TW202135848A - Traditional chinese medicine formula flora capsule, preparation method and use thereof in preparing medicine for treating type 2 diabetes - Google Patents

Abstract

The invention belongs to the field of medicines, and particularly relates to a traditional Chinese medicine formula flora capsule which is prepared by matching freeze-dried powder separated from fecal floral and traditional Chinese medicine extraction powder obtained by crushing traditional Chinese medicine extraction effective ingredients. The invention combines the traditional Chinese medicine extraction method and deeply integrates it with the fecal floral separation process to prepare the traditional Chinese medicine formula flora capsule, which greatly improves the treatment effect while changing the administration method, and realizes an efficient and comfortable treatment way.

Description

Chinese medicine formula bacterial group capsule, preparation method and application in preparation of medicine for treating type 2 diabetes

The invention belongs to the technical field of medicine, and specifically relates to the technical field of fecal bacteria transplantation, and more specifically to a traditional Chinese medicine formula fecal bacteria capsule, a preparation method thereof, and an application in the preparation of a medicine for treating type 2 diabetes.

Changes in the structure and function of the intestinal flora can cause a series of metabolic diseases such as obesity and type 2 diabetes. Recent studies have suggested that fecal bacteria transplantation can improve the level of blood sugar control in type 2 diabetes, improve metabolism, and increase insulin sensitivity.

"Fecal microbiota transplantation" (FMT) is defined as transplanting the functional flora in the stool of a healthy person into the gastrointestinal tract of the patient through nasogastric or duodenal tube, gastroscope or colonoscopy, rectal catheter enema, etc. , Rebuild the balance of the intestinal flora, repair the barrier of the intestinal mucosa, and realize the treatment of specific intestinal and extraintestinal diseases.

However, although the above-mentioned administration methods can achieve the purpose of intestinal bacteria transplantation, they need to be infused into the patient’s intestinal tract under a tube state or endoscopy. The patient’s physiological and psychological acceptance is limited, which limits this The promotion and application of treatment techniques in clinical practice. For example, Tian Hongliang et al. (Analysis of the clinical efficacy of bacteria capsule in the treatment of 15 cases of chronic transmission constipation, Chinese Journal of Practical Surgery, Volume 36, Issue 4, April 2016) reported that the use of bacteria capsule in the treatment of 15 cases of chronic transmission constipation was analyzed. The results of the study are preliminary attempts and were not included in the control group. How to overcome the shortcomings of fecal bacteria transplantation is still not enough. In particular, the content of the bacterial capsule in this article is a single bacterial liquid, and the addition of conventional auxiliary materials will reduce the bacterial liquid content. Therefore, it is necessary to conduct further research and development to better apply fecal bacteria transplantation to the treatment of diseases.

In view of the above-mentioned problems, the inventors used the traditional method of administration to extract fresh bacterial liquid to prepare freeze-dried powder. However, the pure bacterial liquid freeze-dried powder sometimes requires the use of Chinese medicine to improve the quality of treatment. The inventor combined the traditional Chinese medicine extraction method to deeply integrate the two processes to form a traditional Chinese medicine formula flora capsule, which greatly improves the treatment effect while changing the administration method, and realizes an efficient and comfortable treatment route.

The technical solution provided by the present invention is:

On the one hand, the present invention provides a traditional Chinese medicine formula flora capsule, which is characterized in that it is prepared by pulverizing the lyophilized powder after the separation of fecal bacteria and the traditional Chinese medicine extraction active ingredients to obtain the traditional Chinese medicine extract powder.

In a specific embodiment, the method for separating the fecal bacteria is the separation of the microorganisms in the cavity contents by the separation device, more specifically, the separation is performed by the device described in the 201410606325.4 patent.

More specifically, the preparation method of the freeze-dried powder after fecal bacteria separation includes the following steps:

(1) Use a sterile sampling bottle to collect stool samples from healthy donors, seal them and send them to the sterile operating room, and place them in series with the filter bottles, AB bottles, deodorizing bottles, buffer bottles, and bacteria liquid bottles in sterile disposable consumables. Bacteria collection in the intelligent fecal bacteria separation system;

(2) Fill the sampling bottle with physiological saline through the liquid filling drive device to soak the stool sample;

(3) Turn on the air pump 2 and collect the mixed bacteria in the sampling bottle into the buffer bottle through the filters with different filter values, and then pour the bacteria in the buffer bottle into the bacteria bottle through the bacteria liquid drive device and then centrifuge , After centrifugation, the supernatant is removed, and the precipitate is retained, which is the bacterial liquid precipitate after separation and purification;

(4) Add glycerin to each centrifuge tube, mix it with a shaker, pre-freeze it, quickly freeze-dry the pre-freeze sample after the pre-freeze is completed, and dehydrate the freeze-dried bacterial solution to obtain a freeze-dried powder .

Further preferably, a stirring rod is provided in the sampling bottle, and in step (2), the stirring rod in the sampling bottle stirs the stool sample to make it sufficiently homogeneous.

In a preferred embodiment, the volume ratio of the freeze-dried bacterial flora and the traditional Chinese medicine extraction powder is 0.5-2:1, more preferably 1:1. Specifically, it is made by mixing the two together with a micro-mixer, and then filling it with acid-resistant hypromellose capsules.

