50℃) to obtain an ointment.

[0049] Example 2

[0050] A granule formulation of traditional Chinese medicine that can be used as both food and medicine to treat insomnia.

[0051] Prescription: 8 parts egg yolk, 7 parts donkey-hide gelatin, 8 parts jujube seed, 7 parts Solomon's seal rhizome, 8 parts lily bulb, 7 parts longan pulp, 8 parts ophiopogon root, 8 parts angelica root, 7 parts clove, 10 parts dried tangerine peel, 7 parts mulberry leaf, 7 parts chrysanthemum, 8 parts peach kernel, 13 parts kudzu root, 7 parts saffron, 8 parts ganoderma, 8 parts cistanche, 8 parts raspberry, 8 parts wolfberry, 8 parts yam, 13 parts roasted malt, 7 parts roasted ginkgo kernel, 13 parts roasted foxnut, 8 parts prepared licorice root.

[0052] Preparation process: Weigh each raw material according to the proportion, wash and dry them, then pulverize them to 120 mesh, sieve them, mix the resulting powders, add 9 times the total weight of pure water, extract at 80℃ for 2 hours, filter to obtain extract a and residue a; add 9 times the amount of pure water to residue a, extract at 80℃ for 2 hours, filter to obtain extract b and residue b; mix extract a and extract b, concentrate to 1.3 times the concentration (50℃), spray dry to obtain extract powder, then add the extract powder to an appropriate amount of edible ethanol, mix, dry and granulate in sequence to obtain granules.

[0053] Example 3

[0054] A tablet made from a traditional Chinese medicine formula that can be used as both food and medicine to treat insomnia.

[0055] Prescription: 6 parts egg yolk, 5 parts donkey-hide gelatin, 6 parts jujube seed, 5 parts Solomon's seal rhizome, 6 parts lily bulb, 5 parts longan pulp, 6 parts ophiopogon root, 6 parts angelica root, 5 parts clove, 8 parts dried tangerine peel, 5 parts mulberry leaf, 5 parts chrysanthemum, 7 parts peach kernel, 8 parts kudzu root, 5 parts saffron, 6 parts ganoderma lucidum, 7 parts cistanche, 7 parts raspberry, 7 parts wolfberry, 6 parts yam, 8 parts roasted malt, 5 parts roasted ginkgo kernel, 8 parts roasted foxnut, 6 parts prepared licorice root.

[0056] Preparation process: Weigh each raw material according to the proportion, wash and dry them, then pulverize them to 150 mesh, sieve them, mix the resulting powders, add 10 times the total weight of pure water, extract at 90℃ for 3 hours, filter to obtain extract a and residue a; add 10 times the amount of pure water to residue a, extract at 90℃ for 3 hours, filter to obtain extract b and residue b; mix extract a and extract b, concentrate to 1.3 times the concentration (50℃), spray dry to obtain extract powder, add the extract powder to an appropriate amount of edible ethanol, mix, dry and granulate in sequence to obtain granules, add 1% excipients to the granules, compress into tablets to obtain tablets.

[0057] I. Animal Pharmacological Experiments

[0058] 1. Grouping and treatment of laboratory animals

[0059] One hundred healthy male ICR mice were acclimatized for 5 days and then randomly divided into two batches of 50 mice each. The first batch underwent a pentobarbital sodium-induced sleep experiment, while the second batch underwent behavioral experiments and serum marker measurements. Each batch of animals was further divided into a control group, a model group (PCPA, 350 mg / kg), a positive control group (diazepam, 2.6 mg / kg), and low- and high-dose groups of the ointment from Example 1 (0.9 g / kg and 1.8 g / kg, respectively). The control group and the PCPA group received an equal volume of physiological saline, while the other groups received the corresponding drugs. The sleep experiment was conducted via gavage for 37 consecutive days, and the open field experiment for 38 consecutive days. Body weight was measured weekly.

