Hypoglycemic traditional Chinese medicine composition for inhibiting metabolic inflammation and preparation method of hypoglycemic traditional Chinese medicine composition

Abstract

The invention discloses a blood-glucose-reducing traditional Chinese medicine composition for inhibiting metabolic inflammation and a preparation method of the blood-glucose-reducing traditional Chinese medicine composition. The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 20-40 parts of herba eupatorii, 5-7 parts of rhizoma coptidis, 10-20 parts of ginseng, 10-20 parts of poria cocos, 10-20 parts of roasted rhizoma atractylodis macrocephalae, 5-7 parts of honey-fried licorice root, 10-20 parts of agastache rugosus, 8-10 parts of radix aucklandiae, 12-22 parts of radix puerariae, 8-10 parts of stiff silkworm, 10-20 parts of ramulus euonymi, 10-20 parts of ramulus mori and 2-5 parts of cortex cinnamomi. The traditional Chinese medicine composition disclosed by the invention can be used for inhibiting the proinflammatory factor IL-6 level (Plt; 0.05), increasing the level of anti-inflammatory factor IL-22 (Plt; in small intestine tissues of rats with type 2 diabetes, the traditional Chinese medicine composition disclosed by the invention can be used for relieving inflammation infiltration; immunohistochemical detection results show that the traditional Chinese medicine composition can improve the expression level of AHR and IL-22 in small intestine tissues, provides a scientific basis for inhibiting metabolic inflammation by using a detoxification and turbidity dissolving method so as to improve insulin resistance and treat type 2 diabetes, and also provides a new treatment idea and drug selection.

Description

Technical Field

[0001] The invention relates to a hypoglycemic Chinese medicinal composition for inhibiting metabolic inflammation and a preparation method thereof, belonging to the technical field of traditional Chinese medicine. Background Art

[0002] Metabolic inflammation is closely related to insulin resistance in type 2 diabetes mellitus (T2DM). It is a key component of T2DM and can lead to insulin resistance. For example, interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) can inhibit the insulin signaling pathway, leading to insulin resistance and thus inducing T2DM. Anti-inflammatory factors such as interleukin-22 (IL-22) can improve insulin sensitivity. Further experimental studies have shown that IL-22 treatment in T2DM mice infected with Mycobacterium tuberculosis can reduce serum insulin levels. Interleukin-17 (IL-17) and IL-22 produced by group 3 innate lymphoid cells (ILC3s) and Th17 cells interact with each other to regulate epithelial cell antimicrobial peptide secretion and the production of immunoglobulin A in the intestine, limiting microbial translocation and preventing systemic inflammation, thereby improving insulin resistance. Therefore, inhibiting metabolic inflammation is an important research direction in the field of improving insulin resistance in type 2 diabetes. Summary of the invention

[0003] The technical problem to be solved by the present invention is to provide a hypoglycemic Chinese medicine composition for inhibiting metabolic inflammation and a preparation method, so as to provide a new drug option for diabetic patients.

[0004] In order to solve the above technical problems, the technical solution adopted by the present invention is as follows:

[0005] A hypoglycemic Chinese medicine composition for inhibiting metabolic inflammation. The Chinese medicine composition is prepared from the following raw materials in parts by weight: 20-40 parts of perilla, 5-7 parts of coptis root, 10-20 parts of ginseng, 10-20 parts of tuckahoe, 10-20 parts of stir-fried atractylodes, 5-7 parts of roasted liquorice, 10-20 parts of patchouli, 8-10 parts of costusroot, 12-22 parts of kudzu root, 8-10 parts of bombyx batryticatus, 10-20 parts of euphorbia pulex, 10-20 parts of mulberry twig, and 2-5 parts of cinnamon bark.

[0006] As a preferred embodiment, the optimal weight of the raw materials is: 30g of Perilla frutescens, 6g of Coptis chinensis, 15g of Ginseng, 15g of Poria, 15g of Fried Atractylodes macrocephala, 6g of Rhizoma Licorice, 15g of Patchouli, 9g of Aucklandia lappa, 18g of Pueraria lobata, 9g of Bombyx batryticatus, 15g of Psoralea corylifolia, 15g of Morus alba, and 3g of Cinnamon bark.

[0007] The preparation method of the above-mentioned hypoglycemic Chinese medicine composition for inhibiting metabolic inflammation comprises the following steps: weighing the medicinal materials Perilla frutescens, Coptis chinensis, Ginseng, Poria cocos, stir-fried Atractylodes macrocephala, roasted Licorice root, Patchouli, Aucklandia lappa, Pueraria root, Bombyx batryticatus, Euphorbia obesa, Morus twig, and Cinnamon bark, soaking the above-mentioned medicinal materials in 5 times the mass of pure water for 2 hours, boiling them with high heat at 150-220°C and then decocting them with low heat at 80-120°C for 30 minutes, collecting the decoction after filtering; adding 2 times the mass of pure water to the filter residue and decocting it for 20 minutes, collecting the decoction after filtering; mixing the two decoctions and concentrating them to a Chinese medicine concentration of 2.5 g / ml to obtain the Chinese medicine composition of the present invention.

[0008] As a preferred embodiment, the above-mentioned hypoglycemic Chinese medicine composition for inhibiting metabolic inflammation can also be prepared into a pharmaceutical preparation, which is made of the following raw materials in parts by weight: 20-40 parts of Perilla frutescens, 5-7 parts of Coptis chinensis, 10-20 parts of Ginseng, 10-20 parts of Poria cocos, 10-20 parts of stir-fried Atractylodes macrocephala, 5-7 parts of roasted Licorice root, 10-20 parts of Patchouli, 8-10 parts of Costus root, 12-22 parts of Pueraria root, 8-10 parts of Bombyx batryticatus, 10-20 parts of Psoralea corylifolia, 10-20 parts of Morus twig, 2-5 parts of cinnamon, and pharmaceutically acceptable excipients.

[0009] As a preferred embodiment, the pharmaceutical preparation includes injection, oral solution, pill, powder, paste, tablet, granule, powder or capsule.

[0010] Studies have found that turbidity and toxins are related to metabolic inflammation. Turbidity and toxin precursors such as phlegm and dampness and blood stasis are closely related to metabolic inflammation. Among patients with type 2 diabetes of different syndromes, C-reactive protein and interleukin-6 are the two inflammatory factors with the highest proportion in the phlegm and dampness type and the blood stasis type. The method of resolving phlegm and removing blood stasis can significantly reduce the levels of inflammatory factors TNF-α and CRP in patients with type 2 diabetes. Some scholars even believe that the pathological substances of the metabolic inflammatory state are inflammatory factors, which can be classified as "turbidity and toxins" based on their properties. The method of detoxification and turbidity removal may improve insulin resistance in type 2 diabetes by inhibiting metabolic inflammation, and more and more studies have provided evidence. Therefore, turbidity and toxins are closely related to metabolic inflammation, and the method of detoxification and turbidity removal can improve insulin resistance in type 2 diabetes by inhibiting metabolic inflammation.

