Study On The Mechanism Of Anomie Of Syndrome Differentiation Of Diabetes And The Combination Of Astragalus And Pueraria Lobata Root In Regulating Insulin Secretion

I.Literature ResearchPurpose:To clarify the problems of traditional Chinese medicine(TCM)syndrome differentiation in diabetes mellitus(DM),in order to provide methods and ideas for the standardization of TCM dialectics.Material and method:Collection of syndrome differentiation in authoritative literature,such as curative Effect of TCM syndrome diagnosis standards,Guidelines for the Research on new TCM drugs,Guideline for TCM Diabetes Prevention and Treatment,the Internal Medicine of TCM.Computer retrieval of DM traditional Chinese medicine syndrome differentiation documents in core journals included in China How Net and Wan Fang database from 2014 to 2018,and collated and analyzed.It clearly reveals different kinds of problems of TCM syndrome differentiation between authoritative literature and academia about DM.And then provide methods and ideas for the standardization of TCM syndrome differentiation of DM.Results: 1.In the comparison of authoritative literature,there are great differences between Standards and Principles in syndrome differentiation of DM.There are 9 syndromes:extreme heat with Yin deficiency,gastric dryness fluid injury,deficiency of kidney Yin,Yin-Yang deficiency,Yin deficiency and Yang float,Damp heat obstruct the spleen,qi-Yin deficiency,Yin-Yang deficiency and stagnation of blood and water,blood stasis in collaterals.Five syndromes in the Guide all inherits the Standards and Principles.In addition,there are two special syndromes in the Internal Medicine of TCM: Lung heat fluid injury,qi-Yin deficiency.The authoritative literature involved 11 syndromes.The Principles and Guidelines are currently the standard of general clinical reference,and it is reasonable to use them as authoritative literature in this study.The number of references in TCM Internal Medicine is medium,while the research on Standards is rare.2.Taking T2 DM as an example to investigate the clinical syndrome differentiation of TCM.It could divides into main syndrome differentiation and concurrent syndrome differentiation.The main syndromes include extreme heat with Yin deficiency,gastric dryness fluid injury,deficiency of kidney Yin,Yin-Yang deficiency,Yin deficiency and Yang float,qi-Yin deficiency,Yin-Yang deficiency and stagnation of blood and water,blood stasis in collaterals,Lung heat fluid injury,phlegm-heat syndrome,a total of 10 syndromes,and 27 new syndromes.The types of proofs with frequency in the top 10,Spleen deficiency and dampness,Liver depression and qi Stagnation,blood stasis due to qi deficiency,Liver-stomach heat stagnation were not included in the authoritative literature.The concurrent syndromes include phlegm turbidity and blood stasis syndromes in authoritative literature,which increase the four syndromes Stasis,turbid stasis,blood stasis due to qi stagnation,qi-Yin deficiency.The results show that there are three patterns of syndrome differentiation in clinic: full compliance with authoritative literature,be flexible on the basis of authoritative literature,flexible introduction of new syndromes.3.Taking qi-Yin deficiency syndrome and extreme heat with Yin deficiency syndrome as examples to investigate the composition of symptoms.The results showed that there were 59 symptoms of Qi-Yin deficiency syndrome,including all the symptoms of Qi-Yin deficiency syndrome in authoritative literature,and 34 new symptoms;extreme heat with Yin deficiency syndrome had 40 symptoms,including all the symptoms of Qi-Yin deficiency syndrome in authoritative literature(except coating thin yellow),and 18 new symptoms.Among the newly added symptoms of the two syndromes,some have the same symptoms and different names,some have no direct relationship with the disease,and some are tedious and repetitive.This increase in non-standard symptoms may be the cause of virtual syndrome.Conclusion: 1.The standards for syndrome differentiation are complex,diverse and different from each other in TCM for DM,lacking binding force in clinical implementation.2.Clinical syndrome differentiation of Type 2 Diabetes Mellitus in TCM is varied and discrete.The symptoms of various syndromes are complicated,which do not correspond to the symptoms,and the symptoms are not standardized.The newly added syndromes