[0060] 2. Establishment of a mouse model of insomnia induced by p-chlorophenylalanine (PCPA)

[0061] From day 29 to day 31 after drug administration, the control group received intraperitoneal injections of physiological saline, while the PCPA group, diazepam group, and the low- and high-dose ointment groups of Example 1 received intraperitoneal injections of PCPA 350 mg / kg once daily for 3 consecutive days. When the model mice exhibited loss of circadian rhythm, increased irritability and aggression, and showed significant differences from the control group, the model was successfully replicated. This model generally lasts for about 2 weeks. The preparation process of the PCPA suspension was as follows: accurately weigh 1.0 g of PCPA, dissolve it in 30 mL of physiological saline, add 20 μL of Tween 80 to promote dissolution, and vortex to obtain the PCPA suspension.

[0062] 3. Sodium pentobarbital-induced sleep experiment in mice

[0063] 3.1 Direct Sleep Experiment

[0064] Each group of animals was given the corresponding drug daily, and the mice were observed to exhibit sleep patterns within 60 minutes after gavage. Sleep was defined as the absence of the righting reflex for more than 60 seconds, and awakening was defined as the return of the righting reflex. The number of animals falling asleep and the duration of sleep were recorded for each group to observe whether the test substance had a direct sleep effect on the mice.

[0065] 3.2 Subthreshold dose hypnotic experiment with sodium pentobarbital

[0066] Thirty minutes after the last gavage, sodium pentobarbital was injected intraperitoneally at a dose of 30 mg / kg (based on mouse body weight, the dose for 80%–90% of mice that did not fall asleep). The number of animals that fell asleep within 30 minutes of the start of intraperitoneal injection was recorded, with the disappearance of the righting reflex for 60 seconds as the indicator.

[0067] 3.3 Pentobarbital sodium sleep latency and sleep duration experiment

[0068] Twenty minutes after the last gavage, sodium pentobarbital was injected intraperitoneally at a dose of 45 mg / kg (based on mouse body weight, ensuring 100% sleep in mice). The sleep latency was defined as the time between intraperitoneal injection and the disappearance of the righting reflex, and the sleep duration was defined as the time between the disappearance and recovery of the righting reflex. The differences in sleep latency and sleep duration between the treatment group and the model control group were compared.

[0069] 4. Behavioral tests

[0070] 4.1 Open Field Experiment

[0071] On day 38 after drug administration, an open field test was conducted. The open field test is a method for evaluating the autonomous behavior, exploratory behavior, and stress levels of mice in a new environment. Sixty minutes after the last drug administration, all experimental animals underwent behavioral testing using the open field test. The open field test chamber was 50 cm high and 50 cm long at the base, with a video acquisition device at the top. Mice were placed in the center of the chamber, and their activity was recorded for 5 minutes using a mouse movement trajectory behavior analysis system. The experiment was conducted in a quiet environment with stable lighting. After each mouse's experiment, the inner walls and bottom of the chamber were wiped with 75% alcohol to prevent the retention of odors and excrement. Test indicators included: total distance traveled in the open field, total resting time, central dwell time, number of standing movements, number of grooming movements, and number of fecal particles.

[0072] 4.2 Elevated Cross Maze Experiment

[0073] After the open field experiment, mice in each group underwent the elevated cross maze experiment. The entire experiment was recorded using video recording equipment configured for the elevated cross maze, and behavioral software was used to record the total number of arm insertions, total arm insertion time, number of open arms (OE), open arm dwell time (OT), number of closed arms (CE), and closed arm time (CT) for each group of mice within 5 minutes. The environment was kept quiet during the experiment, and mouse excrement was cleaned up promptly after each test and wiped with 75% alcohol.

[0074] 4.3 Sugar Water Preference Experiment

[0075] After modeling and gavage, sucrose water acclimatization training was conducted, with water deprivation for 24 hours. Then, one bottle of 1% sucrose solution and one bottle of pure water were placed in each cage (first check if the water bottles were dripping). After 2 hours, both bottles were removed and weighed. The consumption of sucrose water and pure water was recorded, and the sucrose water preference rate was calculated.