In a more preferred embodiment, the traditional Chinese medicine extract powder is obtained by pulverizing the effective ingredients extracted from the traditional Chinese medicine formula of American ginseng, Polygonatum, Shenqu, Digupi, and Zedoary. More preferably, the weight ratio of American ginseng, Polygonatum, Shenqu, Digupi, and Zedoary in the traditional Chinese medicine formula is 8-12:12-18:12-18:8-12:8-12, preferably 10: 15:15:10:10.

The extraction of the active ingredients of the medicine can be carried out according to conventional methods. In a specific embodiment, the preparation method of the Chinese medicine extraction powder is as follows:

(1) Cut or crush medicinal materials according to the proportion, and place them in a decoction container;

(2) Soak in water;

(3) Heating to boiling, leaching, separating the decoction liquid, decoction of the medicine residue 2-3 times according to the law, and collecting the decoction liquid;

(4) Alcohol extraction is performed on the decoction, preferably ethanol extraction;

(5) After drying the filtrate of alcohol extraction, it is pulverized with an ultrafine pulverizer, and it is preferably an ultrafine powder less than 10 μm.

The invention also provides the application of the traditional Chinese medicine formula bacterial group capsule in the preparation of medicines for treating type 2 diabetes.

The present invention also provides a method for preparing the above-mentioned Chinese medicine formula bacterial colony capsule, which includes the steps of pulverizing and uniformly pulverizing and mixing the freeze-dried powder after the separation of fecal bacteria and the effective components of the Chinese medicine extraction, wherein the two are mixed and the micro mixer is stirred and mixed evenly. , And further filled with acid-resistant hypromellose capsules.

The present invention further provides a traditional Chinese medicine composition for treating type 2 diabetes. The composition includes American ginseng, Polygonatum, Shenqu, Digupi, and Zedoary, wherein their weight ratio is 8-12:12-18:12 -18:8-12:8-12, preferably 10:15:15:10:10.

Regarding the therapeutic mechanism of the present invention, the inventors are based on the clinical characteristics of diabetes, which belongs to the category of "diminish thirst" in Chinese medicine. Traditional Chinese medicine believes that the main pathogenesis of Diabetes is "Yin deficiency is the main factor, and dryness and heat are the standard." Traditional Chinese medicine believes that the "heat" of Diabetes has its origins as "dry heat", "damp heat", and "stasis-heat". Among the three heats, stasis-heat is the most difficult to eliminate, and the most syndromes are changed. The main pathological factors of long-term thirst and a variety of stubborn diseases. Qi can produce body fluid, nourishing yin and moisturizing dryness while giving qi tonics, you can receive better yin-tonifying effect. Aiming at the pathogenesis of diabetes, which is based on yin deficiency and mixed with stasis and heat, the inventors have formulated a treatment method for replenishing qi and nourishing yin, clearing heat and promoting blood circulation. The extracted active ingredients of traditional Chinese medicine were crushed and delicately combined with fecal bacteria freeze-dried powder to form a capsule, and its curative effect on type 2 diabetes was explored. In the present invention, American ginseng is a monarch medicine, which nourishes qi and promotes body fluids, nourishes yin, moisturizes dryness and clears away heat, and is the basis for treating diabetes with yin deficiency. Polygonatum, Radix Digupi, and Curcuma turmeric are the ministerial medicines, among them, Polygonatum vulgaris, nourishes qi and nourishes yin, nourishes the lungs and kidneys, helps American ginseng, nourishes qi and produces fluids; Radix Digupi clears heat and cools blood, and zedoary promotes qi and breaks blood stasis, and can also be healthy. The spleen, Digu skin and turmeric are used together to clear heat and cool blood to remove blood stasis, and treat the symptoms of stasis and heat. Shenqu is a supplement, invigorating the spleen and stomach, eliminating food and accumulating accumulation, dispelling gastrointestinal heat accumulation, and promoting the smooth function of Qi. This medicine is precise and powerful, one medicine has multiple effects, the compatibility is rigorous, and it is in line with the pathogenesis. At the same time, the medicine of this prescription has been verified by modern pharmacology to regulate blood sugar, lower lipids or improve hemodynamics. Shenqu can also improve intestinal flora imbalance, and the Chinese and Western medicines are connected and convincing.

The present invention has the following beneficial effects: the present invention combines the traditional Chinese medicine extraction method and deeply integrates it with the fecal bacteria separation process to form a traditional Chinese medicine formula flora capsule, which greatly improves the therapeutic effect while changing the method of administration, and achieves high efficiency. , Comfortable treatment approach. The traditional Chinese medicine formula and the intestinal flora of the present invention are combined to prepare the Chinese medicine formula flora capsule. Clinical verification shows that its therapeutic effect is significant. The combination of the two has complementary effects, which not only makes up for the decoction and administration of Chinese medicine The inconvenience and shortcomings of the infusion also overcome the pain caused by the colonoscopy during the infusion of the flora, and the treatment effect has also been significantly improved. It not only meets people's requirements for application, safety, and effectiveness, but also realizes people's desire for environmental protection, humanity, and convenience.

The present invention will be further clarified through the detailed description of the specific embodiments below, but it is not a limitation of the present invention, and is only an example for illustration.