[0011] The present invention conducted experiments on inhibiting the secretion of pro-inflammatory factors and promoting the secretion of anti-inflammatory factors. The results showed that the Chinese medicine composition of the present invention can inhibit the level of pro-inflammatory factor IL-6 in the serum of type 2 diabetic rats (P<0.05), increase the level of anti-inflammatory factor IL-22 (P<0.05), and in the small intestinal tissue of type 2 diabetic rats, the Chinese medicine composition of the present invention can alleviate inflammatory infiltration; the results of immunohistochemical detection showed that the Chinese medicine composition of the present invention can increase the expression levels of AHR and IL-22 in small intestinal tissue. The present invention provides a scientific basis for using the detoxification and turbidity removal method to inhibit metabolic inflammation, thereby improving insulin resistance and treating type 2 diabetes, and also provides a new treatment idea and drug selection. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 This is a graph of protein expression levels of TLR4, NFκB, IκK, and IκB in skeletal muscle tissue of db / db mice;

[0013] Figure 2 is the pancreatic islet morphology of mice in each group;

[0014] Figure 3 This is the expression diagram of TLR4, NFκB, IκK, and IκB in pancreatic tissue;

[0015] Figure 4 is the histomorphology of small intestine of rats in each group;

[0016] Figure 5 This is the expression diagram of AHR and IL-22 in small intestinal tissue. DETAILED DESCRIPTION

[0017] 1. Experiments on inhibiting the secretion of pro-inflammatory factors

[0018] 1. Materials

[0019] 1.1 Animals

[0020] Forty SPF male 6-week-old spontaneous type 2 diabetic db / db mice and ten normal male db / m mice of the same age, weighing 28-35 g, were purchased from Changzhou Cavens Experimental Animal Co., Ltd. (certificate number: NO.201826879). This experiment was reviewed and approved by the Ethics Committee of the Institute of Experimental Animals of Guizhou University of Traditional Chinese Medicine.

[0021] 1.2 Drugs and reagents

[0022] The Chinese medicine composition (Tangduqing) of the present invention is provided by the pharmacy of the Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine, rosiglitazone (ROS) (Chengdu Hengrui Pharmaceutical Co., Ltd., approval number: H20030569), hematoxylin, histochemical primary antibody diluent, DAB color development kit, enhanced enzyme-labeled goat anti-mouse / rabbit IgG polymer, horseradish enzyme-labeled goat anti-rabbit IgG (H+L) (Beijing Zhongshan Jinqiao Biotechnology Co., Ltd., batch numbers: ZLI-9610, ZLI-9030, ZLI-9018, PV-9001, ZB-2301), Anti-TLR4, Anti-NFκB p65, Anti-IκK alpha, Anti-IκB alpha antibody (Abcam, batch numbers: ab22048, 14220-1-AP, ab32041, ab32518); Insulin kit (Nanjing Jiancheng Biological Research Institute, batch number: H203-1-2); IL-1β kit, IL-6 kit, IL-18 kit, TNF-α kit (Nanjing Jiancheng Biological Research Institute. Batch numbers: H002, H007-1-2, H015, H052-1), FINS kit, IL-22 kit (Wuhan Gene Beauty Biotechnology Co., Ltd., batch numbers: 64-17-5, JYM1204Ra), Protein Maker, reverse transcription kit, BCA protein concentration determination kit (Thermo Fisher Scientific, batch numbers: 26616, A25742, P0100), WB primary antibody diluent (Beyotime, batch number: P0023A-500ml)

[0023] 1.3 Instruments

[0024] BBS-DDC clean bench (Jinan Xinbeisi Biotechnology Co., Ltd.), centrifugal oscillator, S1010E centrifuge (Labnet, USA), low-speed centrifuge (Hunan Xiangyi Laboratory Instrument Development Co., Ltd.), fluorescence quantitative PCR instrument, electrotransfer instrument, electrophoresis tank, Bio-rad680 microplate reader (Bio-Rad, USA), 4℃, -20℃ refrigerator (Qingdao Haier Co., Ltd.), -80℃ ultra-low temperature freezer (Zhongke Meiling Cryogenic Technology Co., Ltd.), TS-8 horizontal shaker (Jiangsu Haimen Qilin Bell Instrument Manufacturing Co., Ltd.), developer (Azure, USA), slicer (Leica, Germany), TB-718E embedding machine (Hubei Taiwei Technology Industry Co., Ltd.), blood glucose meter and test strips (Roche integrated blood glucose meter, Germany).

[0025] 2. Methods

[0026] 2.1 Drug preparation

[0027] The Chinese medicine composition of the present invention is composed of 30g of Perilla frutescens, 6g of Coptis chinensis, 15g of Ginseng, 15g of Poria cocos, 15g of Rhizoma Atractylodis Macrocephalae, 6g of Radix Glycyrrhizae, 15g of Patchouli, 9g of Aucklandia lappa, 18g of Radix Puerariae, 9g of Bombyx batryticatus, 15g of Feathers of the ...

[0028] 2.2 Grouping and drug administration of experimental animals

[0029] 40 SPF male 6-week-old spontaneous type 2 diabetes db / db mice and 10 male normal db / m mice of the same age, weighing 28-35g, were randomly divided into 4 groups after adaptive feeding for 1 week, including model group (MOD), low-dose group of the Chinese medicine composition of the present invention (TDQL), high-dose group of the Chinese medicine composition of the present invention (TDQH) and rosiglitazone group (ROS), with 10 mice in each group, and normal db / m mice were set as blank control group (CON). The mice were intragastrically administered daily for 8 consecutive weeks, wherein MOD and CON were intragastrically administered with distilled water, and the dosage was 5ml / kg / d; the dosage of TDQL was 10.15g / kg.d, the dosage of TDQH was 20.30g / kg.d, and the dosage of ROS was 4mg / kg.d.

[0030] 2.3 Index detection

[0031] 2.3.1 Blood sugar

[0032] Mice were gavaged until the 8th week. All mice were fasted for 12 hours but not water. Blood was collected from the tail tip in the morning of the next day to test the fasting blood glucose of each mouse, which was recorded as the blood glucose value at 0 min. After recording, 2 g / kg / 10 ml of 50% glucose was gavaged, and blood glucose was measured at the tail tip 30 min, 60 min and 120 min after gavage.