[0076] Sucrose water preference rate = sucrose water consumption / (sucrose water consumption + pure water consumption) × 100%.

[0077] 5. Serum marker testing

[0078] One hour after the last administration, mice in each group were anesthetized with sodium pentobarbital (30 mg / kg) via intraperitoneal injection. Blood was collected from the abdominal aorta, centrifuged for 15 minutes (4000 rpm), and serum was separated. Mouse serum was collected and stored at -80℃ for later use. The levels of interleukin-2 (IL-2), interleukin-6 (IL-6), interleukin-10 (IL-10), interleukin-1β (IL-1β), and α-tumor necrosis factor (TNF-α) in the serum of each group of mice were determined using enzyme-linked immunosorbent assay (ELISA). The assays were performed strictly according to the instructions provided with the ELISA kit.

[0079] 6. Morphological observation of hippocampal tissue

[0080] HE staining was used to detect pathological changes in mouse hippocampal tissue. Hippocampal tissue samples from each group of mice were fixed in 4% paraformaldehyde for 2 hours. After rinsing, the fixed tissue samples were dehydrated and then routinely embedded in paraffin to prepare paraffin sections of about 4 μm. After dewaxing, the sections were washed stepwise with graded alcohols until they became clear. After staining with hematoxylin-eosin, the sections were dehydrated, mounted with neutral resin, and the morphological changes of hippocampal cells were observed under an optical microscope. Target areas were photographed and analyzed.

[0081] 7 Data Processing

[0082] SPSS 24 and Origin 2021 software were used for statistical analysis and plotting of the data. Tukey's test was used for pairwise comparisons between multiple group means. Experimental data are expressed as mean ± standard deviation. The expression indicates that a p-value < 0.05 is considered statistically significant.

[0083] 8. Test Results

[0084] 8.1 The effect of the ointment in Example 1 of this invention on the body weight of PCPA-induced insomnia mice is shown in [the figure]. Figure 1 .

[0085] Depend on Figure 1 It was found that the control group mice were agile, in good spirits, and had normal activity, sleep, diet, and body weight. Their fur was glossy and smooth, and they exhibited a clear sleep rhythm, showing nocturnal activity. Compared to the control group, the PCPA group mice showed poor spirits, reduced activity and diet, duller fur, and slower weight gain within one week of modeling. Compared to the PCPA group, the diazepam group and the different dosage groups in Example 1 showed less mutual aggression and biting within one week of administration, had cleaner fur, normal diet, and continued weight gain.

[0086] 8.2 The effect of the ointment in Example 1 of the present invention on direct sleep in mice is shown in Table 1. (n=10).

[0087] Table 1 shows that this experiment observed the activity of mice in each group within 60 minutes after gavage. All mice exhibited normal activity, with no loss of righting position or sleep patterns observed; the sleep onset rate and sleep duration were both 0. This indicates that the sleep quality of the mice in Example 1 was improved during normal sleep, rather than by inducing a hypnotic state. This suggests that further evaluation can be conducted in subsequent experiments.

[0088] Table 1

[0089]

[0090]

[0091] 8.3 The effects of the ointment of Example 1 of the present invention on sleep time and sleep latency in PCPA-induced insomnia mice are shown in Table 2. (n=10).

[0092] As shown in Table 2, compared with the blank group, the sleep rate of mice in the PCPA group was reduced (P<0.05). Compared with the PCPA group, the sleep rate of mice in each dose group of Example 1 was significantly increased (P<0.05, P<0.001), indicating that the traditional Chinese medicine formula of the present invention can improve the sleep rate of mice, and the sleep rate is dose-dependent.

[0093] Table 2

[0094]

[0095] Note: Compared with the blank group, "###" indicates P < 0.001, "##" indicates P < 0.01, and "#" indicates P < 0.05; compared with the model group, "***" indicates P < 0.001, "**" indicates P < 0.01, and "*" indicates P < 0.05. The same applies below.

[0096] 8.4 The effects of the ointment of Example 1 of the present invention on the open field behavior of PCPA-induced insomnia mice are shown in Table 3. (n=10).

