Rheum officinale and herba epimedii medicine pair combined medicine as well as preparation method and application thereof

Abstract

The invention relates to the technical field of traditional Chinese medicine, in particular to a rhubarb and herba epimedii medicine pair combined medicine, a preparation method and application of the rhubarb and herba epimedii medicine pair combined medicine in preparation of medicine for delaying and / or treating organ aging. The combined medicine of herba epimedii and radix et rhizoma rhei is adopted, the good activity is achieved in the anti-aging related diseases, compared with a proved recipe, the composition is simple, and more importantly, the activity of a product obtained through mixed enzyme assisted ethanol reflux extraction of the two medicine pairs is obviously superior to that of a product obtained through other conventional extraction methods; the extract is superior to a single rheum officinale extract and a single herba epimedii extract. The traditional Chinese medicine components, the weight ratio of the traditional Chinese medicine components and the extraction process are optimized, synergistic effects are achieved, and the traditional Chinese medicine composition has the advantage of being good in curative effect on visceral organ aging diseases and has good application prospects.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine, and specifically, it is a combination drug of rhubarb and epimedium, a preparation method thereof, and an application thereof. Background Art

[0002] Rhubarb is the root and rhizome of Rheum palmatum L., Rheum tanguticum Maxim. ex Balf., or Rheum officinale Baill. of the Polygonaceae family. The 2020 edition of the pharmacopoeia states that it has the effects of purging and attacking accumulation, clearing heat and purging fire, cooling blood and detoxifying, removing stasis and dredging meridians, promoting diuresis and excreting jaundice, etc., and is widely used clinically for the treatment of diseases such as defecation, pancreatitis, and atherosclerosis; its active ingredients are mainly anthraquinone compounds, anthrones, and stilbenes. Anthraquinone compounds have various biological activities, such as anti-inflammatory, antioxidant, anti-apoptotic, and tissue damage repair effects. In recent years, reports have pointed out the therapeutic potential of rhubarb and its derivatives in chronic nephritis.

[0003] Epimedium is a traditional tonifying Chinese medicine, which is the dried leaf of Epimedium brevicornu Maxim., Epimedium sagittatum (Sieb.) Maxim., Epimedium pubescens Maxim., or Epimedium koreanum Nakai of the Berberidaceae family, and has the functions of tonifying kidney yang, strengthening tendons and bones, and dispelling wind and dampness. Modern research has confirmed that it can be used for pharmacological effects such as anti-aging, anti-osteoporosis, and anti-tumor, and is clinically mostly used for the treatment of male infertility, osteoporosis, chronic hepatitis, bronchitis, and cardiovascular diseases. Experiments have shown that epimedium can increase the testosterone level in subacutely aged male rats induced by D-galactose. In addition, after icariin intervention, the serum cortisol content in kidney-yang deficiency rats can be increased, and it has an inhibitory effect on the decrease in serum testosterone content in kidney-yang deficiency mice.

[0004] However, there is currently no report on the combination drug of rhubarb and epimedium and its application in delaying organ aging. Summary of the Invention

[0005] The purpose of the present invention is to provide a combination drug of rhubarb and epimedium, a preparation method thereof, and an application thereof in the preparation of a drug for delaying organ aging.

[0006] Research has proven that rhubarb is bitter in taste and cold in nature. It mainly enters the spleen, stomach, large intestine, liver, and pericardium meridians, and functions to purge fire and promote defecation, cool blood and detoxify, and remove stasis and dredge channels. It is commonly used for symptoms such as stagnant constipation, jaundice, and stranguria. Modern pharmacological research shows that rhubarb has effects such as anti-inflammatory, antiviral, antioxidant, and lipid-regulating. Epimedium, also known as Xianlingpi, is warm in nature, pungent and sweet in taste, and enters the liver and kidney meridians. It has the effects of tonifying the kidney yang, strengthening the bones and muscles, and dispelling wind-dampness. Epimedium and its main active ingredients have effects such as anti-aging and anti-inflammatory. In order to improve the impact of long-term use of rhubarb on gastrointestinal function, and to solve the problems commonly existing in current traditional Chinese medicine compositions, such as a large variety of formula herbs, low content of active ingredients, and poor bioavailability. Based on the activities of rhubarb and epimedium, the present invention provides a combined drug of rhubarb and epimedium medicine pair, a preparation method thereof, and its application in the preparation of drugs for delaying organ aging.

[0007] In the first aspect of the present invention, there is provided a medicine pair composition composed of rhubarb and epimedium, and the weight ratio of the rhubarb and epimedium is 1:1 - 1:10.

[0008] In the second aspect of the present invention, there is provided an application of the medicine pair composition as described above in the preparation of drugs for delaying and / or treating organ aging.

[0009] Furthermore, the organ is the kidney, liver, or spleen.

