Application of pain and diarrhea prescription in preparation of medicine for preventing and treating ulcerative colitis related colon cancer

Abstract

The invention belongs to the technical field of traditional Chinese medicines, provides an application of a pain and diarrhea essential prescription in preparation of medicines for preventing and treating ulcerative colitis-related colon cancer for the first time, and opens up a new application of the pain and diarrhea essential prescription. A large number of in-vivo and in-vitro experiments prove that the pain and diarrhea essential prescription can inhibit tumor formation by inhibiting tumor cell proliferation and promoting mitochondrial autophagy of colon cancer cells, so that the purpose of preventing and treating the ulcerative colitis related colon cancer is achieved, and the pain and diarrhea essential prescription has a good application prospect in the aspect of preparing the medicine for preventing and treating the ulcerative colitis related colon cancer and has a wide application prospect. The traditional Chinese medicine composition has important significance for treating colon cancer related to colitis.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine, and particularly relates to the application of Tongxie Yaofang in the preparation of medicines for preventing and treating ulcerative colitis-related colon cancer. Background Art

[0002] Ulcerative colitis (UC) is a chronic recurrent intestinal inflammatory disease of unknown cause, which seriously impairs the quality of life of patients. Clinical symptoms are mainly bloody stools with mucus, pus, diarrhea or tenesmus; in acute and critical cases, there are systemic symptoms, and they are often accompanied by extraintestinal diseases and liver damage, arthritis, skin damage, cardiomyopathy, oral ulcers, iridocyclitis and endocrine diseases. At present, the treatment is mainly aimed at the inflammatory response itself. Immunomodulatory drugs such as biological agents induce remission by inhibiting proinflammatory cytokines or blocking the migration of immune cells. However, the current treatment methods are not enough to overcome the heterogeneity of UC, nor can they achieve healing of intestinal epithelial cells (IECs) in a considerable proportion of patients. In recent years, the incidence of UC in my country has increased year by year. The treatment of UC with traditional Chinese medicine can be traced back to the Yellow Emperor's Classic of Internal Medicine more than 2,000 years ago. Traditional Chinese medicine pays more attention to "holistic concept and root cause treatment". This multi-factor and multi-angle understanding of complex diseases is the advanced thinking method of traditional Chinese medicine, which coincides with the modern medicine's greater emphasis on the holistic model.

[0003] Due to its complex etiology, difficulty in curing, frequent recurrences, and close relationship with colon cancer, it has been listed as one of the intractable diseases in modern times by the World Health Organization. The etiology and pathogenesis of UC are still unclear. Western medicine often uses aminosalicylic acid, corticosteroids and immunosuppressants for treatment, which can quickly relieve symptoms, but the drug toxicity and side effects are large; the use of traditional Chinese medicine for individualized syndrome differentiation treatment of UC is reliable and relatively safe, but the problems of chronic persistent attacks and the appearance of many extraintestinal symptoms are still the difficulties in the treatment of UC.

[0004] Through the above analysis, the problems and defects of the prior art are as follows:

[0005] Existing Western medicines for the treatment of ulcerative colitis have significant toxic and side effects, while Chinese medicines are slow to take effect and cause continuous attacks. No drug can achieve the effect of long-term healing of the intestinal mucosal barrier.

[0006] Tongxie Yaofang, which is composed of dried tangerine peel, white atractylodes, white peony root, and saposhnikovia root, is one of the classic prescriptions of traditional Chinese medicine. It was first seen in the Danxi Xinfa written by Zhu Zhenheng in the Yuan Dynasty, and was clearly called "Tongxie Yaofang" in the Jingyue Complete Book of the Ming Dynasty. This prescription is composed of four herbs: stir-fried white atractylodes, stir-fried white peony root, stir-fried dried tangerine peel, and saposhnikovia root. It has the effects of tonifying the spleen and softening the liver, removing dampness and stopping diarrhea. It is mainly used to treat pain and diarrhea caused by liver and spleen disharmony, which manifests as symptoms such as intestinal rumbling, abdominal pain, diarrhea, and pain relief after diarrhea. Modern medical research shows that Tongxie Yaofang has pharmacological effects such as anti-inflammatory, analgesic, and antidiarrheal. It is often used clinically to treat irritable bowel syndrome, colitis, abdominal pain and diarrhea, etc., which are caused by liver hyperactivity and spleen deficiency. Pharmacological experiments have also confirmed the efficacy of Tongxie Yaofang. For example, the research of Dong Guilan et al. showed that Tongxie Yaofang has an inhibitory effect on intestinal motility and analgesic effect on intestinal spasm. It also has antihistamine and antibacterial effects. In conclusion, Tongxie Yaofang, as a traditional Chinese medicine with a long history, has shown its unique therapeutic value in modern medical research and clinical practice, especially in the treatment of digestive system diseases. However, existing studies have not involved the application of Tongxie Yaofang in the treatment of colon cancer. Summary of the invention