[0097] Table 3 shows that compared with the control group, the PCPA group mice exhibited a highly prolonged sleep latency induced by sodium pentobarbital (P < 0.05) and a significantly reduced sleep duration (P < 0.05), indicating increased excitability and successful model establishment. Compared with the PCPA group, the diazepam group mice showed a significantly prolonged sleep duration (P < 0.001) and a highly prolonged sleep latency (P < 0.001). The high-dose group of Example 1 mice showed a significantly shortened sleep latency (P < 0.01), indicating a dose-response relationship between the groups and suggesting that Example 1 has an improving effect on sleep in insomniac mice.

[0098] Table 3

[0099]

[0100]

[0101] The open field behavioral trajectories of mice in each group are shown below. Figure 2 .

[0102] 8.5 The effects of the ointment of Example 1 of the present invention on the behavior of PCPA-induced insomnia mice in the elevated cruciate maze are shown in Table 4. (n=10).

[0103] From Table 4 and Figure 2 It was found that compared with the control group, the total movement distance and center dwell time of mice in the PCPA group were significantly reduced (P<0.01, P<0.05), and the total resting time was significantly increased (P<0.05). The number of standing, modification, and fecal pellet counts were also significantly increased (P<0.05, P<0.01, P<0.01). Compared with the PCPA group, the total movement distance of mice in the diazepam group was significantly increased (P<0.01), and the total resting time and fecal pellet count were significantly reduced (P<0.01). The total resting time of mice in the high-dose group of Example 1 was significantly reduced (P<0.01). The total movement distance and center dwell time of mice in the high- and low-dose groups showed an increasing trend compared with the PCPA group, but this was not statistically significant. The number of modification counts of mice in the low-dose group was significantly reduced (P<0.05), and the number of standing, modification, and fecal pellet counts of mice in the high-dose group were all significantly reduced (P<0.05, P<0.001, P<0.05).

[0104] Table 4

[0105]

[0106] The behavioral trajectories of the mice in the elevated cross maze for each group are shown below. Figure 3 .

[0107] 8.6 The effect of the ointment of Example 1 of the present invention on the sugar water preference rate of PCPA-induced insomnia mice is shown in Table 5. (n=10).

[0108] From Table 5 and Figure 3 It was found that compared with the control group, the PCPA group mice had significantly reduced percentages of open-arm retention time, open-arm movement distance, number of times entering the open arm, and number of times the open arm probe was used (P < 0.05); compared with the PCPA group, the diazepam group mice had significantly increased percentages of open-arm retention time, open-arm movement distance, number of times entering the open arm, and number of times the open arm probe was used (P < 0.05, P < 0.01, P < 0.05, P < 0.05), and the differences were statistically significant. In Example 1, the low-dose and high-dose groups mice had significantly increased percentages of open-arm retention time and number of times entering the open arm (P < 0.05), and the high-dose group mice had significantly increased percentages of open-arm movement distance and number of times the open arm probe was used (P < 0.05, P < 0.01).

[0109] Table 5

[0110]

[0111] 8.7 The results of the observation of the neuropathological morphology of the CA1, DG, and CA3 regions of the mouse hippocampus by the ointment of Example 1 of the present invention are shown in the figure. Figure 4 Neuropathological morphology of the CA1, DG, and CA3 regions of the hippocampus in each group of mice (hematoxylin-eosin staining, ×400) is shown in the figure. Figure 5 , Figure 6 and Figure 7 .

[0112] We assessed the histopathological changes in the morphology and structure of the CA1, DG, and CA3 regions of the mouse hippocampus after PCPA induction using HE staining. Figure 4-5HE staining results showed that, compared with the control group, the number of neurons in the CA1, DG, and CA3 regions of the hippocampus of mice in the PCPA group was significantly reduced (P < 0.05), with increased intercellular spaces, nuclear pyknosis, and an increased number of degenerated and necrotic cells. Compared with the PCPA group, the number of neurons in all regions of the hippocampus of mice in each dose group of Example 1 showed an increasing trend. In the DG region, the number of neurons in both the low- and high-dose groups of Example 1 was significantly increased (P < 0.05, P < 0.01); in the CA3 region, the number of neurons in the high-dose group of Example 1 was significantly increased (P < 0.05). The cells in the drug-treated groups were neatly arranged and clearly visible, with increased layers and reduced nuclear pyknosis.