[0010] In the third aspect of the present invention, there is provided an extract of the medicine pair composition, and the extract is an extract of a mixture of rhubarb and epimedium herbs, and the weight ratio of the rhubarb and epimedium is 1:1 - 1:10.

[0011] Furthermore, the weight ratio of the rhubarb and epimedium is 2:7.

[0012] Furthermore, the extract is an extract obtained by water decoction method or 75% ethanol reflux extraction of a mixture of rhubarb and epimedium herbs.

[0013] Even further, the method of 75% ethanol reflux extraction is as follows: The mixture of rhubarb and epimedium herbs in proportion is crushed and mixed, and 75% ethanol 14 times the weight of the herbs is used as the lysosome, reflux extracted 3 - 4 times, 40 minutes each time, to obtain an extraction solution of the combined drug of rhubarb and epimedium medicine pair, and concentrated under reduced pressure into a paste.

[0014] Further, the extract is obtained by extracting a mixture of rhubarb and epimedium herbs using a mixed enzyme-assisted 75% ethanol reflux method. The extraction method is as follows: Mix the rhubarb and epimedium herb mixture in proportion, crush and mix them, first perform enzymatic pretreatment. The enzymatic hydrolysis conditions are 0.8% (based on the mass of the extract) of an enzyme mixture (cellulase: pectinase: flavonoid glycosidase = 2:2:1 mass ratio), the enzymatic hydrolysis temperature is 40 °C, the initial pH value of the enzymatic hydrolysis solution is 5.0, and the enzymatic hydrolysis is carried out for 2.5 h; then use 75% ethanol as the solvent for ethanol reflux extraction to prepare the extract of the rhubarb and epimedium herb pair composition.

[0015] Further, the weight ratio of the rhubarb and epimedium herbs is 1:1 - 1:10.

[0016] Furthermore, the weight ratio of the rhubarb and epimedium herbs is 2:7.

[0017] In the fourth aspect of the present invention, there is provided an application of the extract of the herb pair composition as described above in the preparation of a drug for delaying and / or treating visceral aging.

[0018] Further, the viscera are the kidney, liver or spleen.

[0019] The advantages of the present invention are as follows:

[0020] 1. The present invention uses a combined drug of epimedium and rhubarb, which exhibits good activity in anti-aging related diseases. Compared with traditional prescriptions and empirical formulas with simple ingredients, more importantly, the product obtained by extracting the two herb pairs using a mixed enzyme-assisted ethanol reflux method has significantly better activity than other conventional extraction methods, as well as better than the single herb extract of rhubarb and the single herb extract of epimedium.

[0021] 2. The present invention optimizes the Chinese medicine ingredients, their weight ratio, and the extraction process, achieving a synergistic effect, having the advantage of good curative effect in treating visceral aging diseases, and having good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 . Effects of the herb pair composition on the staining characteristics of renal aging in 5 / 6 (A / I) rats.

[0023] Figure 2 . Effects of the experimental group Ⅳ of the herb pair composition on the staining characteristics of renal, hepatic and splenic tissue aging in the D-gal-induced aging mouse model. DETAILED DESCRIPTION OF THE INVENTION

[0024] The following provides a detailed description of the specific embodiments of the present invention in conjunction with the examples.

[0025] Example 1: Investigation of the preparation method of the rhubarb-epimedium herb pair combined drug

[0026] Rhubarb and epimedium were crushed into coarse powder, divided into 4 groups, and extracted by conventional water decoction, reflux extraction with 75% ethanol as the solvent, and mixed enzyme-assisted reflux extraction with 75% ethanol respectively.

[0027] Water decoction method: Based on the water decoction process of rhubarb and epimedium, it was improved. Specifically, after rhubarb and epimedium were ground into coarse powder, they were decocted with 14 times the weight of the medicinal materials in water for 45 minutes. During the decoction process, it was stirred at any time, filtered while it was hot with two layers of gauze, the residue was washed with a small amount of water, and the medicinal residue was squeezed into a paste by hand.

[0028] Reflux extraction method: 75% ethanol with 14 times the weight of the medicinal materials was used as the solvent to reflux extract the rhubarb and epimedium composition, and the extraction was carried out 3 times, each time for 45 minutes.

[0029] Mixed enzyme-assisted 75% ethanol reflux method: First, enzymatic hydrolysis pretreatment was carried out. The enzymatic hydrolysis conditions were 0.8% (calculated based on the volume of the extract solution) of the enzyme mixture (cellulase: pectinase: flavonoid glycosidase = 2:2:1 mass ratio), the enzymatic hydrolysis temperature was 40 °C, the initial pH value of the enzymatic hydrolysis solution was 5.0, and the enzymatic hydrolysis was carried out for 2.5 hours. Then, ethanol reflux extraction was carried out according to the above reflux method to prepare the extract of the rhubarb and epimedium medicine pair composition, and it was concentrated under reduced pressure into a paste.