[0033] 2.3.1TNF-α, IL-6, IL-1β, IL-18

[0034] The serum to be tested was taken out and completely thawed at room temperature. After centrifugation at 2000 r / min for 10 minutes, it was allowed to stand at room temperature for 20 minutes. According to the operating requirements of the IL-1β kit, IL-6 kit, IL-18 kit, and TNF-α kit (Nanjing Jiancheng Biological Company Research Institute), the standard was diluted, sampled, enzymes were added, incubated, washed, developed, the reaction was terminated, and the absorbance of each well (i.e., OD value) was measured.

[0035] 2.3.2 TLR4, NFκB, IκK, IκB protein and mRNA expression

[0036] Western blotting method was used to detect the expression of TLR4, NFκB, IκK, and IκB proteins in mouse skeletal muscle tissue: After taking mouse skeletal muscle tissue, protein lysis buffer was added and lysed at -80℃, thawed and incubated at 4℃ for 5min, centrifuged for 15min, and extracted and the protein concentration was determined by BCA method. 20ul protein sample was added to each well, separated by 10% SDS-PAGE electrophoresis, cut the gel, and transferred to the membrane at a constant current of 200mA for 1h. The primary antibody, namely Anti-TLR4, Anti-NFκB p65, Anti-IκKalpha, and Anti-IκB alpha antibodies (Abcam) were diluted with antibody diluent (all antibodies were used at 1:1000), incubated at 4℃ overnight, and the next day, horseradish enzyme-labeled goat anti-rabbit IgG (H+L) (Beijing Zhongshan Jinqiao Biotechnology Co., Ltd.) was diluted with antibody diluent (1:5000), incubated at 4℃ for 2h, developed with ECL chemiluminescent solution, and gray value was analyzed with Image J.

[0037] RT-PCR was used to detect the mRNA expression of TLR4, NFκB, IκK, and IκB in mouse skeletal muscle tissue: 1 g of skeletal muscle tissue was taken from each mouse, ground, lysed, and centrifuged, and then 30 ul of DEPC water was added to dissolve it, the RNA concentration was measured, and the reverse transcription reaction was performed (reverse transcription kit, Thermo Fisher Scientific). The PCR amplification system was designed with Premier 5.0 software to design gene sequence-specific primers, and the prepared amplification system was executed according to the amplification reaction conditions, and then the configured amplification product system was reacted according to the reaction conditions, and the fluorescence curve and melting curve were observed. Finally, the ct value of the target gene and the internal reference gene GAPDH of each sample was obtained according to the qPCR reaction curve, and the 2-△△ct method was used for relative quantification to calculate the relative value of the gene expression of each group.

[0038] 2.3.3 Pathological examination of pancreatic tissue in db / db mice

[0039] HE staining and immunohistochemistry were used to detect the pathological changes of pancreatic tissue in mice.

[0040] 3. Results

[0041] 3.1 Effects on blood sugar

[0042] (1) Effect on fasting (0min) blood glucose (see Table 1): Compared with the control group, the fasting blood glucose of mice in the model group, the low-dose group and the high-dose group of the Chinese medicine composition of the present invention, and the rosiglitazone group was significantly increased, and the difference was statistically significant (P<0.0001, P<0.0001, P<0.0001, P<0.0001). Compared with the model group, the low-dose group of the Chinese medicine composition of the present invention had no statistically significant difference compared with the model group (P=0.1603); the fasting blood glucose of the high-dose group of the Chinese medicine composition of the present invention and the rosiglitazone group was significantly reduced compared with the model group, and the difference was statistically significant (P=0.0259, P=0.0009). There was no significant statistical difference between the low-dose group and the high-dose group of the Chinese medicine composition of the present invention and the rosiglitazone group (P=0.2610, P=0.6832). There was no significant statistical difference between the low-dose group and the high-dose group of the Chinese medicine composition of the present invention (P=0.9356).

[0043] (2) Effect on 30-min blood glucose (see Table 1): Compared with the control group, the 30-min blood glucose of mice in the model group, the low-dose group, the high-dose group of the Chinese medicine composition of the present invention, and the rosiglitazone group was significantly increased, with significant statistical significance (P<0.0001, P<0.0001, P<0.0001, P<0.0001). Compared with the model group, the 30-min blood glucose of mice in the low-dose group, the high-dose group of the Chinese medicine composition of the present invention, and the rosiglitazone group showed a trend of decrease, and the difference was not statistically significant (P=0.6810, P=0.2074, P=0.1843). There was no significant statistical significance between the low-dose group and the rosiglitazone group (P=0.8847, P>0.9999). There was no statistical significance between the low-dose group and the high-dose group of the Chinese medicine composition of the present invention (P=0.9095).

[0044] (3) Effect on 60-min blood glucose (see Table 1): Compared with the control group, the 60-min blood glucose of mice in the model group, the low-dose group and the high-dose group of the Chinese medicine composition of the present invention, and the rosiglitazone group increased significantly, with significant statistical significance (P<0.0001, P<0.0001, P<0.0001, P<0.0001). Compared with the model group, except for the low-dose group of the Chinese medicine composition of the present invention, which had no statistically significant difference compared with the model group (P=0.0952), the high-dose group of the Chinese medicine composition of the present invention and the rosiglitazone group had a decrease in 60-min blood glucose compared with the model group, with statistically significant differences (P=0.0164, P=0.0018). There was no significant statistical significance between the low-dose group and the rosiglitazone group of the Chinese medicine composition of the present invention (P=0.5515, P=0.9087). There was no statistical significance between the low-dose group and the high-dose group of the Chinese medicine composition of the present invention (P=0.9573).

[0045] (4) Effect on 120-min blood glucose (see Table 1): Compared with the control group, the 120-min blood glucose of mice in the model group, the low-dose group, the high-dose group of the Chinese medicine composition of the present invention, and the rosiglitazone group was significantly increased, with significant statistical significance (P<0.0001, P<0.0001, P<0.0001, P<0.0001). Compared with the model group, the 120-min blood glucose of mice in the low-dose group, the high-dose group of the Chinese medicine composition of the present invention, and the rosiglitazone group was reduced in each administration group, with statistically significant differences (P=0.0137, P=0.0004, P<0.0001). There was no statistically significant difference between the low-dose group and the high-dose group of the Chinese medicine composition of the present invention and the rosiglitazone group (P=0.4700, P=0.9913). There was no statistically significant difference between the low-dose group and the high-dose group of the Chinese medicine composition of the present invention (P=0.7370).

[0046] Table 1 Comparison of blood glucose at different time points in each group of mice during oral administration of grapes

[0047]

[0048] Note: Compared with the CON group, a P<0.0001, b P<0.0001, c P<0.0001, d P<0.0001; compared with the MOD group, e P<0.05, f P<0.001, g P<0.05, h P<0.05, i P<0.05, j P<0.001, k P<0.0001.