[0113] Figure 6 Compared with the control group, the serum IL-2 and IL-6 levels in the PCPA group were significantly increased (P < 0.05), and the IL-10 level was significantly decreased (P < 0.05). Compared with the PCPA group, the serum IL-2 and IL-6 levels in the diazepam group were significantly decreased (P < 0.01, P < 0.05), and the serum IL-2 and IL-6 levels in the high-dose group were significantly decreased (P < 0.05). Although there was no statistically significant difference in serum IL-10 levels among the groups in Example 1 compared with the PCPA group (P > 0.05), all showed an increasing trend.

[0114] Figure 7 Compared with the control group, the serum TNF-α and IL-1β levels in the PCPA group mice were significantly increased, and the differences were statistically significant (P < 0.01, P < 0.05). Compared with the PCPA group, the serum TNF-α levels in the diazepam group and the high-dose group mice were significantly decreased (P < 0.01), and the serum IL-1β levels in the low-dose group and the high-dose group mice were significantly decreased (P < 0.01).

[0115] Table 6: Compared with the control group, the sucrose preference rate of mice in the PCPA group was significantly reduced (P<0.05); compared with the PCPA group, the sucrose consumption and sucrose preference rate of mice in the diazepam group were significantly increased (P<0.001), the sucrose consumption and sucrose preference rate of mice in the low-dose group of Example 1 were significantly increased (P<0.05, P<0.01), and the sucrose consumption and sucrose preference rate of mice in the high-dose group were significantly increased (P<0.001). There was a dose-dependent relationship among the dose groups.

[0116] 8.8 The effect of the ointment in Example 1 of this invention on the levels of IL-2, IL-6 and IL-10 in mouse serum is shown in [reference needed]. Figure 8 .

[0117] 8.9 The effect of the ointment in Example 1 of this invention on the levels of TNF-α and IL-1β in mouse serum is shown in [reference needed]. Figure 9 .

[0118] 9. Conclusion

[0119] Mice deprived of sleep exhibited disrupted circadian rhythms, displaying negative emotions such as anxiety and aggression, and showed slow weight gain. Compared to the control group, the PCPA group showed a significant increase in wakefulness time. Different doses of the ointment from Example 1 alleviated the mental state of sleep-deprived mice, resulting in rapid weight gain, reduced wakefulness time, and a significant increase in total sleep duration. The sodium pentobarbital test results showed that the ointment from Example 1 significantly increased the number of sleep episodes in mice at subthreshold doses of sodium pentobarbital and shortened the sleep latency induced by sodium pentobarbital. All results were positive. Based on the sleep-improving function testing methods in the "Technical Specifications for Inspection and Evaluation of Health Foods," the ointment from Example 1 can be determined to have a sleep-improving function.

[0120] Meanwhile, this invention used open field test, elevated cross maze test, and sucrose preference test to study the emotional changes and anxiety levels of mice. In the PCPA group, the total activity distance and central dwell time in the open field were significantly reduced, while the rest time increased, indicating that sleep deprivation disrupted the diurnal rhythm, caused anxiety, and reduced cognitive ability in new environments. The ointment group in Example 1 improved their anxiety state, with the high-dose group showing the most significant difference and activity levels approaching those of the control group. In the elevated cross maze test, the percentage of time spent in the open arm and the percentage of times entering the open arm were increased in the Ointment group of Example 1. In the sucrose preference test, the sucrose preference rate of the PCPA group was significantly lower than that of the control group, indicating that sleep deprivation led to a loss of sucrose preference due to insufficient pleasure, a core symptom of depression. The high-dose Ointment group of Example 1 showed a significantly increased sucrose preference rate, indicating that depressive mood was fully alleviated and pleasure was restored.