[0007] In view of the problems existing in the prior art, the purpose of the present invention is to achieve effective prevention and treatment of ulcerative colitis-related colon cancer, and provide the use of Tongxie Yaofang in the preparation of drugs for preventing and treating ulcerative colitis-related colon cancer.

[0008] The objective of the present invention is achieved through the following technical solutions:

[0009] The present invention provides application of Tongxie Yaofang in preparing medicines for preventing and treating ulcerative colitis-related colon cancer.

[0010] Furthermore, the prescription for treating diarrhea comprises, by weight: 20-30 parts of Atractylodes macrocephala, 15-25 parts of White Peony Root, 10-20 parts of Tangerine Peel, and 10-25 parts of Saposhnikovia divaricata.

[0011] Furthermore, the preparation method of the Tongxie prescription comprises the following steps:

[0012] S101, wash Atractylodes macrocephala, white peony root, dried orange peel, and saposhnikovia divaricata, and soak them in clean water for 10 to 30 minutes respectively;

[0013] S102, mixing the soaked Atractylodes macrocephala, white peony root, dried orange peel and saposhnikovia root together, and adding 1 to 4 times of purified water for decoction;

[0014] S103. When the water boils and only half of it remains, stop boiling; filter and concentrate to obtain the traditional Chinese medicine preparation of Tongxie Yaofang.

[0015] Furthermore, the decoction in step S102 specifically includes:

[0016] 1) Adding medicinal materials: Take the pre-soaked Atractylodes macrocephala, white peony root, dried orange peel, and saposhnikovia root; then take the decoction container, add water to the surface of the medicinal materials, squeeze and soak the medicinal materials until there are no gaps, and add purified water to exceed the upper surface of the medicinal materials;

[0017] 2) First decoction: decoct under a pressure of 0.15 MPa and keep boiling for 10 to 30 minutes; stop heating and collect the liquid medicine while it is hot, squeeze the medicinal materials until there is no residue in the liquid medicine, and store the liquid medicine separately;

[0018] 3) Second decoction: Measure the temperature of the medicinal materials after the first decoction and the squeezed medicinal liquid, and add pure water with a temperature not lower than the measured temperature to the medicinal materials until it exceeds the upper surface of the medicinal materials;

[0019] 4) Second decoction: decoct at normal pressure and maintain boiling, stop heating and collect the medicinal liquid while it is still hot, squeeze the medicinal materials until there is no residual medicinal liquid, combine the two medicinal liquids and mix them evenly.

[0020] Furthermore, the filtering and concentration in step S103 specifically include:

[0021] The water extract after decoction is filtered and separated by an inorganic ceramic membrane separation method, and the obtained filtrate is concentrated into an extract with a relative density of 1.02-1.15 by a nanofiltration membrane with a molecular weight cutoff of more than 150.

[0022] Furthermore, the membrane separation uses an inorganic ceramic membrane with a pore size range of 0.02 to 1 μm; the filtration pressure is 0.01 to 0.8 MPa, the filtration temperature is 4 to 85°C, and the membrane flux is 20 to 300 L / m 2 ·h.

[0023] Furthermore, the medicine is an oral preparation.

[0024] Furthermore, the oral preparation includes any one of ointment, granules, capsules, tablets, powders, and pills.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] The present invention provides for the first time the use of Tongxie Yaofang in the preparation of drugs for preventing and treating ulcerative colitis-related colon cancer, opening up a new use of Tongxie Yaofang. The present invention has verified through a large number of in vivo and in vitro experiments that Tongxie Yaofang can inhibit tumor cell proliferation and promote mitochondrial autophagy of colon cancer cells, thereby inhibiting tumor formation and achieving the purpose of preventing and treating ulcerative colitis-related colon cancer. It has good application prospects in the preparation of drugs for preventing and treating ulcerative colitis-related colon cancer, and is of great significance for solving the difficult problems in the treatment of colitis-related colon cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] The present invention will be further described below in conjunction with the accompanying drawings and embodiments:

[0028] Figure 1 A flow chart of the preparation method of the Tongxie prescription provided in Example 1;

[0029] Figure 2 It was shown that Tongxie Yaofang inhibited the occurrence of colitis-related tumors in the AOM / DSS mouse model (*p<0.05, p<0.01);

[0030] Figure 3 It was shown that the serum containing Tongxie Yaofang significantly inhibited the proliferation of tumor cells (*p<0.05, #p<0.05);

[0031] Figure 4 It was shown that Tongxie Yaofang inhibited tumor formation by promoting mitochondrial autophagy in colon cancer cells (cell experiment) (*P<0.05, **P<0.01, ***P<0.001);

[0032] Figure 5 It was shown that Tongxie Yaofang inhibited tumor formation in vivo by promoting mitochondrial autophagy;

[0033] Figure 6 The component analysis and proteomics of Tongxie Yaofang are shown. DETAILED DESCRIPTION

[0034] The examples are provided to better illustrate the present invention, but the present invention is not limited to the examples. Therefore, those skilled in the art may make non-essential improvements and adjustments to the implementation scheme according to the above invention content, which still fall within the protection scope of the present invention.

[0035] The endpoints and any values ​​of the ranges disclosed in this article are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of each range, the endpoint values ​​of each range and the individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed in this article.

[0036] The present invention will be described in detail below by way of examples. It should be understood that the following examples are only used to further explain and illustrate the content of the present invention in detail, and are not used to limit the present invention.

[0037] It should be noted that the experimental methods described in the following experiments, unless otherwise specified, are all conventional methods, and the reagents and materials, unless otherwise specified, can be obtained from commercial channels.

[0038] The Chinese medicine involved in the following embodiments includes: Atractylodes macrocephala, White Peony Root, Tangerine Peel, Saposhnikovia divaricata, and the pharmacological analysis is as follows:

[0039] 1. Atractylodes macrocephala: Atractylodes macrocephala is mainly used to enhance gastrointestinal function and aid digestion. From a modern medical perspective, compounds in Atractylodes macrocephala can help regulate immune responses and may be beneficial for diseases such as ulcerative colitis that involve immune disorders.

[0040] 2. White Peony Root (Radix Paeoniae Alba): White peony root has analgesic, anti-inflammatory and anti-allergic effects. Such effects are beneficial for ulcerative colitis because one of the main symptoms of this disease is colon inflammation.

[0041] 3. Tangerine peel (Citrus reticulata): Tangerine peel is mainly used to regulate gastrointestinal function and help digestion. It helps relieve digestive discomfort such as flatulence and cramps.

[0042] 4. Radix Saposhnikoviae: Radix Saposhnikoviae is mainly used to relieve exterior symptoms in traditional Chinese medicine. It also has anti-inflammatory and analgesic effects. It may be helpful for ulcerative colitis and the pain and inflammation it causes.

[0043] Embodiment 1:

[0044] This embodiment provides the use of Tongxie Yaofang in the preparation of drugs for preventing and treating ulcerative colitis-related colon cancer. The specific composition of the Tongxie Yaofang is: 30g of Atractylodes macrocephala, 20g of White Peony Root, 15g of Tangerine Peel, and 10g of Saposhnikovia divaricata. Figure 1 The preparation method shown is:

[0045] S101. Use clean water to quickly rinse Atractylodes macrocephala, white peony root, dried tangerine peel, and Saposhnikovia divaricata once or twice, and soak them in clean water for 30 minutes respectively. Be careful not to soak them for too long.

[0046] S102, mixing the soaked Atractylodes macrocephala, white peony root, dried orange peel and saposhnikovia root together, adding 3 times of purified water for decoction; the specific steps are as follows:

[0047] 1) Adding medicinal materials: Take the pre-soaked Atractylodes macrocephala, white peony root, dried orange peel, and saposhnikovia root; then take the decoction container, add water to the surface of the medicinal materials, squeeze and soak the medicinal materials until there are no gaps, and add purified water to exceed the upper surface of the medicinal materials;

[0048] 2) First decoction: decoct under a pressure of 0.15 MPa and keep boiling for 30 minutes; stop heating and collect the liquid medicine while it is hot, squeeze the medicinal materials until there is no residue in the liquid medicine, and store the liquid medicine separately;

[0049] 3) Second decoction: Measure the temperature of the medicinal materials after the first decoction and the squeezed medicinal liquid, and add pure water with a temperature not lower than the measured temperature to the medicinal materials until it exceeds the upper surface of the medicinal materials by 2 cm;

[0050] 4) Second decoction: decoct at normal pressure and maintain boiling for 30 minutes, stop heating and collect the medicinal liquid while it is still hot, squeeze the medicinal materials until there is no residual medicinal liquid, combine the two medicinal liquids and mix them evenly.