Example 1: Extraction of flora

The present invention uses an intelligent fecal bacteria separation system to automatically separate and extract disposable consumable components. The intelligent fecal bacteria separation system is a device for separating microorganisms in the contents of the cavity. The patent number is: ZL 201410606325.4, or a stool analysis pre-processing instrument, Model: FMT-5A-50/ FMT-6A-50. Disposable consumables include filter bottle 3, separated substance tank 4, sub-filling bottle 5, deodorizing device 8, liquid absorption and deodorizing buffer bottle 11, bacterial liquid pipeline 22 and so on.

The main steps of the bacterial colony extraction method of the present invention are as follows:

Collect 200g of fecal samples from healthy donors in a sterile original tank (sterile disposable consumables, the structure is shown in Figure 2), seal and send to the aseptic operation room, with filter bottle 3, separation tank 4, sub-packing bottle 5. The deodorizing device 8, the liquid absorption and deodorizing buffer bottle 11, and the bacterial liquid pipeline 22 are placed together in the intelligent fecal bacteria separation system for bacterial colony collection.

The fecal sample is soaked by adding 1000 ml of physiological saline into the sampling bottle by the liquid adding driving device 7, and the fecal sample is stirred by the stirring device to make it sufficiently homogeneous.

Turn on the air pump 2 31 and pass the mixed bacterial liquid in the sampling bottle through A, B, C, D, E, F, J, H, I, J, K (see Figure 4 for specific values) filter bottles 3 with different filtration values Collected into the bottling bottle 5. Centrifuge at 5°C, remove the supernatant after centrifugation, and retain the precipitate. This precipitate is the separated and purified bacterial liquid precipitate, with a total weight of about 100g.

Add about 10ml (12.6g) of glycerin to each sub-bottle 5, mix it with a shaker, and place it in the refrigerator at -80°C for one hour. After the pre-freezing is completed, quickly transfer the pre-frozen sample into the bacterial liquid preparation freeze dryer (for example, product name: freeze dryer, manufacturer: Ningbo Xinzhi Biotechnology Co., Ltd., model: scientz-18N) for freeze drying at a pressure of 0.006Pa, the time is not less than 10 hours, the freeze-dried bacterial liquid is dehydrated to obtain the bacterial flora freeze-dried powder.

Example 2: Chinese medicine extraction

The effective ingredients of traditional Chinese medicine are extracted by traditional concentrated extraction method (please refer to Figure 3 for the extraction process of traditional Chinese medicine). The specific method is as follows:

According to the weight ratio of 10:15:15:10:10, take the medicinal materials American ginseng, Polygonatum, Shenqu, Digu skin, and turmeric, appropriately chop or crush, and place in a decoction container.

Add appropriate amount of water to immerse the medicinal materials for a suitable time.

Heat to boiling, soak for a certain period of time, separate the decoction liquid, decoct the medicine residue 2-3 times according to the law, and collect the decoction liquid.

Evaporate the water decocting liquid to a specific gravity of 1:1, add 2 times ethanol after cooling, mix well, let it stand overnight to precipitate, filter the clear liquid the next day, wash the precipitate with a small amount of 60% ethanol, and wash the liquid Combine the filtrate with the filtrate and recover the ethanol under reduced pressure. When it is concentrated to a concentration of 1g/ml, place it in a cold place (or add a certain amount of water and mix well) and let it stand for a certain period of time to complete the precipitation and filter it.

The filtrate is dried with a vacuum dryer, and then pulverized with an ultrafine pulverizer after drying to obtain 10μm ultrafine powder.

Example 3: Preparation of Capsules of Chinese Medicine Formula Bacteria

The process flow of the Chinese medicine formula flora is shown in Figure 5. The lyophilized powder of the flora in Example 1 is mixed with the extracted powder of Chinese medicine in Example 2 in a ratio of 1:1 by volume, and then stirred and mixed with a micro mixer. The Chinese medicine formula flora was prepared and filled with acid-resistant hypromellose capsules, each capsule containing 1.5 grams of Chinese medicine powder and 1.5 grams of bacterial powder, and then packaged in a sealed packaging bag and stored in a refrigerator at -20°C.

Example 4: Therapeutic effect of Chinese medicine formula Bacteria capsule on type 2 diabetes model rats

Through experimental research, the effect of the Chinese medicine formula bacteria capsule of the present invention on the glucose and lipid metabolism and insulin resistance of type 2 diabetes model rats is explored.

1 Experimental materials

1.1 Experimental animals

40 SPF male SD rats, weighing 200±20 grams, and 8 weeks old. Each cage is housed in separate cages for 5 animals. Humidity is 40%-60%, room temperature is 16-26℃. They are fed with ordinary feed, free to eat and water, and fed before dark. The food intake and water intake of the rats were dynamically observed, and the rats were weighed once a week. Replace the cage bedding once a day.

1.2 Drugs and reagents

The traditional Chinese medicine formula bacterial colony capsule prepared in Example 3 has a concentration of 2.1 g/ml when used, which is the maximum concentration of the medicinal solution that can pass through the rat gastric applicator.

Pioglitazone (Ruitong, 15mg/tablet, Jiangsu Hengrui Pharmaceutical Co., Ltd., National Medicine Standard: H20040631).

Streptozotocin (STZ) (product of sigma company in the United States). Glucose determination kit (Shanghai Rongsheng Biopharmaceutical Co., Ltd.), insulin ELISA kit (produced by British R&G company, Shanghai Yuanxiang Medical Equipment Co., Ltd.). Trisodium citrate, citrate, sodium pentobarbital.