and symptoms lack objectivity and scientificity.3.Coordination and Integration of relevant departments,pay attention to standardization of symptoms in DM,unify syndrome type Name and standardize syndrome differentiation,research on prospective standards of syndrome differentiation of DM.These are ideas and methods to establish syndrome differentiation standard.II.Experimental ResearchPurpose:To clarify the effect of effective components compatibility of Astragalus and Kudzu on insulin secretion in diabetic rats.By studying the lipid metabolism,inflammatory reaction and phosphatidylinositol-3-kinase/protein kinase B-2(PI3K/AKT)signal transduction pathway of pancreas in diabetic rat model,to explore the mechanism of effective components compatibility of Astragalus and Kudzu on insulin secretion.Material and method:According to blood sugar factors and factorial experimental design scheme,146 SPF SD male rats were randomly divided into 10 groups.Which namely(1)normal group,(2)model group,(3)jin-qi-jiang-tang tablet group(1.47 g·kg-1),(4)Astragalus flavonoid group(0.039 g·kg-1),(5)Astragalus saponin group(0.104 g·kg-1),(6)Kudzu flavonoid group(0.036 g·kg-1),(7)Astragalus flavonoid and Astragalus saponin group(0.143 g·kg-1),(8)Astragalus flavonoid and Kudzu flavonoid group(0.075 g·kg-1),(9)Astragalus saponin and Kudzu flavonoid group(0.140 g·kg-1),(10)Astragalus flavonoid and Astragalus saponin and Kudzu flavonoid group(0.179 g·kg-1).In addition to the normal group,the other groups were induced by streptozocin(STZ,46 mg·kg-1)to build experimental diabetic rat model.Blood samples were taken from caudal vein to detect blood sugar in rats on 7d,random blood sugar(?16.8 mmol·L-1)was regarded as an experimental diabetic model rat.Model manufacturing on the same day as intragastric administration,normal group and model group were given the same amount of distilled water 10 m L·kg-1,each group was given 30 days continuously.The level of blood glucose was detected by using glucose meter on given drug 0d,7d,30 d respectively.Serum cholesterol(CHO)and triglyceride(TG)levels were measured by biochemical analyzer.Enzyme-linked immunosorbent assay(ELISA)was used to test the level of insulin(INS),adiponectin(APN),interleukin-12(IL-12),interleukin-15(IL-15),tumor necrosis factor(TNF-?)levels in the pancreas.The expression level of adiponectin receptor1(Adipo R1),AMP-activated protein kinase(AMPK),peroxisome proliferator-activated receptor ?(PPAR?),p38 mitogen-activated protein kinase(p38MAPK),insulin receptor substrate 2(IRS2),phosphatidylinositol-3-kinase(PI3K),protein kinase B-2(Akt2)and glycogen synthase kinase 3?(Gsk3?)m RNA in pancreas were assayed by Quantitative Real-Time PCR(q PCR).Statistical software SPSS 17.0 was used to analyze the experimental data.Results: 1.The effect of compatibility of effective components of Astragalus and Kudzu on pharmacodynamic indexes of diabetic rats.1.1 The effect on blood sugar in diabetic rats Before modeling,there was no significant difference in blood glucose among all experimental groups(P>0.05).After 7 days,compared with the normal group,the blood glucose level was increased significantly in groups of diabetic models(P<0.05).After 30 days,compared with the model group,only Astragalus flavonoid could significantly reduced the blood glucose level of DM model rats.Astragalus saponin and Kudzu flavonoid had no significant effect,and the blood glucose of the component compatibility group had no significant change.1.2 The effect on blood lipid of DM model rats Compared with the normal group,the serum levels of TG and CHO in the model group increased significantly(P<0.05).For serum TG level,besides the combination of three components,the compatibility of effective components of Astragalus and Kudzu had significant effects(P<0.05).For the serum CHO level,the compatibility of effective components of Astragalus and Kudzu had significant effects(P<0.05).1.3 The effect of insulin level on diabetic rats Compared with the normal group,the level of INS decreased significantly in the model group(P<0.05).Compared with model group,the level of INS increased in Astragalus flavonoid group and Kudzu flavonoid group(P<0.05).Astragalus saponin had no significant effect on the level of INS.Although there was interaction between Astragalus saponin andAstragalus flavonoid,it could not significantly increased the level of INS in