[0049] 3.2 Effect of the Chinese medicine composition of the present invention on inflammatory factors TNF-α, IL-6, IL-1β, and IL-18 in mouse serum

[0050] In order to illustrate the effect of the Chinese medicine composition of the present invention on the inflammatory factors TNF-α, IL-6, IL-1β, and IL-18 in the serum of type 2 diabetic db / db mice, after 8 weeks of drug intervention, ELISA was used to detect TNF-α, IL-6, IL-1β, and IL-18 in each group of mice, and one-way analysis of variance was performed. The results are shown in Table 2:

[0051] (1) Effect on TNF-α: Compared with the control group, the TNF-α of mice in the model group and the low-dose group of the Chinese medicine composition of the present invention was significantly increased, and the difference was statistically significant (P<0.0001, P<0.0001), while the high-dose group of the Chinese medicine composition of the present invention and the rosiglitazone group were not statistically significant compared with the control group (P=0.0676, P=0.2108). Compared with the model group, the TNF-α of mice in the low-dose group, high-dose group and rosiglitazone group of the Chinese medicine composition of the present invention was significantly reduced, and the difference was statistically significant (P<0.0001, P<0.0001, P<0.0001). Compared with the rosiglitazone group, the TNF-α of the low-dose group of the Chinese medicine composition of the present invention was increased, which was statistically significant (P<0.0001); the high-dose group of the Chinese medicine composition of the present invention was not statistically significant compared with the rosiglitazone group (P=0.9822). Compared with the low-dose and high-dose groups of the Chinese medicine composition of the present invention, TNF-α in the high-dose group was significantly reduced, which was statistically significant (P<0.0001).

[0052] (2) Effect on IL-6: Compared with the control group, the IL-6 of mice in the model group, the low-dose group, the high-dose group and the rosiglitazone group of the Chinese medicine composition of the present invention was significantly increased, and the difference was statistically significant (P<0.0001, P<0.0001, P=0.0002, P=0.0070). Compared with the model group, the IL-6 of mice in the low-dose group, the high-dose group and the rosiglitazone group of the Chinese medicine composition of the present invention was significantly decreased, and the difference was statistically significant (P<0.0001, P<0.0001, P<0.0001). Compared with the rosiglitazone group, the IL-6 of the low-dose group of the Chinese medicine composition of the present invention was increased, which was statistically significant (P<0.0001); no significant change was observed in the high-dose group, which was not statistically significant (P=0.7668). Compared with the low-dose group and the high-dose group of the Chinese medicine composition of the present invention, the IL-6 of the high-dose group was significantly decreased, which was statistically significant (P<0.0001).

[0053] (3) Effect on IL-1β: Compared with the control group, the IL-1β of mice in the model group and the low-dose group of the Chinese medicine composition of the present invention was significantly increased, and the difference was statistically significant (P<0.0001, P=0.0023); compared with the control group, there was no significant statistical difference in the high-dose group of the Chinese medicine composition of the present invention and the rosiglitazone group (P=0.3052, P=0.5042). Compared with the model group, the IL-1β of mice in the low-dose group, the high-dose group and the rosiglitazone group of the Chinese medicine composition of the present invention was significantly reduced, and the difference was statistically significant (P<0.0001, P<0.0001, P<0.0001). Compared with the rosiglitazone group, the IL-1β of mice in the low-dose group and the high-dose group of the Chinese medicine composition of the present invention did not change much, and there was no significant statistical significance (P=0.1326, P=0.9968). There was no significant statistical difference between the low-dose group and the high-dose group of the Chinese medicine composition of the present invention (P=0.2488).

[0054] (4) Effect on IL-18: Compared with the control group, the IL-18 of the model group mice was significantly higher than that of the control group, and the difference was statistically significant (P<0.0001); the IL-18 of the mice in the low-dose and high-dose groups of the Chinese medicine composition of the present invention and the rosiglitazone group was not statistically significant compared with the control group (P=0.6568, P=0.0945, P=0.1856). Compared with the model group, the IL-18 of each treatment group was significantly reduced, and the difference was statistically significant (P<0.0001, P<0.0001, P<0.0001). There was no statistically significant difference between the low-dose and high-dose groups of the Chinese medicine composition of the present invention and the rosiglitazone group (P=0.9301, P=0.9974). There was no statistically significant difference between the low-dose and high-dose groups of the Chinese medicine composition of the present invention (P=0.7987).

[0055] Table 2 Comparison of TNF-α, IL-6, IL-1β, and IL-18 in each group of mice

[0056]

[0057] Note: Compared with the CON group, the MOD group a P<0.0001; ZDQL, ZDQH, ROS group compared with MOD group, b P<0.0001, c P<0.0001, d P<0.0001; ZDQL compared with CON, e P<0.0001, f P<0.0001, g P<0.05; ZDQH compared with CON, h P<0.05; ZDQL compared with ROS, iP<0.0001, j P<0.0001; ZDQL compared with ZDQH, k P<0.0001, l P<0.0001.

[0058] 3.3 TLR4, NFκB, IκK, and IκB protein expression

[0059] Compared with the model group, the expression of TLR4, NFκB, IκK, and IκB proteins in mice in the low-dose and high-dose groups of the Chinese medicine composition of the present invention and the rosiglitazone group was reduced (p<0.05); compared with the rosiglitazone group, the expression of IκK protein in the low-dose group of the Chinese medicine composition of the present invention was significantly increased (p<0.05), while there was no difference in the expression of TLR4, NFκB, and IκB proteins (P>0.05), and there was no significant difference in the expression of TLR4, NFκB, IκK, and IκB proteins in the high-dose group of the Chinese medicine composition of the present invention (P>0.05); compared with the low-dose and high-dose groups of the Chinese medicine composition of the present invention, the expression of IκK protein in the high-dose group was significantly reduced compared with the low-dose group (P<0.05), and the expression of TLR4, NFκB, and IκB proteins in the low-dose group showed a decreasing trend, but the difference was not statistically significant. The results are shown in Table 3. Figure 1 :

[0060] Table 3 Comparison of TLR4, NFκB, IκK, and IκB in each group of mice detected by WB

[0061]

[0062]

[0063] Note: Compared with MOD, * P<0.05; compared with ROS, # P<0.005; compared with TDQL △ P<0.05.

[0064] 3.4 Expression of TLR4, NFκB, IκK, and IκB mRNA

[0065] Compared with the model group, the expressions of TLR4, NFκB, IκK, and IκB in mice in the low-dose and high-dose groups of the Chinese medicine composition of the present invention and the rosiglitazone group were significantly reduced (P<0.05); compared with the rosiglitazone group, there were no significant differences in the expressions of TLR4, NFκB, IκK, and IκB in mice in the low-dose and high-dose groups of the Chinese medicine composition of the present invention (P>0.05); compared with the low-dose and high-dose groups of the Chinese medicine composition of the present invention, there were no significant differences in the expressions of TLR4, NFκB, IκK, and IκB (P>0.05), as shown in Table 4.