[0051] S103. Filter and concentrate the water extract obtained by the above decoction to obtain a traditional Chinese medicine preparation for treating ulcerative colitis-related colon cancer.

[0052] The filtering and concentration specifically include:

[0053] The water extract after decoction is filtered and separated by an inorganic ceramic membrane separation method, and the obtained filtrate is concentrated into an extract with a relative density of 1.02-1.15 by a nanofiltration membrane with a molecular weight cutoff of more than 150.

[0054] Furthermore, the membrane separation uses an inorganic ceramic membrane with a pore size range of 0.02 to 1 μm; the filtration pressure is 0.01 to 0.8 MPa, the filtration temperature is 4 to 85°C, and the membrane flux is 20 to 300 L / m 2 ·h.

[0055] Test case: The therapeutic effect of Tongxie Yaofang on ulcerative colitis-related colon cancer

[0056] In order to explore the therapeutic effect of the Tongxie prescription described in this example on colitis, animal experiments and cell experiments were conducted respectively. The specific contents of the experiments are as follows:

[0057] Experimental Animals:

[0058] Male C57BL / 6, 6–8 weeks old, 20–22 g, a total of 30 animals were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd. (Jinan, China) (Experimental Animal License No.: SCXK(Lu)20200018).

[0059] Mice were housed under specific pathogen-free conditions. The ambient temperature was maintained at 21 ± 2 °C with 12 hours of light. Mice had free access to food and water. All steps were performed in accordance with the guidance of the Animal Research Ethics Committee of the Affiliated Hospital of Qingdao University and in accordance with the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health (No. AHQU-MAL20221014; No. AHQU-MAL20230217). After 1 week of adaptive feeding, the CAC mouse model (colitis-associated colon cancer mouse model) was established with reference to existing methods. The specific method was as follows: azomethane (AOM, Sigma Aldrich, USA) was injected intraperitoneally into the model mice once. After 3 days, 3% dextran sulfate (DSS, Meilunbio, China) water was drunk for 5 days, and then water was drunk for 15 days. The cycle of DSS and normal water was repeated 3 times. Mice were evenly allocated according to the random number table method (n = 6 per group). The treatment of model mice is as follows:

[0060] The low-dose group, medium-dose group, and high-dose Tongxie Yaofang group (i.e., TXYF group) were given 4.9, 9.75, and 19.5 g / (kg·d) TXYF by gavage daily, respectively. The control group and the model group were given the same volume of water by gavage. The dosage was determined based on the dose-animal conversion between humans and animals. The mice were treated in groups throughout the experiment. The mice were euthanized by carbon dioxide inhalation on the 62nd day of the experiment, and the colons were collected.

[0061] like Figure 2 As shown in the animal experiment, it was found that the treatment of Tongxie Yaofang with AOM / DSS modeling could significantly reduce the tumor formation rate, improve the pathological score, and increase the survival rate. Specifically, in the DSS / AOM-induced colitis model of mice, it was found that low, medium, and high doses of Tongxie Yaofang could inhibit the formation of colitis-related tumors and increase the survival rate ( Figure 2 Tongxie Yaofang improved the weight loss and mortality of UC-CRC mice ( Figure 2 B, C). HE of colon in the control group, model group, and Tongxie Yaofang group. The model group showed extensive and continuous adenocarcinoma tissue, while the Tongxie Yaofang group showed focal adenoma ( Figure 2 (EG).

[0062] Cell experiments:

[0063] Male Sprague-Dawley rats (7-8 weeks old, 300-320 g) were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd. (Jinan, China) with license number SYXK(Lu)20190003. Twenty rats were randomly divided into two groups. The TXYF group was given 6.70 g / (kg·d) by gavage twice a day for 5 days, and the blank control group was given an equal volume of water. One hour after the last administration, the rats were anesthetized with isoflurane. Abdominal aortic blood was collected from all rats. Rats were euthanized by carbon dioxide inhalation. Serum was separated, inactivated at 56°C, sterile filtered with a 0.22 μm microporous membrane filter, and stored at -80°C to obtain the drug-containing serum of Tongxie Yaofang.