model rats.The level of INS in the remaining compatibility group increased significantly,and there was interaction.The compatibility of Astragalus flavonoid and Kudzu flavonoid was the best combination to promote INS level,and was superior to the component used alone.2.Study on the pharmacodynamic mechanism of the compatibility of effective components of Astragalus and Kudzu in regulating insulin secretion in diabetic rats.2.1 The effect on the expression of lipid metabolism-related factors in pancreas Compared with the normal group,the level of APN decreased,the genes expression of Adipo R1,AMPK and PPAP? decreased(P<0.05).Astragalus flavonoid,Astragalus saponin,Kudzu flavonoid could increased the expression of Adipo R1 m RNA and PPAP? m RNA(P<0.05),but had no significant effect on the expression of APN.Only Astragalus flavonoid could significantly regulated the expression of AMPK m RNA in pancreas.The compatibility of Astragalus flavonoid and Astragalus saponin,Astragalus flavonoid and Kudzu flavonoid,three components could significantly increased the level of APN,but there was no significant interaction between the compatibility.The compatibility of Astragalus flavonoid and Astragalus saponin could synergistically regulated the expression of Adipo R1 m RNA in pancreas.The compatibility of Astragalus flavonoid and Kudzu flavonoid could synergistically regulated the expression of Adipo R1 m RNA and PPAP? m RNA in pancreas.The combination of the three components could coordinately regulated the expression of PPAP? m RNA in pancreas.The compatibility of Astragalus flavonoid and Kudzu flavonoid was the best combination to improve the expression of Adipo R1 m RNA.The combination of three components was the best combination to regulate the expression of PPAP? m RNA.2.2 The effects on expression of inflammatory index in pancreas Compared with normal group,the expression of IL-12,IL-15,TNF-? and p38 MAPK m RNA increased in the pancreas of the model group rats(P<0.05).There was significant interaction between effective components compatibility of Astragalus and Kudzu in the regulation of IL-15,TNF-?,p38 MAPK m RNA in pancreas.There was significant interaction between Astragalus flavonoid and Kudzu flavonoid in the regulation of IL-12(P<0.05).The compatibility of Astragalus flavonoid and Kudzu flavonoid was the best combination to reduce the expression of IL-12,IL-15 and p38 MAPK m RNA.The compatibility of Astragalus flavonoid and Astragalus saponin was the best combination to reduce the TNF-? level.2.3 The effect of PI3K/Akt signal transduction pathway in pancreas Compared with normal group,the expression of IRS2,PI3 K and Akt2 m RNA decreased,the expression of Gsk3? m RNA increased in pancreas(P<0.05).There was significant interaction between effective components compatibility of Astragalus and Kudzu in the regulation of IRS2 m RNA and Gsk3? m RNA in pancreas.There was significant interaction between Astragalus flavonoid and Kudzu flavonoid in the regulation of PI3 K m RNA in pancreas.The compatibility of Astragalus flavonoid and Astragalus saponin and the combination of three components had significant interaction on the regulation of Akt2 m RNA in pancreas.The compatibility of Astragalus flavonoid and Astragalus saponin was the best combination to improve the expression of IRS2 m RNA and Gsk3? m RNA in pancreas.The compatibility of Astragalus flavonoid and Kudzu flavonoid was the best combination to improve the expression of PI3 K m RNA in pancreas.The compatibility of three components was the best combination to regulate the expression of Akt2 m RNA in pancreas.Conclusion: 1.The compatibility of effective components of Astragalus and Kudzu could promote the secretion of INS in different degrees.The compatibility of Astragalus flavonoid and Kudzu flavonoid was the best combination to promote INS secretion.2.The compatibility of effective components of Astragalus and Kudzu could promote the secretion of INS,the mechanism may be related to alleviate toxicity of pancreatic lipid,may also associated with inhibit inflammation and regulate IRS/PI3K/Akt pathway of the pancreas.3.Astragalus flavonoid play a major role in the compatibility of components.The compatibility of components had both consistency and emphasis on the target of regulation.The regulatory effects of compatibility on target are synergistic,antagonistic or individually used.