1.3 High-sugar and high-fat feed formula

Cholesterol, lard, sucrose, egg yolk powder, ordinary feed.

1.4 Main instruments and equipment

Blood glucose meter and blood glucose test strips (Johnson & Johnson Medical Equipment Co.), automatic immune turbidity analyzer. Benchtop centrifuge, several syringes, test tubes, beakers and measuring cylinders, precision electronic balances, electronic scales, tin foil paper, etc.

2 Experimental method

2.1 Model making

After 1 week of adaptive feeding, they were randomly divided into a normal control group (10) and a model group (30). The normal control group was fed with ordinary feed, and the model group was fed with high-sugar and high-fat feed for 6 weeks. Rats were injected with a single injection of streptozotocin (STZ, 30mg/kg) into the abdominal cavity, and the normal control group (group A) was injected with the same amount of citrate buffer solution. The blood was collected 72 hours later and measured with the Johnson & Johnson SureStepPlus blood glucose meter. Whole blood glucose, blood glucose ≥16.7mmol/L, is determined to be a rat model of type 2 diabetes.

2.2 Animal grouping and administration

After forming the model, the model rats were randomly divided into model control group (group B), pioglitazone group (group C), and Chinese medicine formula bacteria group (group D), each with 10 rats. There were 10 normal control groups (group A). Group C was given pioglitazone 2.5g/kg•d by gavage, group D was given traditional Chinese medicine formula bacterial group capsule solution 12g/kg•d by gavage; A and B groups were given the same amount of normal saline by gavage. The administration volume is 10ml/kg body weight, once a day, for a fixed time, for 8 weeks of continuous administration.

During the experiment, eating and drinking were not restricted, and insulin and other hypoglycemic drugs were not used.

2.3 Blood specimen collection

The rats were sacrificed before model building and the next day after 8 weeks of intragastric administration. The rats were fasted for 12 hours before sacrifice to determine body weight, body length and tail length; blood samples were collected from the orbit for testing of biochemical indicators such as insulin and blood sugar.

2.4 Detection of biochemical and immune indicators

The blood glucose is detected by an automatic biochemical analyzer. All measurement items are strictly operated in accordance with the kit instructions.

2.5 Statistical processing

Observe and compare the changes of each observation index in different periods of each group, and use SPSS for Windows 22.0 statistical software for hypothesis testing and correlation analysis. All measurement data are expressed as mean±standard deviation (±SD), the comparison between the two groups and before and after using the t test, and the comparison between multiple groups using the analysis of variance (Oneway-ANOVA). P<0.05 indicates that the difference is statistically significant.

Experimental result

1 The effect of Chinese medicine formula Bacteria capsule preparation solution on fasting blood glucose of model rats

±s) 項目 FBG（mmol/l） 治療前 治療後 正常組(n=10) 5.52±0.79 5.55±0.50 模型組(n=10) 18.29±1.85 19.44±2.19 吡格列酮組(n=10) 19.25±1.59 10.05±1.19★⊿ 中藥配方菌群組(n=10) 19.83±1.43 9.42±1.09★⊿ 注：治療前後組內比較★P＜0.01；與模型組比較⊿P＜0.01。After treatment, the fasting blood glucose of the traditional Chinese medicine formula bacteria group and the pioglitazone group were significantly different from those before treatment (p<0.01). After treatment, the fasting blood glucose of the traditional Chinese medicine formula bacteria group, pioglitazone group and the model group were also significantly different. Sexual difference (p<0.01). The results are shown in Table 1 and Figure 6. Table 1: The effect of traditional Chinese medicine formula Bacteria capsule preparation solution on fasting blood glucose in type 2 diabetes model rats (

±s) project FBG (mmol/l) before therapy After treatment Normal group (n=10) 5.52±0.79 5.55±0.50 Model group (n=10) 18.29±1.85 19.44±2.19 Pioglitazone group (n=10) 19.25±1.59 10.05±1.19★⊿ Chinese medicine formula bacteria group (n=10) 19.83±1.43 9.42±1.09★⊿ Note: Comparison within the group before and after treatment ★P＜0.01; comparison with the model group ⊿P＜0.01.

2 The effect of Chinese medicine formula Bacteria capsule preparation liquid on serum fasting insulin in model rats

±s) 項目 INS(U/ml) 治療前 治療後 正常組(n=10) 19.71±1.34 20.30±0.94 模型組(n=10) 11.05±2.30 11.25±2.05 吡格列酮組(n=10) 11.17±1.89 17.94±2.57★⊿ 中藥配方菌群組(n=10) 10.89±1.77 18.73±2.66★⊿ 注：治療前後組內比較★P＜0.01；與模型組比較⊿P＜0.01。After treatment, the serum fasting insulin level of rats in the Chinese medicine formula bacteria group and the pioglitazone group increased significantly compared with that before the treatment, which was statistically significant (P＜0.01). There was also a significant increase between the two groups and the type 2 diabetes model group. Sexual difference (P<0.01). The results are shown in Table 2 and Figure 7. Table 2: Effect of Chinese medicine formula Bacteria capsule preparation solution on serum fasting insulin in type 2 diabetes model rats (

±s) project INS(U/ml) before therapy After treatment Normal group (n=10) 19.71±1.34 20.30±0.94 Model group (n=10) 11.05±2.30 11.25±2.05 Pioglitazone group (n=10) 11.17±1.89 17.94±2.57★⊿ Chinese medicine formula bacteria group (n=10) 10.89±1.77 18.73±2.66★⊿ Note: Comparison within the group before and after treatment ★P＜0.01; comparison with the model group ⊿P＜0.01.