[0064] Cell culture: Human colon cancer cells HT-29 were purchased from Pricella (China). Human normal colon epithelial cells NCM460 were purchased from the American Culture Collection (USA). HT-29 and NCM460 cells were cultured in Dulbecco's Modified Eagle's Medium / Nutrient Mixture F-12 (Gibco, USA) containing 10% fetal bovine serum (Gibco, USA) and 1% penicillin-streptomycin (Meilune, China) in an incubator at 37°C and 5% carbon dioxide. Lipopolysaccharide (LPS, 1 μg / mL, MedChemExpress, USA) was used to treat HT-29 and NCM460 cells for 48 hours to induce inflammation, and the HT-29 inflammatory tumor model was obtained.

[0065] like Figure 3 As shown in the cell experiment, it was found that the serum containing Tongxie Yaofang significantly inhibited the proliferation of tumor cells and promoted mitochondrial autophagy. Specifically, 5%, 10%, and 20% of the serum containing Tongxie Yaofang were applied to the LPS-induced HT-29 inflammatory tumor model. When the concentration was ≥10%, the cell viability was inhibited ( Figure 3 A). 10% Tongxie Yaofang can significantly inhibit the levels of inflammatory factors TNF-α and IL-6 in HT-29 cells induced by LPS after 12 hours ( Figure 3 (B, C). 10% Tongxie Yaofang intervention for 12 days can significantly inhibit LPS-induced HT-29 cell colony formation ( Figure 3 (D, E). 10% Tongxie Yaofang intervened in LPS-induced HT-29 cells for 24 h and promoted the expression of mitochondrial autophagy-related proteins ( Figure 3 (F, G).

[0066] Furthermore, in order to explore the mechanism of Tongxie Yaofang in inhibiting colon cancer, the following experiments were conducted:

[0067] Knockdown of PINK1: Small interfering RNA targeting human PINK1 mRNA was synthesized by Servicebio (China). The sequences were: forward 5'-GCCAGUACCUUUGUGUGAATT-3' and reverse 5'-UUCACACAA-AGGUACUGGCTT-3'. PINK siRNA was transfected into HT-29 cells using Lipofectamine 2000 (Invitrogen, USA) according to the manufacturer's instructions.

[0068] Western blot: Total proteins were extracted from tissues and cells using RIPA buffer (Boster, China) supplemented with 1% protease inhibitors (Boster, China). Mitochondrial proteins were isolated from cells according to the instructions of the mitochondrial isolation kit (Sangon Biotech, China). Protein concentration was measured using a BCA kit (Beyotime Biotechnology, China). Proteins were then separated using SDS-PAGE gel (Servicebio, China) and transferred to polyvinylidene difluoride membranes (Millipore, Germany), followed by incubation with primary antibodies at 4°C overnight. The protein bands were incubated with the corresponding HRP-labeled antibodies. The results were detected using a chemiluminescent detection reagent (Servicebio, China) according to the manufacturer's instructions. β-actin was used as a total protein control and TOM20 as a mitochondrial protein control.

[0069] like Figure 4 As shown, Tongxie Yaofang inhibits tumor formation by promoting mitochondrial autophagy in colon cancer cells.

[0070] HT-29 cells were treated with lysosomal inhibitor chloroquine CQ and PINK1 siRNA to observe the role of TXYF in mitosis. These results indicate that TXYF inhibits EMT and activates PTEN expression through autophagy. TXYF promotes the expression of LC3II in CQ-treated HT-29 cells ( Figure 4 Middle A). PINK1 knockout inhibited the inhibitory effect of TXYF on cell EMT and the promoting effect of TXYF on PTEN ( Figure 4 These results suggest that TXYF inhibits the progression of CAC by activating the PINK1 / Parkin pathway in mitophagy.

[0071] like Figure 5 As shown, Tongxie Yaofang inhibits tumor formation in vivo by promoting mitochondrial autophagy.