[0066] Table 4 Comparison of TLR4, NFκB, IκK, and IκB in each group of mice detected by RT-PCR

[0067]

[0068] Note: Compared with MOD, * P<0.05.

[0069] 3.5 Effect of the Chinese medicine composition of the present invention on pancreatic tissue in mice

[0070] (1) HE staining (such as Figure 2 ) as shown: the pancreatic islet structure of the normal control group mice was complete, with good morphology and clear boundaries. The islet cells were arranged neatly and regularly, the nuclei were (class) round, the size was relatively uniform, the nuclei were deeply stained, the cytoplasm was abundant, and the nuclei were centered. No inflammatory cell infiltration was observed. The pancreatic islet structure of the model group mice was incomplete, with abnormal morphology and unclear boundaries. The cells in the islets were arranged in a disordered manner, with different sizes, and were "crowded" locally. The nuclei were more (class) elliptical, with lightly stained nuclei. The cytoplasm of some cells was not abundant, and the nuclei were close to the cell membrane and vacuoles were seen. Compared with the model group, the pancreatic islet structure of the low-dose and high-dose groups of the Chinese medicine composition of the present invention and the rosiglitazone group was relatively complete, with good morphology and relatively clear boundaries. The islet cells were arranged relatively neatly and regularly, the nuclei were (class) round, the size was relatively uniform, the nuclei were deeply stained, the cytoplasm was relatively abundant, and the nuclei were centered.

[0071] (2) Immunohistochemical testing (such as Figure 3 ): The islet cytoplasm in the islet cell clusters of the normal group mice was obviously brownish yellow, and only the islet cell nuclei were partially colored. The islet cell nuclei in the islet cell clusters of the model group mice were obviously brownish yellow, and the cytoplasm was almost not colored. Compared with the model group, the islet cytoplasm in the islet cell clusters of the low-dose and high-dose groups of the Chinese medicine composition of the present invention and the rosiglitazone group was darker, and the islet cell nuclei were lighter than those in the model group.

[0072] 2. Experiments to promote the secretion of anti-inflammatory factors

[0073] 1 Experimental Materials

[0074] 1.1 Experimental instruments

[0075] BBS-DDC clean bench (Jinan Xinbeisi Biotechnology Co., Ltd.), centrifugal oscillator, S1010E centrifuge (Labnet, USA), low-speed centrifuge (Hunan Xiangyi Laboratory Instrument Development Co., Ltd.), fluorescence quantitative PCR instrument, electrotransfer instrument, electrophoresis tank, Bio-rad680 microplate reader (Bio-Rad, USA), 4℃, -20℃ refrigerator (Qingdao Haier Co., Ltd.), -80℃ ultra-low temperature freezer (Zhongke Meiling Cryogenic Technology Co., Ltd.), TS-8 horizontal shaker (Jiangsu Haimen Qilin Bell Instrument Manufacturing Co., Ltd.), developer (Azure, USA), slicer (Leica, Germany), TB-718E embedding machine (Hubei Taiwei Technology Industry Co., Ltd.), blood glucose meter and test strips (Roche integrated blood glucose meter, Germany).

[0076] 1.2 Experimental drugs and reagents

[0077] The Chinese medicine composition (Tangduqing, TDQ) of the present invention is composed of 30g of Perilla frutescens, 6g of Coptis chinensis, 15g of Ginseng, 15g of Poria cocos, 15g of stir-fried Atractylodes macrocephala, 6g of Rhizoma Licorice, 15g of Patchouli, 9g of Aucklandia lappa, 18g of Pueraria root, 9g of Bombyx batryticatus, 15g of Psoralea corylifolia, 15g of Morus alba and 3g of Cinnamomum cassia. The medicinal materials are soaked in 5 times the weight of pure water for 2 hours, boiled with high heat (150-220°C), then decocted with low heat (80-120°C) for 30min, filtered and collected the decoction; the filter residue is added with 2 times the weight of pure water and decocted for 20min, filtered and obtained the decoction; all the decoctions are mixed and concentrated to a Chinese medicine concentration of 2.5g / ml to obtain the Chinese medicine composition, which is prepared by the pharmacy of the Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine. Rosiglitazone (ROS) (Chengdu Hengrui Pharmaceutical Co., Ltd., approval number: H20030569), hematoxylin, histochemical primary antibody diluent, DAB color development kit, enhanced enzyme-labeled goat anti-mouse / rabbit IgG polymer, horseradish enzyme-labeled goat anti-rabbit IgG (H+L) (Beijing Zhongshan Jinqiao Biotechnology Co., Ltd., batch numbers: ZLI-9610, ZLI-9030, ZLI-9018, PV-9001, ZB-2301), Anti-TLR4, Anti-NFκB p65, Anti-IκK alpha, Anti-IκB alpha antibody (Abcam, batch numbers: ab22048, 14220-1-AP, ab32041, ab32518); Insulin kit (Nanjing Jiancheng Biological Research Institute, batch number: H203-1-2), Protein Maker, reverse transcription kit, BCA protein concentration determination kit (Thermo Fisher Scientific, batch numbers: 26616, A25742, P0100), WB primary antibody diluent (Beyotime, batch number: P0023A-500ml).

[0078] 2. Experimental Methods

[0079] 2.1 Construction of animal model

[0080] (1) Sixty 6-week-old SPF male SD rats weighing 180-220 g were purchased from Changsha Tianqin Biological Co., Ltd. with a certificate number of SCXK (Xiang) 2019-0014. All experimental rats were kept in the Animal Research Laboratory of Guizhou University of Traditional Chinese Medicine after passing the test. The breeding environment was: 12 h light and dark alternation per day, relative temperature of 23 °C ± 3 °C, and humidity of 55% ± 5%.

[0081] (2) After one week of adaptive feeding, all rats were weighed, numbered, and randomly divided into two groups: a normal feed group of 10 rats, which were fed a normal feed; and a high-fat feed group of 50 rats, which were fed a high-sugar and high-fat feed (consisting of 18% lard, 3% egg yolk powder, 20% white sugar, and 59% normal feed). The rats were allowed to drink water and eat freely. All feeds were purchased from Changsha Tianqin Biological Co., Ltd.

[0082] (3) After 5 weeks, the rats were fasted overnight but not watered for 12 hours and weighed the next day. The high-fat diet group was intraperitoneally injected with 30 mg / kg 1% streptozocin (STZ); the normal diet group was intraperitoneally injected with the corresponding volume of sodium citrate buffer (pH 4.5).