Analysis of experimental results: Animal experiments show that there is a significant difference in fasting blood glucose between the group of Chinese medicine formula bacteria after treatment and that before treatment (p<0.01), and there is also a significant difference compared with the model group (p<0.01). It is suggested that the Chinese medicine formula bacteria capsule has the effect of lowering blood sugar.

The fasting serum insulin level of rats in the Chinese medicine formula group after treatment increased compared with that before treatment, which was statistically significant (P<0.01), and there was also a significant difference compared with the model group (P<0.01). It is suggested that the Chinese medicine formula Bacteria capsule has the effect of promoting insulin secretion and increasing fasting insulin level, which may achieve the result of lowering blood sugar through this effect.

Example 5: Observation of clinical curative effect of Chinese medicine formula Bacteria capsules on type 2 diabetes

In order to verify the effect of the Chinese medicine formula Bacteria capsules in the treatment of type 2 diabetes, we selected 60 patients to perform the Chinese medicine formula Bacteria capsules (the traditional Chinese medicine formula Bacteria capsules prepared in Example 3) (Clinical treatment of type 2 diabetes) Observation, all patients were randomly divided into two groups, including 30 cases in the treatment group and 30 cases in the control group. Before treatment, there was no significant difference between the two groups in terms of gender, age, course of disease, body mass index, etc., and they were comparable. sex.

1. Treatment plan

Both groups of patients received routine basic treatment, including health education, diet control, and regular exercise.

Control group: basic treatment + conventional western medicine (according to "Guidelines for Prevention and Treatment of Type 2 Diabetes in China (2010 Edition)") treatment.

Treatment group: basic treatment + conventional western medicine (according to "Guidelines for the Prevention and Treatment of Type 2 Diabetes in China (2010 Edition)") treatment + Chinese medicine formula bacteria capsules, 2 times a day, 20 capsules each time.

Both groups took 4 weeks as a course of treatment, and observed 3 courses of treatment.

2. Western medicine diagnostic criteria:

(1) Diabetes [According to the Diabetes Diagnosis Standards established by the World Health Organization (WHO) in 1999].

① There are obvious symptoms of DM "three more and one less", random blood glucose ≥ 11.1mmol/L (200mg/dl);

②Fasting intravenous plasma glucose ≥7.0mmol/L (126mg/dl);

③OGTT (oral 75g glucose) 2 hours PG≥11.1mmol/L (200mg/dl)

The above three items, if they meet one of them individually, can be used as the basis and criteria for diagnosis, but the diagnosis can be established when the results of the reexamination at intervals are required to still meet the diagnostic criteria.

(2) Diagnostic criteria for type 2 diabetes

According to the classification criteria of the etiology of diabetes published in Diabetes Care in July 1997, those who meet the diagnostic criteria for diabetes and the results of the insulin release test meet one of the following conditions are diagnosed as type 2 diabetes.

①Insulin resistance is predominant with relative insulin deficiency.

② Obviously lack of insulin with insulin resistance.

3. Inclusion criteria

(1) Between 18 and 65 years old;

(2) Informed consent;

(3) Meet the diagnostic criteria for type 2 diabetes;

(4) There is no restriction on gender.

4. Exclusion criteria

(1) Elevated blood sugar caused by T1DM, pregnancy, drugs and other factors;

(2) Those who fail to take the medicine according to the regulations, cannot judge the curative effect, or have incomplete data that affect the judgment of curative effect;

(3) Patients who are participating in clinical studies of other drugs.

5. Changes of blood glucose and glycosylated hemoglobin in the two groups before and after treatment

±s) 項目 對照組（n=30）   治療組（n=30） 治療前 治療後   治療前 治療後 FBG（mmol/L） 9.62±1.49 7.58±1.11★★   9.95±1.55 7.00±1.23★★⊿⊿ PBG（mmol/L） 11.82±2.77 9.43±1.70★★   12.10±2.97 8.56±1.38★★⊿ HbA1c（%） 8.38±1.39 7.49±0.91★★   8.54±1.22 7.04±0.58★⊿ 注：治療前後組內比較★P＜0.05,★★P＜0.01；組間比較⊿P＜0.05,⊿⊿P＜0.01Before treatment, there was no significant difference in blood glucose and glycosylated hemoglobin between the two groups (P>0.05), and they were comparable. After treatment, the fasting blood glucose and postprandial blood glucose of the treatment group and the control group were significantly reduced, which were significantly different from those before treatment (P<0.01); the improvement of fasting blood glucose after treatment in the treatment group was significantly different than that of the control group (P<0.01) ), the improvement of postprandial blood glucose in the treatment group was different than that of the control group (P<0.05), and the improvement of glycosylated hemoglobin in the treatment group was different than that of the control group (P<0.05). See Table 3, Figure 8, Figure 9, Figure 10. Table 3: Comparison of FBG, PBG, HbA1c before and after treatment in the two groups (

±s) project Control group (n=30) Treatment group (n=30) before therapy After treatment before therapy After treatment FBG (mmol/L) 9.62±1.49 7.58±1.11★★ 9.95±1.55 7.00±1.23★★⊿⊿ PBG (mmol/L) 11.82±2.77 9.43±1.70★★ 12.10±2.97 8.56±1.38★★⊿ HbA1c (%) 8.38±1.39 7.49±0.91★★ 8.54±1.22 7.04±0.58★⊿ Note: Comparison within groups before and after treatment ★P＜0.05, ★★P＜0.01; comparison between groups ⊿P＜0.05,⊿⊿P＜0.01

6. Result analysis

After treatment, the fasting blood glucose, postprandial blood glucose, and glycosylated hemoglobin of the treatment group and the control group were significantly reduced, which were different from those in the control group (P＜0.05 or P＜0.01), indicating that conventional western medicine + traditional Chinese medicine formula bacteria capsules can affect the blood sugar of type 2 diabetes The effect of improving blood sugar is better than that of using conventional western medicine alone to control blood sugar.