[0072] The survival rate of mice treated with TXYF and CQ was lower than that of the TXYF group, but there was no statistically significant difference among the groups ( Figure 5 Middle A). Late TXYF treatment had a protective effect on the body weight of mice, while CQ attenuated this effect ( Figure 5 (A). CQ had a protective effect on the body weight of mice, while CQ weakened this effect ( Figure 5 Middle B). The colon of each group of mice ( Figure 5 CQ significantly inhibited TXYF-induced mitophagy. In the CAC mouse model, the improvement effect of TXYF on EMT and PTEN was inhibited after CQ treatment ( Figure 5These results support that TXYF inhibits tumor progression in CAC mice via mitophagy.

[0073] like Figure 6 As shown, the component analysis and proteomics of Tongxie Yaofang.

[0074] UPLC-MS / MS analysis detected the composition of TXYF ( Figure 6 To explore the potential mechanism of TXYF treatment, proteomics was used to detect protein changes. In LPS-induced NCM460 cells, 42 proteins were increased and 83 proteins were decreased in NCM460 cells treated with 10% TXYF compared with control cells ( Figure 6 (B). Subcellular localization prediction analysis showed that the differentially expressed proteins were mainly located in the cell membrane. The expressed proteins were mainly located in the cytoplasm, nucleus, mitochondria and extracellular matrix ( Figure 6 Middle C). Proteins related to tumor biology, mitochondrial function, and autophagy ( Figure 6 (in DF).

[0075] Finally, it should be noted that the above is only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred arrangement scheme, a person skilled in the art should understand that the technical solution of the present invention may be modified or replaced by equivalents without departing from the spirit and scope of the technical solution of the present invention.

Claims

1. Application of Tongxie Yaofang in the preparation of drugs for the prevention and treatment of ulcerative colitis-related colon cancer.

2. The use according to claim 1, characterized in that: The prescription for treating diarrhea includes, by weight: 20-30 parts of Atractylodes macrocephala, 15-25 parts of White Peony Root, 10-20 parts of Tangerine Peel, and 10-25 parts of Saposhnikovia divaricata.

3. The use according to claim 1 or 2, characterized in that: The preparation method of the Tongxie prescription comprises the following steps: S101, wash Atractylodes macrocephala, white peony root, dried orange peel, and saposhnikovia divaricata, and soak them in clean water for 10 to 30 minutes respectively; S102, mixing the soaked Atractylodes macrocephala, white peony root, dried orange peel and saposhnikovia root together, and adding 1 to 4 times of purified water for decoction; S103. When the water boils and only half of it remains, stop boiling; filter and concentrate to obtain the traditional Chinese medicine preparation of Tongxie Yaofang.

4. The use according to claim 3, characterized in that: The decoction in step S102 specifically includes: 1) Adding medicinal materials: Take the pre-soaked Atractylodes macrocephala, white peony root, dried orange peel, and saposhnikovia root; then take the decoction container, add water to the surface of the medicinal materials, squeeze and soak the medicinal materials until there are no gaps, and add purified water to exceed the upper surface of the medicinal materials; 2) First decoction: decoct under a pressure of 0.15 MPa and keep boiling for 10 to 30 minutes; stop heating and collect the liquid medicine while it is hot, squeeze the medicinal materials until there is no residue in the liquid medicine, and store the liquid medicine separately; 3) Second decoction: Measure the temperature of the medicinal materials after the first decoction and the squeezed medicinal liquid, and add pure water with a temperature not lower than the measured temperature to the medicinal materials until it exceeds the upper surface of the medicinal materials; 4) Second decoction: decoct at normal pressure and maintain boiling, stop heating and collect the medicinal liquid while it is still hot, squeeze the medicinal materials until there is no residual medicinal liquid, combine the two medicinal liquids and mix them evenly.

5. The use according to claim 3, characterized in that: The filtering and concentration in step S103 specifically include: The water extract after decoction is filtered and separated by an inorganic ceramic membrane separation method, and the obtained filtrate is concentrated into an extract with a relative density of 1.02-1.15 by a nanofiltration membrane with a molecular weight cutoff of more than 150.

6. The use according to claim 5, characterized in that: Membrane separation uses inorganic ceramic membranes with a pore size range of 0.02 to 1 μm; the filtration pressure is 0.01 to 0.8 MPa, the filtration temperature is 4 to 85 °C, and the membrane flux is 20 to 300 L / m 2 ·h.

7. The use according to claim 1, characterized in that: The medicine is an oral preparation.

8. The use according to claim 7, characterized in that: The oral preparation includes any one of ointment, granules, capsules, tablets, powders, and pills.

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