[0083] (4) Criteria for successful modeling: 72 hours after modeling, blood was collected from the tail tip vein to measure random blood sugar. If blood sugar was ≥16.67mmol / L for two consecutive days, it was considered a successful modeling of type 2 diabetes. 50 modeling rats were included in this experiment, 44 of which were successfully modeled. The remaining 6 rats were not included in the experiment because their blood sugar did not meet the standard. The modeling success rate of this experiment was 88%.

[0084] 2.2 Experimental Animal Grouping and Dosing

[0085] 44 successfully modeled type 2 diabetic rats were randomly divided into 4 groups, each with 11 rats, namely, model group (MOD), metformin hydrochloride group (MH), low-dose group of the Chinese medicine composition of the present invention (TDQL) and high-dose group of the Chinese medicine composition of the present invention (TDQH). The ordinary feed group served as the blank control group (CON). The rats were continuously gavaged for 6 weeks. The specific intervention methods were as follows:

[0086] CON group: fed with normal diet and intragastrically administered with 5 ml / kg of distilled water once a day;

[0087] MOD group: fed with high-fat diet and intragastrically administered with 5 ml / kg of distilled water once a day;

[0088] TDQL group: fed with high-fat diet, and intragastric administration of 5 g / kg of the Chinese medicinal composition of the present invention once a day;

[0089] TDQH group: fed with high-fat diet, and intragastric administration of 20 g / kg of the Chinese medicinal composition of the present invention once a day;

[0090] MH group: The mice were fed with high-fat diet and given 0.30 g / kg of metformin hydrochloride tablets by oral gavage once a day.

[0091] 2.3 Experimental specimen collection

[0092] 2.3.1 Oral glucose tolerance test (OGTT): Rats were gavaged until the end of the sixth week, and all rats were fasted overnight but not water for 12 hours. Blood was collected from the tail vein in the early morning of the next day to detect FBG, which was set as the blood sugar value at 0 min. Then, 50% glucose was gavaged at a dose of 2 g / kg, and blood was collected from the tail vein at 30 min, 60 min, 90 min and 120 min after gavage to detect blood sugar at each time point. Finally, the corresponding curves were drawn according to the blood sugar levels of rats in each group at different times.

[0093] 2.3.2 At the end of the sixth week of gavage, all rats were fasted overnight but allowed to drink water for 8 hours. The next day, the fasting weight of the rats was measured, and the rats were anesthetized with 20% urethane solution. The anesthetized rats were fixed on the dissection table, and the abdominal aorta was selected for blood collection. The blood samples were stored at natural temperature for 2 hours, and then centrifuged at 3500r per minute for 15 minutes in a low-temperature high-speed centrifuge. The supernatant, i.e. serum, was taken out and stored in a -80℃ refrigerator for subsequent serological analysis. The small intestine was removed, rinsed with PBS, weighed and the data was recorded. Finally, the colon was cut into small sections with a knife, and the specimens were fixed in 4% paraformaldehyde solution and stored at 4℃ for subsequent pathology and qualitative detection of AHR and IL-22.

[0094] 2.4 IL-22 and IL-6 detection

[0095] The enzyme linked immunosorbent assay (ELISA) method was used to detect the concentration levels of IL-22 and IL-6 in rat serum. The detection of each serum index was carried out strictly in accordance with the operating instructions of the corresponding kit.

[0096] 2.5 HE staining of small intestine

[0097] (1) Specimen embedding and sectioning: The small intestinal tissue was fixed in 4% paraformaldehyde solution at 4°C for 24 hours and then cut into 3 μm slices.

[0098] (2) Dewaxing: Place the sections in xylene twice, each time for 5 min, then soak in 100% ethanol and 75% ethanol for 5 min, then rinse and dehydrate; finally soak in distilled water for 2 min.

[0099] (3) Staining: In an environment with a temperature of 37°C, use hematoxylin staining solution for 5 minutes and rinse with distilled water. Then place the specimen in differentiation solution for 3 minutes and rinse with tap water twice, each time for 2 minutes. Then place the specimen in eosin staining solution for 30 seconds and rinse with distilled water for 2-3 seconds, and dehydrate quickly.

[0100] (4) Dehydration and sealing: Soak the specimens in 75% and 100% ethanol for 2-3 seconds, then in 100% ethanol for 1 minute, and then transparentize with xylene twice, each time for 1 minute. Finally, seal with neutral gum and observe under a microscope.

[0101] 2.6 Immunohistochemistry

[0102] (1) Specimen preparation: The isolated small intestinal tissue was fixed with 10% formalin solution at 4°C overnight, and then cut into 3 μm slices. The tissue was dehydrated and embedded in paraffin according to the routine procedures.

[0103] (2) Antigen repair: Incubate the small intestinal slices in 3% hydrogen peroxide for 10 minutes at room temperature to eliminate the activity of endogenous peroxidase, and then rinse with PBS (PH7.4) solution 3 times, 5 minutes each time. Using the high-pressure heat repair method, place the small intestinal tissue slices in EDTA2Na (PH8.0) buffer, heat for 2 minutes, place at room temperature to cool naturally, and finally rinse the slices with PBS (PH7.4) solution for 3 minutes × 3 times. (3) Blocking: Dry the small intestinal slices with absorbent paper, then add 5% skim milk powder and block at 37°C for 1 hour.

[0104] (4) Incubation with primary antibody: IL22 and AHR antibodies were used as primary antibodies at dilution ratios of 1:200 and 1:250, respectively. 50 μl of primary antibody was added to each slice, and the negative control group was added with the corresponding dose of PBS solution. The slices were placed in a humidified box and incubated overnight at 4°C.

[0105] (5) Rewarming: Remove the slices that have been incubated overnight and place them at room temperature for 60 minutes to rewarm. Then rinse them three times with PBS (pH 7.4), each time for 5 minutes.

[0106] (6) Incubation with secondary antibody: Add 50 μl of rabbit secondary antibody-HRP polymer to each section, incubate at 37°C for 30 min, and rinse with PBS for 5 min each time for 3 times.

[0107] (7) Color development: shake off the PBS solution vigorously and wipe dry with absorbent paper; add 100ul DAB staining solution to each slice for color development and incubate for 5 minutes. Observe the staining effect under a microscope. If a brown-yellow color appears in the field of view, remove the slice and rinse with tap water to terminate the reaction.

[0108] (8) Restaining: Rinse the sections with tap water and then incubate with hematoxylin staining solution for 20 seconds; differentiate, rinse and blue again.

[0109] (9) Dehydration and sealing: The slices were placed in different concentrations of ethanol solution and soaked for 2 min. The slices were then soaked in anhydrous ethanol twice to dehydrate and become transparent. Finally, the slices were air-dried and then dripped with neutral gum to seal the slices.

[0110] (10) Microscopic examination: The Aipathwell analysis system was used for calculation. Three images were collected for each section, and the final scoring result was obtained based on the cell staining intensity and the number of positive cells.

