| The first part:Intervention of Scrophularia radix-Atractylodes lancea on Type 2Diabetic rats1 Objectives and Significance:In recent years,Type 2 Diabetes Mellitus(T2DM)has become a threat to human life and health after tumor and cardiovascular and cerebrovascular diseases.Shi Jinmo Dui Yao believes that Scrophularia radix-Atractylodes lancea has a good hypoglycemic effect.In this part,animal experiments confirm the hypoglycemic effect of Scrophularia radix-Atractylodes lancea.2 Methods:After 8 weeks of high-fat diet,the rats were fasted for 12 hours and then injected with small dose STZ(30mg · kg-1)intraperitoneally.The rats with fasting blood glucose ? 16.7mmol · L-1 were selected as the qualified type 2 diabetic rats model.The animal model was randomly divided into Model Group?Positive Drug Group,TCMlow-dose Group,TCMmiddle-dose Group,TCMhigh-dose Group,and Normal Control Group.Rats were administrated by gavage for 2 weeks,Rats in each group were fed with normal diet instead of normal saline.Fasting blood glucose and 2 hours postprandial blood glucose were monitored weekly before and during administration.The data were collected and analyzed by SPSS 22.3 Result:The results showed that: compared with the normal control group,the FBG and2 h PG of type 2 diabetic rats increased significantly,and the difference between the groups was statistically significant(P<0.05).Compared with the model group,the FBG and 2h PG of the positive drug group and the traditional Chinese medicine group decreased,but the difference between the groups was not statistically significant(P>0.05).4 Conclusions:(1)The T2 DM rat model can be successfully established by high-fat diet for 8weeks combined with intraperitoneal injection of 1% streptozotocin solution30 mg · kg-1.(2)The hypoglycemic effects of FBG and 2h PG in rats were not significant after two weeks of administration of Scrophularia radix-Atractylodes lancea alone.The second part: The increase hypoglycemic effect of Scrophularia radixAtractylodes lancea combined with Metformin on T2 DM rats1 Objectives and Significance:In the first part of the study,it was found that the hypoglycemic effect of Scrophularia radix-Atractylodes lancea for two weeks on FBG and 2h PG was not significant.In order to avoid the failure of the experiment,this part adjusted the experimental scheme on the basis of the first part to explore the hypoglycemic and synergistic effect of Scrophularia radix-Atractylodes lancea combined with Metformin on T2 DM rats.2 Methods:On the basis of the first part,the rats in each group of TCM were administrated with Metformin for 2 weeks,blood glucose was monitored and statistically analyzed.The therapeutic effect of Scrophularia radix-Atractylodes lancea on reducing blood glucose was confirmed.After 2 weeks,the rats were fasted for 12 hours,the serum AST?ALT?GGT?Urea?TG?CHO?LDL-C?HDL-C levels of rats were measured.After the blood was taken,the liver and kidney were taken out and weighed,and the data were sorted out.SPSS 22 was used to analyze the data.3 Result:(1)Blood glucose monitoring after administration of Scrophularia radixAtractylodes lancea combined with Metformin,we found:compared with the Positive Drug Group,the FBG in the Positive+TCMmedium-dose Group decreased significantly(P<0.05),Compared with the model group,the 2h PG in the Positive+TCMlow-dose Group and the Positive+TCMmedium-dose Group decreased significantly(P<0.05),while the blood glucose in the positive drug group was not significant(P<0.05)It was proved that the effect of Scrophularia radix-Atractylodes lancea was good;(2)Compared with the Blank Control Group,the intake?water drinking?liver weight index?kidney weight index?kidney weight index of rats in the other groups increased significantly(P<0.05),and body weight decreased significantly(P < 0.05),Compared with the Model group and the Positive Drug Group,the amount of food and water in the Positive+TCM Group decreased(P<0.05).Compared with the Model Group,the liver weight index in the Positive+TCMhigh-dose Group increased significantly(P<0.05).Compared with the Positive Drug Group,the kidney weight in the Positive+TCMmedium-dose Group and the Positive+TCMhigh-dose Group decreased(P < 0.05);(3)The correlation analysis showed that FBG was positively correlated with water intake,abdominal circumference,liver weight,liver weight index,kidney weight index,serum ALT level,serum GGT level,serum Urea level and negatively correlated with body mass(P<0.05);2h PG was positively correlated with food intake?water intake?liver weight index?kidney weight index?ALT level?AST level?GGT level and Urea level(P < 0.05).4 Conclusions:(1)Scrophularia radix-Atractylodes lancea has significant hypoglycemic effect.