Example 6: Clinical case

The typical cases in the clinical efficacy observation according to Example 5 are provided as follows. The treatment plan of the case is: take 20 capsules each time, 2 times a day; every 4 weeks is a course of treatment; treatment is three courses of treatment.

Case 1:

Patient Zhang, male, 72 years old, has a history of "type 2 diabetes" for more than 30 years. He has taken oral "Gliclazide 30mg qd, Acarbose 50mg tid" to control blood sugar, but the blood sugar control is not good. 5mg qd”, fasting blood glucose is controlled at 8.5-9.5mmol/l, postprandial blood glucose is 10-12mmol/l, and glycosylated hemoglobin is 7-7.5%. Dry mouth and more drinking are obvious, fatter, dry stool, fatigue easily, bitter mouth, red tongue, yellow coating, thin pulse. Taking the Chinese medicine formula bacterial capsule of the present invention for three courses of treatment, dry mouth, bitter mouth, fatigue and fatigue are obviously improved, and stool is once a day. The fasting blood glucose is controlled at around 7mmol/l, the postprandial blood glucose is around 8mmol/l, and the glycosylated hemoglobin is 6.8%.

Case 2:

The patient Wang Moumou, female, 69 years old, with a history of "type 2 diabetes" for 18 years, had oral administration of "metformin 500mg tid" to control blood sugar. Later, due to obvious gastrointestinal symptoms, she switched to "saxagliptin 5mg qd, repaglinide" 1mg tid", fasting blood glucose is controlled at 9-10mmol/l, postprandial blood glucose is 9-12mmol/l, glycosylated hemoglobin is about 8.5%, blood glucose fluctuates greatly, hypoglycemia has occurred, and then I dare not control diet excessively. Eating too much is easy to be hungry, abdominal distension, loose stools, loose stools, fatigue, dry mouth, red tongue, less coating, thin pulse. Taking the Chinese medicine formula bacterial capsule of the present invention for three courses of treatment, the hunger is obviously reduced, the abdominal distension and stool conditions are obviously improved, the fasting blood sugar is controlled at 6-9mmol/l, the postprandial blood sugar is about 10mmol/l, and the glycosylated hemoglobin is 7.8%.

Case 3:

Patient Lu Mou, male, 56 years old, with a history of "type 2 diabetes" for 10 years, controlled blood glucose by oral administration of "metformin and saxagliptin", fasting blood glucose was controlled at 8-10mmol/l, postprandial blood glucose was 9-11mmol/l, glycated Hemoglobin is 7.7%, dry mouth and more drinking, bloating and loose stools. After taking the Chinese medicine formula bacteria capsule of the present invention for three courses of treatment, the conditions of dry mouth, abdominal distension and stool were significantly improved, and metformin was stopped, fasting blood glucose was controlled at 6-7mmol/l, postprandial blood glucose was about 9mmol/l, and glycosylated hemoglobin was 7.1 %.

Case 4:

Patient Jin Moumou, male, 81 years old, has a history of "type 2 diabetes" for more than 30 years. He has taken a variety of hypoglycemic drugs to control blood sugar, but the control is not good. 5 years ago, he switched to "Novoline 30R early 26u late 12u" joint" Sitagliptin 100mg qd" controls blood sugar, fasting blood sugar is controlled at 7-10mmol/l, postprandial blood sugar is 8-14mmol/l, glycosylated hemoglobin is about 7.5%, blood sugar fluctuates, poor diet control, easy to anxiety, less exercise, and abdominal distension , Stool one line for 2-3 days, red tongue, yellow coating, stringy pulse. Taking the Chinese medicine formula bacterial capsule of the present invention for three courses of treatment, the symptoms are significantly improved, the fasting blood sugar is controlled at 6-9 mmol/l, the postprandial blood sugar is about 9-10 mmol/l, and the glycosylated hemoglobin is 6.4%. The daily dosage of insulin was reduced by 4u.

Case 5:

Patient Li Moumou, male, 43 years old, with a history of "type 2 diabetes" for 3 years, oral "metformin 1000mg/day" to control blood sugar, fasting blood sugar is good, glycosylated hemoglobin 6.5%, but the gastrointestinal symptoms are obvious, abdominal distension, easy loose stools, The tongue is pale red with tooth marks, and the fur is white and greasy. Because the blood sugar control is relatively satisfactory, I am not willing to change the hypoglycemic agent temporarily. I took the Chinese medicine formula microbial capsule of the present invention for three courses of treatment. Abdominal distension and loose stools were improved. Metformin was discontinued and the glycosylated hemoglobin was 6.0%.