[0111] 3. Experimental results

[0112] 3.1 Effect of the Chinese medicine composition of the present invention on the general condition of type 2 diabetic rats

[0113] The rats in the blank control group were healthy, flexible, with shiny and clean hair, normal drinking water and diet, moderate hardness of feces, and no deaths were observed during the experiment. The rats in the model group were obese, with little change in weight during the experiment, less activity, curled up and less breath, grayish-white and moist hair, significantly increased drinking water and food intake, slightly dry feces, and 2 rats died of hyperglycemia during the experiment. Compared with the model group, the hair of the rats in each intervention group was shinier and smoother, and their weight decreased during the experiment. They were relatively flexible, drank slightly less water and ate less than the model group, and had loose feces. During the experiment, due to improper gavage operation or other reasons, 2 rats in the low-dose group of the Chinese medicine composition of the present invention, 3 rats in the high-dose group of the Chinese medicine composition of the present invention, and 1 rat in the metformin group died. 3.2 Effect of the Chinese medicine composition of the present invention on blood glucose in rats with type 2 diabetes

[0114] In order to illustrate the effect of the Chinese medicine composition of the present invention on the glucose metabolism of insulin resistance in type 2 diabetes, the project team conducted an OGTT test on STZ-induced type 2 diabetic rats by drug gavage until the end of the sixth week, and compared the blood glucose levels of rats in each group at different time points after the OGTT test. The experimental results are as follows (see Table 5):

[0115] (1) Effect on fasting (0min) blood glucose: Compared with the control group, the fasting blood glucose of rats in the model group, the low-dose group, the high-dose group of the Chinese medicine composition of the present invention, and the metformin group increased significantly, and the difference was statistically significant (P<0.0001, P<0.0001, P<0.0001, P<0.0001); compared with the model group, the fasting blood glucose of each intervention group decreased, and the difference was statistically significant (P<0.0001, P<0.0001, P=0.001); there was no statistically significant difference between the low-dose group of the Chinese medicine composition of the present invention and the metformin group (P=0.293); there was a statistically significant difference between the high-dose group of the Chinese medicine composition of the present invention and the metformin group (P=0.025). There was no statistically significant difference between the low-dose group and the high-dose group of the Chinese medicine composition of the present invention (P=0.215).

[0116] (2) Effect on 30-min blood glucose: Compared with the control group, the fasting blood glucose of rats in the model group, the dosage group of the Chinese medicine composition of the present invention, the high dosage group and the metformin group was significantly increased, which was statistically significant (P<0.0001, P<0.0001, P<0.0001, P<0.0001); compared with the model group, the blood glucose of each drug-treated group was reduced, and the difference was statistically significant (P=0.001, P<0.0001, P=0.001); there was no significant statistical difference between the high dosage group of the Chinese medicine composition of the present invention and the metformin group (P=0.823, P=0.367); there was no significant statistical difference between the low dosage group and the high dosage group of the Chinese medicine composition of the present invention (P=0.276).

[0117] (3) Effect on 60-min blood glucose: Compared with the control group, the blood glucose of the rats in the model group, the low-dose and high-dose Chinese medicine composition of the present invention, and the metformin group was significantly increased, which was statistically significant (P<0.0001, P<0.0001, P<0.0001, P<0.0001); compared with the model group, the blood glucose of each drug-treated group was reduced, and the difference between the high-dose Chinese medicine composition of the present invention and the metformin group was statistically significant (P=0.005, P=0.021), and the difference between the low-dose Chinese medicine composition of the present invention and the metformin group was not statistically significant (P=0.069); there was no statistically significant difference between the low-dose and high-dose Chinese medicine composition of the present invention and the metformin group (P=0.635, P=0.480); there was no statistically significant difference between the low-dose and high-dose Chinese medicine composition of the present invention (P=0.258)

[0118] (4) Effect on 90-min blood sugar: Compared with the control group, the blood sugar of the rats in the model group, the low-dose and high-dose Chinese medicine composition of the present invention, and the metformin group increased significantly, with significant statistical significance (P<0.0001, P<0.0001, P<0.0001, P<0.0001); compared with the model group, the blood sugar of each drug-treated group decreased significantly, with statistically significant differences (P=0.003, P<0.0001, P<0.0001). There was no statistically significant difference between the low-dose and high-dose Chinese medicine composition of the present invention and the metformin group (P=0.414, P=0.336). There was no statistically significant difference between the low-dose and high-dose Chinese medicine composition of the present invention groups (P=0.091).

[0119] (5) Effect on 120-min blood glucose: Compared with the control group, the blood glucose of rats in the model group, the Chinese medicine composition of the present invention, the high-dose group and the metformin group increased significantly, with significant statistical significance (P<0.0001, P<0.0001, P<0.0001, P<0.0001). Compared with the model group, the blood glucose of each drug-treated group decreased, and the difference was statistically significant (P<0.0001, P<0.0001, P<0.0001). There was no statistically significant difference between the low-dose and high-dose groups of the Chinese medicine composition of the present invention and the metformin group (P=0.668, P=0.173). There was no statistically significant difference between the low-dose and high-dose groups of the Chinese medicine composition of the present invention (P=0.085).

[0120] Table 5 Comparison of blood glucose at each time point in OGTT test of rats in each group

[0121]

[0122] Note: Compared with the CON group, * P<0.05; compared with the MOD group, # P<0.05; compared with MH group, △ P<0.05.

[0123] 3.3 Effect of the Chinese medicine composition of the present invention on IL-22 and IL-6 in type 2 diabetic rats

[0124] Intestinal flora can activate AHR transcription factors, promote the secretion of anti-inflammatory factor IL-22, inhibit the secretion of pro-inflammatory factor IL-6, and thus inhibit metabolic inflammatory response. In order to clarify the effect of the Chinese medicine composition of the present invention on metabolic inflammation in rats with type 2 diabetes, blood was collected from the tail vein of rats with type 2 diabetes after 6 weeks of drug intervention, and ELISA was used to detect serum IL-22 and IL-6 in rats of each group. The results are shown in Table 6:

[0125] Effect on IL-22: Compared with the control group, the serum IL-22 of rats in the model group, the low-dose and high-dose Chinese medicine composition of the present invention, and the metformin group was significantly increased, indicating that the difference was statistically significant (P<0.0001, P<0.0001, P<0.0001, P<0.0001); compared with the model group, IL-22 in each intervention group was increased, proving that the difference was statistically significant (P<0.0001, P<0.0001, P<0.0001). There was no statistically significant difference between the low-dose group of the Chinese medicine composition of the present invention and the high-dose group of the Chinese medicine composition of the present invention and the metformin group (P=0.064, P=0.621). There was no statistically significant difference between the low-dose group and the high-dose group of the Chinese medicine composition of the present invention (P=0.16).