[0121] The results of this study showed that, compared with the control group, the PCPA group mice had significantly higher serum expression levels of IL-2, IL-6, IL-1β, and TNF-α. The hippocampal neurons in these mice exhibited irregular arrangement, widened intercellular spaces, nuclear pyknosis, and an increased number of degenerated and necrotic cells. This suggests that the ointment in Example 1 may improve insomnia symptoms by regulating inflammatory factor levels and protecting brain tissue neurons.

[0122] In summary, the ointment in Example 1 can effectively treat insomnia and improve anxiety caused by insomnia. The mechanism mediated by this ointment may be related to inhibiting neuroinflammation and reducing nerve damage.

[0123] II. Clinical Trials

[0124] 1. Case selection

[0125] Volunteers with eligible menopausal syndrome who visited Hebei Port Hospital between January and December 2023 were randomly divided into two groups: one group received oral administration of the tablets described in Example 3 of this invention, and the other group received oral administration of the tablets described in Example 3 of this invention combined with cupping therapy. Each group consisted of 10 volunteers. The treatment period was 3 months.

[0126] The volunteers met the diagnostic criteria for menopausal syndrome in Traditional Chinese Medicine (TCM). The diagnosis was based on the "Technical Guidelines for Clinical Trials of Traditional Chinese Medicine and Natural Drugs in the Treatment of Menopausal Syndrome in Women" and *Traditional Chinese Medicine Gynecology* (edited by Luo Songping, Higher Education Press, 2008).

[0127] (1) Age: The age of onset is greater than 40 years old.

[0128] (2) Main symptoms: menstrual disorders or hot flashes and sweating during menopause, or mood changes.

[0129] (3) Secondary symptoms: ① lower back pain, dizziness and tinnitus; ② or rib pain, breast tenderness, headache; ③ or palpitations, restlessness, insomnia and dreaminess; ④ or hot palms and soles, dry and burning sensation in the vagina, painful intercourse, dry mouth and constipation; ⑤ or cold pain in the lower back, cold limbs, lethargy, facial and limb edema, decreased libido, clear and copious urine, frequent urination at night, etc.

[0130] (4) The tongue is pale red or reddish, with a thin white or thin yellow coating, and the pulse is thready and rapid or deep and thready.

[0131] A diagnosis can be made if the patient presents with at least one of the following symptoms: (1) and (2) in the disease diagnosis, and / or has at least one of the following secondary symptoms: (1) and (2). The diagnosis can be made by combining the symptoms with the tongue and pulse.

[0132] 2. Treatment Plan

[0133] 2.1 The oral administration group used the tablets of Example 3 of the present invention, 6 tablets each time, 3 times a day; the oral administration + cupping therapy group used the tablets of Example 3 of the present invention, 6 tablets each time, 3 times a day, and cupping therapy was performed twice a week.

[0134] 2.2 Both groups of patients were required to undergo follow-up examinations before treatment and 3 months after treatment. Heart rate variability indicators and TCM pulse diagnosis collected at the initial visit were used as baseline data, while heart rate variability indicators and TCM pulse diagnosis collected 3 months after treatment were used as control data.

[0135] 2.3 Collect, compare, and analyze relevant data to evaluate the therapeutic effects of the two treatment regimens on menopausal syndrome.

[0136] 3. Treatment Results

[0137] The tablets of Example 3 of this invention were used to treat menopausal syndrome. Through the collection, analysis, and comparison of heart rate variability data, traditional Chinese medicine pulse diagnosis, and subjects' physical sensations, the results showed that the subjects experienced varying degrees of improvement in symptoms caused by insufficient Qi and blood production during menopause, including spleen and stomach disharmony, bone and joint pain, memory decline, fatigue, depression, irritability, insomnia, and anxiety. In the oral administration group, 6 subjects experienced symptom disappearance, 3 subjects experienced significant symptom improvement, and 1 subject experienced good symptom improvement. In the oral administration + cupping therapy group, 7 subjects experienced symptom disappearance, 2 subjects experienced significant symptom improvement, and 1 subject experienced good symptom improvement. The overall effective rate reached 100%.