[0030] The water extraction group, 75% ethanol reflux extraction group, and enzyme-assisted 75% ethanol reflux extraction group were prepared.

[0031] Table 1 Weighed 3 groups of rhubarb and epimedium medicinal materials with a mass ratio of 1:1, and the extraction rates of rhein, emodin, and icariin were analyzed by the extraction processes of 3 methods respectively. The sum of the index contents was used as the total extraction amount and used as the evaluation index to obtain the results of the extraction process verification test. The results showed that the enzyme-assisted 75% ethanol reflux extraction group had the best extraction effect, so enzyme-assisted 75% ethanol reflux extraction was selected.

[0032] Table 1 Effects of different extraction methods on the contents of the main components of the medicine pair composition (mg / g)

[0033]

[0034] Example 2: Investigation on the active effects of the combined drug of the rhubarb and epimedium medicine pair

[0035] Weigh 20 g each of rhubarb and epimedium, and grind them into coarse powder. The optimal extraction method in Example 1 (enzymatic-assisted reflux extraction with 75% ethanol): First, perform enzymatic hydrolysis pretreatment. The enzymatic hydrolysis conditions are 0.8% (calculated based on the volume of the extract solution) of an enzyme mixture (cellulase:pectinase:flavonoid glycosidase = 2:2:1 mass ratio), the enzymatic hydrolysis temperature is 40 °C, the initial pH value of the enzymatic hydrolysis solution is 5.0, and the enzymatic hydrolysis is carried out for 2.5 h. Then, use 75% ethanol, which is 14 times the weight of the medicinal materials, as the lysing enzyme for reflux extraction. Extract 3 times, each time for 45 min, to prepare the experimental group I of the rhubarb-epimedium combined drug.

[0036] Weigh 20 g of rhubarb and 100 g of epimedium, and grind them into coarse powder. Prepare the experimental group II of the rhubarb-epimedium combined drug according to the above method.

[0037] Weigh 20 g of rhubarb and 200 g of epimedium, and grind them into coarse powder. Prepare the experimental group III of the rhubarb-epimedium combined drug according to the above method.

[0038] Weigh 400 g of rhubarb and 1.4 kg of epimedium, and grind them into coarse powder. Prepare the experimental group IV of the rhubarb-epimedium combined drug according to the above method.

[0039] Weigh 200 g of rhubarb, and grind it into coarse powder. Prepare the rhubarb control group according to the above method.

[0040] Weigh 200 g of epimedium, and grind it into coarse powder. Prepare the epimedium control group according to the above method.

[0041] Select SPF-grade rats, 90 males. Except for the negative control group, the remaining rats were used to construct a CKD model by 5 / 6 (A / I) nephrectomy using the conventional surgical method. Then, the normal group rats and the model group rats were intragastrically administered with normal saline at a dose of 10 ml / kg. The daily dosage of the experimental groups I, II, III, IV, rhubarb, and epimedium groups was intragastrically administered according to 3 g / kg of the original medicinal materials -1 body weight, and losartan potassium (Losartan, Los) was 33.3 mg·kg -1 per day for 30 d. 24 h after the last administration, all rats were anesthetized and sacrificed by collecting blood from the abdominal aorta, and the kidney tissues were collected for β-galactosidase (SA-β-gal) staining.

[0042] The SA-β-gal detection results showed that after the 5 / 6 (A / I) nephrectomy model was established, the blue-green positive staining in the rat kidney tissue increased. After the intervention with the medicinal pair composition, the positive area of senescence staining was significantly reduced, as Figure 1 shown.

[0043] Figure 1 This is the effect of the medicinal pair composition on the senescence staining characteristics of the kidneys of 5 / 6 (A / I) rats. As Figure 1As shown in the figure, senescence occurred in the kidneys of 5 / 6 (A / I) rats, with an increase in the positive staining of SA-β-gal. The combination of rhubarb and epimedium could significantly reduce the positive expression of SA-β-gal staining in the renal tissues of rats in the experimental group. In the comparison, it was found that the activity of the combination of rhubarb and epimedium was significantly better than that of the single rhubarb group and the single epimedium group. In summary, the combination of rhubarb and epimedium had a significant pair effect, and the ethanol extracts of the combination of rhubarb and epimedium in each group had obvious anti-aging pharmacological effects and had a definite therapeutic effect on renal senescence.

[0044] Among them, the combination of rhubarb and epimedium herbs (weight ratio 2:7) had the best effect, followed by the two combinations of rhubarb and epimedium herbs (weight ratio 1:10) and rhubarb and epimedium herbs (weight ratio 1:5), and the combination of rhubarb and epimedium herbs (weight ratio 1:1) had the worst effect. From the above, it can be known that the weight ratio of rhubarb to epimedium herbs of 2:7 is the best ratio.