Case 6:

Patient Lin Mou, male, 76 years old, with a history of "type 2 diabetes" for 20 years, usually uses "insulin glargine subcutaneous injection 18u/day" + "acarbose oral 150mg/day" to control blood sugar, fasting blood sugar is controlled at 6 8mmol/l, blood glucose after meal 8-12mmol/l, blood glucose 2 hours after breakfast is about 11mmol/l, blood glucose control is acceptable during the rest of the period, glycosylated hemoglobin 7.2%, dry mouth and polydipsia, dry stools, sticky toilet, red tongue Dry quality, less moss, thin pulse. After taking the traditional Chinese medicine formula bacterial capsule of the present invention, the condition of dry mouth and stool was significantly improved after three courses of treatment, the self-reported body was relaxed, the blood sugar after breakfast was 7-10mmol/l, and the glycosylated hemoglobin was 7%.

Case 7:

Patient Li, male, 51 years old, diagnosed with "Type 2 Diabetes" for 1 month. During physical examination, it was found that the blood sugar was elevated, and the fasting reached 12mmol/l. He went to the hospital for examination and diagnosed "Type 2 Diabetes" with 9.8% glycosylated hemoglobin. The thirst was obvious. At that time, "Novoray 30 morning 22u night 10u subcutaneous injection" combined with "Saxagliptin 5mg/day" was given to control blood sugar, and with the traditional Chinese medicine formula bacterial capsule of the present invention, the treatment was three courses, and the blood sugar control was better. After three courses of treatment, thirst was significantly improved, and glycosylated hemoglobin was 6.4%.

Case 8:

Patient Zhang, female, 70 years old, diagnosed with "Type 2 diabetes" for 8 years, usually uses "Repaglinide 1mg tid" + "Saxagliptin 5mg qd" to control blood sugar, fasting blood sugar is controlled at 7-8mmol/l, Postprandial blood glucose 8-11mmol/l, glycosylated hemoglobin 7.3%, dry mouth and polydipsia, dry and loose stools, red tongue with little coating, narrow pulse. Cooperating with the traditional Chinese medicine formula bacteria capsule of the present invention, the dry mouth and stool conditions were significantly improved for three courses of treatment, fasting blood glucose was 6-8 mmol/l, postprandial blood glucose was 7-9 mmol/l, and glycosylated hemoglobin was 6.4%.

Case 9:

Patient Lu Moumou, female, 81 years old, diagnosed with "type 2 diabetes" for nearly 40 years, usually uses "Novoray 30 early 26u late 14u" subcutaneous injection to control blood sugar, but the diet is irregular and he prefers snacks, so blood sugar control is not Good, large fluctuations, between 7-16mmol/l, difficult diet control, glycated hemoglobin 8.4%, "diabetic retinopathy, peripheral neuropathy" symptom, dry mouth and polydipsia, easy abdominal distension, numbness of hands and feet, poor vision Clear, itchy skin, decreased memory, easy mouth ulcers, constipation, red tongue, thin yellow coating, stringy pulse. Instruct the standardized use of insulin, adjust the injection dose when eating more or when the blood sugar is high, and take the traditional Chinese medicine formula microflora capsule of the present invention for three courses of treatment, dry mouth, blurred vision, improved skin itching, unobstructed stool, only eating habits It is difficult to change, blood sugar 7-11mmol/l, glycosylated hemoglobin 7%.

Case 10:

Patient Nan, male, 50 years old, diagnosed with "Type 2 diabetes" for 12 years, usually uses "Insulin glargine 20u/day subcutaneous injection" + "Novolong 1mg oral before meals" to control blood sugar, has "indigestion", easy Abdominal distension. Two years ago, the possibility of "gastroparesis" was considered. The blood sugar fluctuates between 6-13mmol/l. If you eat more indigestible food, you will have abdominal distension, even vomiting, irregular bowel movements, one line for 1-4 days, sometimes dry Time is thin, the tongue is pale red with tooth marks, the fur is white and greasy, and the pulse is weak. Instructed regular diet, combined with the traditional Chinese medicine formula microbial capsule of the present invention, treated for three courses, the abdominal distension was significantly improved, no vomiting, the stool was significantly improved, the blood sugar was 6-11mmol/l, and the glycosylated hemoglobin was 7.5%.

Case 11:

Patient Chen Moumou, female, 67 years old, diagnosed with "type 2 diabetes" for nearly 20 years, had been treated with oral hypoglycemic drugs, and later due to poor blood sugar control, now using "Insulin Glargine 14u/day subcutaneous injection" + "Aka Chew and take 50mg before three meals of wave sugar to control blood sugar. Because of daily insulin injection, the mood is anxious, fasting blood sugar is about 7mmol/l, postprandial blood sugar is 9-10mmol/l, glycosylated hemoglobin is 8%, and the Chinese medicine formula of the present invention is taken in conjunction with the flora Capsules, three courses of treatment, mood anxiety improved, insulin glargine dosage reduced to 10u/day, fasting blood glucose around 7mmol/l, postprandial blood glucose 7-9mmol/l, glycosylated hemoglobin 7.5%.