[0126] Effect on IL-6: Compared with the control group, the serum IL-6 of rats in the model group, the low-dose and high-dose Chinese medicine composition of the present invention, and the metformin group increased, and the results showed that the difference was statistically significant (P<0.0001, P<0.0001, P<0.0001, P<0.0001); compared with the model group, each intervention group showed an obvious trend of IL-6 reduction, and the results showed that the difference was statistically significant (P<0.0001, P<0.0001, P<0.0001); compared with the metformin group, the low-dose group and the high-dose group of the Chinese medicine composition of the present invention had higher IL-6 values ​​in the low-dose group, and the difference was statistically significant (P<0.0001), and the high-dose group had slightly lower IL-6, but the difference was not statistically significant (P=0.453). Compared with the low-dose group and the high-dose group of the Chinese medicine composition of the present invention, the difference was statistically significant (P<0.0001).

[0127] Table 6 Comparison of IL-22 and IL-6 in rats of each group (n=5)

[0128]

[0129] Note: Compared with CON, * P<0.05; compared with MOD, # P<0.05; compared with MH, △ P<0.05; compared with TDQL, ※ P<0.05.

[0130] 3.4 Effect of the Chinese medicine composition of the present invention on small intestinal tissue of rats with type 2 diabetes

[0131] (1) HE staining (such as Figure 4) as shown in the figure: the normal control group rats had neat and dense arrangement of the superficial layer of the small intestinal mucosa, densely arranged intestinal glands in the lamina propria, small focal infiltration of inflammatory cells and eosinophils and crypt inflammation in the intestinal epithelial lamina propria, focal degeneration of the intestinal epithelium, small focal atrophy of the villi, and crypt hyperplasia. Model group: partial infiltration of inflammatory cells in the intestinal epithelial lamina propria, focal degeneration of the intestinal epithelium, focal cavitation of the intestinal epithelium, partial atrophy of the villi, crypt inflammation and crypt abscess formation, and crypt hyperplasia. The intestinal epithelium and crypt focus regeneration were active in the low-dose and high-dose groups of the Chinese medicine composition of the present invention and the metformin group, the mucosa was atrophied, the crypt glands lost parallel arrangement and branches appeared, and the crypt glands were shortened.

[0132] (2) Immunohistochemical test results (such as Figure 5 ): The small intestinal cells of the control group rats were lightly stained, and only partially stained, or even not stained; the model group showed local cell staining, which was light brown; compared with the model group, the cells of the low-dose group, high-dose group and metformin group of the Chinese medicine composition of the present invention were obviously brown-yellow in color, and the crypt glands were darker in color. The results suggest that AHR and IL-22 were less expressed in the small intestinal tissue of the control group rats, and even showed negative expression; AHR and IL-22 were mainly weakly positive in the model group rats, and villus atrophy and intestinal epithelial degeneration of the small intestinal tissue were observed; the expression of the low-dose group, high-dose group and metformin group of the Chinese medicine composition of the present invention was significantly increased compared with the model group, mostly showing moderate positive expression or even strong positive expression, and the crypt glands were observed to regenerate and lose parallel arrangement and develop to the healing stage.

[0133] The results showed that the Chinese medicine composition of the present invention can inhibit the level of pro-inflammatory factor IL-6 in the serum of type 2 diabetic rats (P<0.05) and increase the level of anti-inflammatory factor IL-22 (P<0.05). In the small intestinal tissue of type 2 diabetic rats, the Chinese medicine composition of the present invention can alleviate inflammatory infiltration; the results of immunohistochemical detection showed that the Chinese medicine composition of the present invention can increase the expression levels of AHR and IL-22 in small intestinal tissue.

Claims

1. A hypoglycemic Chinese medicine composition for inhibiting metabolic inflammation, characterized in that: The invention is prepared from the following raw materials in parts by weight: 20-40 parts of perilla, 5-7 parts of coptis root, 10-20 parts of ginseng, 10-20 parts of tuckahoe, 10-20 parts of stir-fried atractylodes, 5-7 parts of roasted licorice, 10-20 parts of patchouli, 8-10 parts of costusroot, 12-22 parts of kudzu root, 8-10 parts of bombyx batryticatus, 10-20 parts of euphorbia pulex, 10-20 parts of mulberry branch and 2-5 parts of cinnamon bark.

2. The hypoglycemic Chinese medicine composition for inhibiting metabolic inflammation according to claim 1, characterized in that: The weight of the raw materials is: 30g of Perilla frutescens, 6g of Coptis chinensis, 15g of Ginseng, 15g of Poria, 15g of Stir-fried Atractylodes macrocephala, 6g of Rhizoma Licorice, 15g of Patchouli, 9g of Aucklandia lappa, 18g of Pueraria root, 9g of Bombyx batryticatus, 15g of Psoralea corylifolia, 15g of Morus alba, and 3g of Cinnamon bark.

3. The method for preparing the hypoglycemic Chinese medicine composition for inhibiting metabolic inflammation according to claim 1 or 2, characterized in that: Weigh the medicinal materials of Perilla frutescens, Coptis chinensis, Ginseng, Poria cocos, stir-fried Atractylodes macrocephala, roasted Licorice root, Patchouli, Aucklandia lappa, Pueraria root, Bombyx batryticatus, Psoralea corylifolia, Morus alba, and Cinnamomum cassia, soak the above medicinal materials in 5 times the weight of pure water for 2 hours, boil them with high heat at 150-220° C., then decocted with low heat at 80-120° C. for 30 minutes, filter and collect the decoction; add 2 times the weight of pure water to the filter residue and decoct for 20 minutes, filter and collect the decoction; mix the two decoctions, and concentrate them to a Chinese medicine concentration of 2.5 g / ml to obtain the Chinese medicine composition of the present invention.

4. The hypoglycemic Chinese medicine composition for inhibiting metabolic inflammation according to claim 1, characterized in that: The pharmaceutical preparation is made from the following raw materials in parts by weight: 20-40 parts of perilla, 5-7 parts of coptis root, 10-20 parts of ginseng, 10-20 parts of poria, 10-20 parts of stir-fried atractylodes, 5-7 parts of roasted licorice, 10-20 parts of patchouli, 8-10 parts of costus root, 12-22 parts of kudzu root, 8-10 parts of bombyx batryticatus, 10-20 parts of euphorbia pulex, 10-20 parts of mulberry twig, 2-5 parts of cinnamon bark, and pharmaceutically acceptable excipients.

5. The hypoglycemic Chinese medicinal composition for inhibiting metabolic inflammation according to claim 4, characterized in that: The pharmaceutical preparations include injections, oral liquids, pills, powders, pastes, tablets, granules, powders or capsules.

Images