[0138] It is evident that the herbal formula of this invention has the effects of calming the mind and soothing the nerves, strengthening the spleen and stomach, nourishing the liver and tonifying the kidneys. It can not only treat insomnia, but also has a certain effect on improving and treating menopausal syndrome in women. It is also convenient to use, safe, and has significant effects.

[0139] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A traditional Chinese medicine for treating insomnia, characterized in that, The traditional Chinese medicine is made from the following raw materials in parts by weight: 2-8 parts egg yolk, 1-7 parts donkey-hide gelatin, 2-8 parts jujube seed, 1-7 parts Solomon's seal rhizome, 2-8 parts lily bulb, 1-7 parts longan pulp, 2-8 parts ophiopogon root, 2-8 parts angelica root, 1-7 parts clove, 3-10 parts dried tangerine peel, 1-7 parts mulberry leaf, 1-7 parts chrysanthemum, 2-8 parts peach kernel, 4-13 parts kudzu root, 1-7 parts saffron, 2-8 parts ganoderma lucidum, 2-8 parts cistanche deserticola, 2-8 parts raspberry, 2-8 parts wolfberry, 2-8 parts yam, 4-13 parts roasted malt, 1-7 parts roasted ginkgo kernel, 4-13 parts roasted euryale seed, and 2-8 parts prepared licorice root.

2. The traditional Chinese medicine for treating insomnia according to claim 1, characterized in that, The dosage forms of the traditional Chinese medicine include ointments, granules, and tablets.

3. The traditional Chinese medicine for treating insomnia according to claim 2, characterized in that, The preparation method of the ointment includes the following steps: weigh each raw material according to the proportion, wash and dry them, then pulverize them to 100-150 mesh, then mix the obtained powders, add 8-10 times the total weight of pure water of the raw materials, extract for 1-3 hours, filter to obtain extract a and residue a; add 8-10 times the amount of pure water to residue a, extract for 1-3 hours, filter to obtain extract b and residue b; mix extract a and extract b, add 40-60% honey, and concentrate to 1.3 times the concentration to obtain the ointment.

4. The traditional Chinese medicine for treating insomnia according to claim 2, characterized in that, The preparation method of the granules includes the following steps: weigh each raw material according to the proportion, wash and dry them, then pulverize them to 100-150 mesh, then mix the obtained powders, add 8-10 times the total weight of the raw materials in pure water, extract for 1-3 hours, filter to obtain extract a and residue a; add 8-10 times the amount of pure water to residue a, extract for 1-3 hours, filter to obtain extract b and residue b; mix extract a and extract b, concentrate, spray dry to obtain extract powder, then add the extract powder to edible ethanol, mix, dry and granulate in sequence to obtain granules.

5. The traditional Chinese medicine for treating insomnia according to claim 2, characterized in that, The preparation method of the tablets includes the following steps: weigh each raw material according to the proportion, wash and dry them, then pulverize them to 100-150 mesh, then mix the obtained powders, add 8-10 times the total weight of the raw materials in pure water, extract for 1-3 hours, filter to obtain extract a and residue a; add 8-10 times the amount of pure water to residue a, extract for 1-3 hours, filter to obtain extract b and residue b; mix extract a and extract b, concentrate, spray dry to obtain extract powder, then add the extract powder to edible ethanol, mix, dry and granulate in sequence to obtain granules, add excipients to the granules, compress into tablets to obtain tablets.

6. The traditional Chinese medicine for treating insomnia according to claim 5, characterized in that, The excipients include xylooligosaccharides, magnesium stearate, calcium sulfate, sugar powder, lactose, or any combination thereof, and the excipients constitute 0.1-3% of the total weight of the granules.

7. The traditional Chinese medicine for treating insomnia according to any one of claims 3-5, characterized in that, The extraction temperature is 60-90℃.

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