Case 12:

Patient Zhang Moumou, male, 37 years old, physical examination found elevated blood sugar, later diagnosed as "type 2 diabetes", found fasting blood sugar 7.2mmol/l, postprandial blood sugar 10.0mmol/l, glycosylated hemoglobin 7.8%, polyphagia easy to hunger , Dry mouth, guide diet control, proper exercise, take the Chinese medicine formula microbial capsule of the present invention, treat for three courses, eat more and be hungry, improve dry mouth, fasting blood sugar about 6.5mmol/l, postprandial blood sugar about 8.0mmol/l , Glycated hemoglobin 7.3%.

without

Figure 1 is a schematic diagram of the structure of the intelligent fecal bacteria separation system used in the present invention (ie Figure 1 in the 201410606325.4 patent).

Figure 2 is a schematic diagram of the structure of the sterile disposable consumable of the present invention.

Figure 3 is a schematic flow diagram of the extraction process of traditional Chinese medicine.

Figure 4 is a schematic diagram of the structure of the stainless steel filter in the intelligent fecal bacteria separation system of the present invention. Among them, Class A is 1.0mm, Class B 600µm, Class C 500µm, Class D 350µm, Class E 250µm, Class F 200µm, Class G 150µm, Class H 100µm, Class I 70µm, Class J 60µm, Class K 40µm.

Fig. 5 is a schematic diagram of the process flow of the bacterial flora of the Chinese medicine formula of the present invention.

Figure 6 shows the effect of the formulation of the Chinese medicine formula Bacteria Capsules on the fasting blood glucose of type 2 diabetes model rats.

Figure 7 shows the effect of the Chinese medicine formula Bacteria capsule preparation solution on the serum insulin of type 2 diabetes model rats.

Figure 8 shows the comparison of FBG between the two groups before and after treatment.

Figure 9 shows the comparison of PBG between the two groups before and after treatment.

Figure 10 shows the comparison of HbA1c between the two groups before and after treatment.

Claims (10)

The Chinese medicine formula bacterial colony capsule is prepared by pulverizing the freeze-dried powder after the separation of fecal bacteria and the Chinese medicine extraction active ingredient to obtain the Chinese medicine extraction powder. The traditional Chinese medicine formula bacterial colony capsule according to claim 1, wherein the method for separating fecal bacteria is to separate the contents of the cavity by a microorganism separating device. The traditional Chinese medicine formula flora capsule according to claim 2, wherein the preparation method of the freeze-dried powder after fecal bacteria separation includes the following steps: (1) Use a sterile sampling bottle to collect stool samples from healthy donors, seal them and send them to the sterile operating room, and place them in series with the filter bottles, AB bottles, deodorizing bottles, buffer bottles, and bacteria liquid bottles in sterile disposable consumables. Bacteria collection in the intelligent fecal bacteria separation system; (2) Fill the sampling bottle with physiological saline through the automatic liquid filling device of the liquid filling drive device to soak the stool sample; (3) Turn on the second air pump and collect the mixed bacteria in the sampling bottle into the buffer bottle through filters with different filtration values, and then pour the bacteria in the buffer bottle into the bacteria bottle through the bacteria liquid drive device and then centrifuge After centrifugation, the supernatant is removed and the precipitate is retained, which is the bacterial liquid precipitate after separation and purification; (4) Add glycerin to each centrifuge tube, mix it with a shaker, pre-freeze it, quickly freeze-dry the pre-freeze sample after the pre-freeze is completed, and dehydrate the freeze-dried bacterial solution to obtain a freeze-dried powder . The traditional Chinese medicine formula bacterial colony capsule according to claim 3, wherein the sampling bottle is provided with a stirring rod, and in step (2), the stirring rod in the sampling bottle stirs the stool sample to make it fully homogeneous. The traditional Chinese medicine formula bacterial colony capsule according to any one of claims 1 to 4, wherein the volume ratio of the lyophilized bacterial colony powder and the traditional Chinese medicine extract powder is 0.5-2:1, more preferably 1:1; wherein the bacteria The mass freeze-dried powder and the traditional Chinese medicine extraction powder are mixed uniformly by mixing the two together and then mixing them with a micro mixer. According to claim 5, the Chinese medicine formula bacterial colony capsule is further filled with acid-resistant hypromellose capsules to obtain the Chinese medicine formula bacterial colony capsule. The Chinese medicine formula flora capsule according to any one of claims 1 to 4, wherein the Chinese medicine extract powder is obtained by pulverizing active ingredients extracted from the Chinese medicine formula of American ginseng, Polygonatum, Shenqu, Digupi, and Zedoary. The Chinese medicine formula bacterial group capsule according to claim 7, wherein the weight ratio of American ginseng, Polygonatum, Shenqu, Digupi, and Zedoary in the Chinese medicine formula is 8-12:12-18:12-18:8-12: 8-12, preferably 10:15:15:10:10. The Chinese medicine formula bacterial colony capsule according to claim 7, wherein the preparation method of the Chinese medicine extract powder is as follows: (1) Cut or crush medicinal materials according to the proportion, and place them in a decoction container; (2) Soak in water; (3) Heating to boiling, leaching, separating the decoction liquid, decoction of the medicine residue 2-3 times according to the law, and collecting the decoction liquid; (4) Alcohol extraction is performed on the decoction, preferably ethanol extraction; (5) After drying the filtrate of alcohol extraction, it is pulverized with an ultrafine pulverizer, and it is preferably an ultrafine powder less than 10 μm. Application of the Chinese medicine formula bacterial group capsule according to any one of claims 1 to 9 in the preparation of a medicine for treating type 2 diabetes.

Images