[0045] Example 3: Investigation on the effect of the combination of rhubarb and epimedium on organ senescence in experimental group Ⅳ

[0046] Twenty-four male C57BL / 6 mice were selected. Except for the negative control group, the remaining mice were intraperitoneally injected with D-gal (200 mg·kg -1 ·d -1 ) to establish a senescence model. Then, the normal group of rats and the model group of mice were intragastrically administered with normal saline at a dose of 20 ml / kg. The daily dosage of experimental group Ⅳ was intragastrically administered according to the original herbs at 5 g / kg -1 body weight. The VE group was intragastrically administered with vitamin E at 100 mg / kg for 30 days. Twenty-four hours after the last administration, all the mice were anesthetized and sacrificed by collecting blood from the abdominal aorta. The kidney, liver, and spleen tissues of the mice were collected for β-galactosidase (SA-β-gal) staining.

[0047] The SA-β-gal detection results showed that after the establishment of the D-gal senescence model, the blue-green positive staining in different tissues of the mice increased. After the intervention with the combination of herbs, the positive area of senescence staining was significantly reduced, as Figure 2 shown.

[0048] Figure 2 Figure is the effect of experimental group Ⅳ of the combination of rhubarb and epimedium on the senescence staining characteristics of the kidney, liver, and spleen tissues of D-gal senescent mice. As Figure 2 shown, compared with the blank group, the positive area of SA-β-gal staining in the kidney, liver, and spleen tissues of the mouse model group increased significantly; compared with the model group, each drug treatment group could reduce the positive expression of SA-β-gal staining in the kidney, liver, and spleen, and experimental group Ⅳ was significantly better than the VE group.

[0049] The present invention optimizes the traditional Chinese medicine components, the weight ratio thereof and the extraction process, and has the advantages of synergistic effect and good curative effect in treating visceral aging diseases, and has good application prospects.

[0050] The preferred embodiments of the present invention have been specifically described above, but the present invention is not limited to the described embodiments. Those skilled in the art can also make various equivalent modifications or substitutions without departing from the spirit of the present invention, and these equivalent modifications or substitutions are all included within the scope defined by the claims of this application.

Claims

1. A pharmaceutical composition, characterized in that, It is composed of rhubarb and epimedium, and the weight ratio of rhubarb to epimedium is 1:1 - 1:

10.

2. The pharmaceutical pair composition according to claim 1, wherein The weight ratio of rhubarb to epimedium is 2:

7.

3. Use of a pair of medicinal compositions as described in claim 1 or 2 in the preparation of a drug for delaying and / or treating visceral aging.

4. A pharmaceutical composition extract, characterized in that, The extract is an extract of a medicinal material mixture of rhubarb and epimedium, and the weight ratio of rhubarb to epimedium is 1:1 - 1:

10.

5. The extract of the pair of medicinal compositions according to claim 4, characterized in that, The weight ratio of rhubarb to epimedium is 2:

7.

6. The extract of the medicine pair composition according to claim 4, characterized in that, The extract is an extract obtained by water decoction method or 75% ethanol reflux extraction from a medicinal material mixture of rhubarb and epimedium.

7. The extract of the pair of medicinal compositions according to claim 5, characterized in that, The method of 75% ethanol reflux extraction is as follows: Mix the mixture of rhubarb and epimedium medicinal materials in proportion, crush and mix them, use 14 times the weight of the medicinal materials of 75% ethanol as the solvent, reflux extract 3 - 4 times, 40 minutes each time, to obtain an extract of the rhubarb and epimedium pair combination drug.

8. The extract of the pair of medicinal compositions according to claim 4, characterized in that The extract is an extract obtained by using a mixed enzyme - assisted 75% ethanol reflux method for extracting a medicinal material mixture of rhubarb and epimedium. The extraction method is as follows: Mix the mixture of rhubarb and epimedium medicinal materials in proportion, crush and mix them, first perform enzymatic hydrolysis pretreatment. The enzymatic hydrolysis conditions are adding an enzyme mixture accounting for 0.8% of the extract quality. The enzyme mixture is a mixture of cellulase, pectinase and flavonoid glycosidase with a mass ratio of 2:2:

1. The enzymatic hydrolysis temperature is 40 °C, the initial pH value of the enzymatic hydrolysis solution is 5.0, and the enzymatic hydrolysis time is 2.5 h; then use 75% ethanol as the solvent for ethanol reflux extraction to prepare an extract of the rhubarb and epimedium pair composition.

9. Use of an extract of a pair of medicinal compositions as described in any one of claims 4 - 8 in the preparation of a drug for delaying and / or treating visceral aging.

10. The application according to claim 3 or 9, characterized in that, The viscera are the kidney, liver or spleen.

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