(2)Scrophularia radix-Atractylodes lancea can improve the general condition(such as intake?water drinking?body weight?liver and kidney weight index)and biochemical indexes(such as FBG?2h PG?LDL-C?ALT?Urea?etc.)of T2 DM rats;(3)Blood glucose was positively correlated with water intake ? abdominal circumference?liver weight?liver weight index?kidney weight index?ALT level?GGT level ? Urea level,and negatively correlated with body weight;2h PG was positively correlated with food intake?water intake?liver weight index?kidney weight index?ALT level?AST level?GGT level and Urea level.The third part : the effect of Scrophularia radix-Atractylodes lancea on the structure of intestinal flora in Type 2 Diabetic rats1 Objectives and Significance:At present,most studies have confirmed the phenomenon of flora imbalance in T2 DM patients.Through the second part of the study,we found that Scrophularia radix-Atractylodes lancea combined with Metformin has a hypoglycemic and synergistic effect.In order to explore whether Scrophularia radix-Atractylodes lancea has changed the structure of intestinal flora at the same time of hypoglycemic,we carried out the third part of the study.2 Methods:The fresh fecal samples of the rats were collected and stored in a closed fecal storage box in a-80?refrigerator,and then sent to Shanghai paisenno Biotechnology Co,Ltd.for gene sequencing of intestinal flora by 16 S r DNA high-throughput sequencing technology.3 Result:(1)Compared with the normal rats,the relative abundances of Blautia ?Allobaculum and Phascolarctobacterium in the Model Group were significantly higher(P<0.05),and the relative abundances of Turicibacter?Desulfovibrio in the Model Group were significantly lower(P<0.05),Compared with the Model Group,the relative abundance of Blautia in the Positive Drug Group and the Positive+ TCM Group decreased,but the difference was not statistically significant(P>0.05).The relative abundance of Tricibacter in the Positive Grug Group and the Positive+TCM Group increased significantly(P < 0.05),The relative abundance of Roseburia decreased significantly(P<0.05);(2)Pediococcus ?Corynebacterium and Odoribacter were found in the Blank Control Group;Blautia was found in the Model Group;Sutterella and Parabacteroides were found in the Positive+TCMmedium-dose Group;and Sutterella and Parabacteroides were found in the Positive+TCMmedium-dose Group,Bifidobacterium and Phascolarctobacterium were found in the Positive+TCMhigh-dose Group;(3)The results showed that there was a positive correlation among the beneficial bacteria,such as: Lactobacillus?Sutterella?Parabacteroides,and the foreign bacteria,such as Tricibacter ? Blautia ? Bifidobacterium and Phascolarctobacterium ?Desulfovibrio?Pediococcus?Corynebacterium?Odoribacter and Allobaculum,and a negative correlation between beneficial bacteria and foreign bacteria(all P<0.05);(4)The PCA analysis showed that FBG and 2h PG were the main factors driving the intestinal community variation,while blautia,Pediococcus and Roseburia were positive related to FBG and 2h PG,negative related to FBG and 2h PG(all P < 0.05).4 Conclusions:(1)The results showed that the relative abundances of Blautia?Allobaculum and Phascolarctobacterium in T2 DM rats were significantly higher than those in normal rats,and the relative abundances of Tricibacter and Desulfovibrio were significantly lower;Metformin and Scrophularia radix-Atractylodes lancea also affected structure of T2 DM flora,The results showed that Metformin could increase the relative abundance of Tricibacter and decrease the relative abundance of Blautia;Scrophularia radix-Atractylodes lancea significantly reduced the relative abundance of Roseburia;(2)It was found that the biomarkers of normal rats were Pediococcus ?Corynebacterium and Odoribacter.The biomarkers of type 2 diabetic rats were Blautia.The biomarkers of Positive+TCMmedium-dose were Sutterella and Paracharacteroids,The biomarkers of Positive+TCMhigh-dose were Bifidobacterium and Phascolarctobacterium.(3)It was found that FBG and 2h PG were the main factors driving the variation of intestinal community,while Blautia was positively correlated with FBG and2 h PG,Pediococcus was nagetively correlated with FBG and 2h PG,Roseburia was positively correlated with 2h PG,they plays an important role in driving the variation of intestinal community.These three genera are closely related to other genera and other clinical indicators. |
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