Based On The Homology Of Liver And Kidney To Explore The Research Of Qidi Tangshen Recipe On Fibrosis And Bile Acid In DN Mice

Research background and purposeDiabetic nephropathy is one of the common clinical diseases,which causes heavy economic burden to the society.Renal fibrosis is one of the important pathological manifestations of diabetic nephropathy.Previous studies have believed that bile acids are closely related to glycolipid metabolism.However,in recent years,scholars have found that there are abundant bile acid receptors in the kidney,and there is evidence that bile acid pathway is related to fibrosis.Homology of liver and kidney is an important concept in traditional Chinese medicine,which has important guiding significance for the treatment of diabetic renal disease.It is not clear how the bile acid-renal fibrosis pathway is related to the homology of liver and kidney in traditional Chinese medicine.The purpose of this study is to explain the material basis of liver and kidney homology by analyzing the correlation between the levels of bile acid and renal fibrosis indexes and the homologous syndrome elements of liver and kidney,so as to enrich the scientific connotation of liver and kidney homology.Albuminuria is one of the important clinical manifestations of diabetic nephropathy.Previous studies suggested that Qiditangshen recipe could reduce the level of albuminuria in diabetic nephropathy rat model,but the specific mechanism remains to be further studied.It has not been reported whether Qiditangshen decoction can reduce the renal fibrosis status of diabetic nephropathy and diabetic nephropathy proteinuria through bile acid pathway.This study intends to reveal the mechanism of Qiditangshen prescription in the treatment of diabetic nephropathy proteinuria through the intervention of Qiditangshen prescription in DB/DB diabetic nephropathy mouse model.The research methodsThe clinical partThrough the collection of outpatient and inpatient patients from Dongzhimen Hospital of Beijing University of Chinese Medicine as well as 83 patients with diabetic nephropathy and diabetes in the physical examination of nearby communities,the general basic information,urine and serum samples of the patients were collected,and the relevant symptoms were collected and scored according to the TCM syndrome element scale of our team.According to the microalbumin/urinary creatinine(ACR)value,the patients were divided into two groups:simple diabetes mellitus(DM)group and diabetic nephropathy(DKD)group.The levels of urinary bile acid,p-Ⅲ COL,Pa-COL,LN,HA and serum TGF-β1 were detected by ELISA.The expression levels of TGF-β1 in liver deficiency syndrome and kidney deficiency syndrome related syndromes,bile acid,p-Ⅲ COL,AD-COL,LN,HA and serum were compared between DM group and DKD group.To investigate the correlation between the expression levels of bile acid,p-Ⅲ COL,AL-COL,LN,HA and TGF-β1 and the symptoms of liver deficiency syndrome and kidney deficiency syndrome.The experiment partDiabetic nephropathy model was prepared by feeding 8-week-old obese and spontaneous type 2 diabetic DB/DB male mice(grade SPF),and 30 8-week-old C57BL/6J male mice were purchased as control group.Mice aged 14 weeks were divided into normal group(n=20)and model group(n=45)by random grouping.Normal mice were divided into normal group and normal administration group,and the model group was divided into model group,western medicine group and Chinese medicine group.Each group was given specific drugs prepared with normal saline,once a day,with the same volume of gavage fluid,and the specific dose was based on the effective dose in previous studies.During drug administration,all mice were weighed every 2 weeks,blood was collected from tail tip every 2 weeks for blood glucose measurement,and urine was collected in metabolic cages for 8 hours every 4 weeks.Blood creatinine,triglyceride,cholesterol,alanine aminotransferase and aspartate aminotransferase were detected at the 12th weekend after administration.He and Masson staining light microscopy was used to observe the renal structure and fibrosis staining changes.Ultra-micro pathological structure of the kidney was observed by electron microscopy.The changes of renal fibrosis indexes were observed by immunohistochemistry.On this basis,the expression of FXR in mouse kidney tissue was detected by Western-blot method to preliminarily explore the mechanism of bile acid regulating renal fibrosis.The resultsThe clinical partCompared with the diabetic group,ALT,AST,urea nitrogen.EGFR,creatinine and total cholesterol in the diabetic nephropathy group were statistically significant(P<0.05),and there was no statistical difference in other indicators(P>0.05).Compared with diabetic group,there were no significant differences in liver fibrosis,bile acid and TGF-β 1 in diabetic nephropathy group(P>0.05).Compared with the diabetic group,the score of liver and blood deficiency in the diabetic nephropathy group was significantly higher(P<0.05),and other syndromes had no significant statistical significance.The correlation analysis of bile acid,TGF-β1,p-Ⅲ COL,Di-COL,LN and HA in patients with diabetic kidney disease was conducted,respectively,with liver deficiency,kidney deficiency,liver and blood deficiency,liver Yin deficiency,kidney Yin deficiency,kidney Yang deficiency,kidney essence deficiency,liver and kidney deficiency and related symptoms.The results showed that bile acid was positively correlated with liver and blood deficiency,and the difference was statistically significant.HA was positively correlated with renal essence deficiency score and liver and kidney deficiency score,and the difference was statistically significant.Collagen 3 was positively correlated with renal essence deficiency score and liver and kidney deficiency score,and LN was negatively correlated with visual object failure score(P<0.05),TGF-β 1 was positively correlated with the visual fainting score(P<0.05),bile acid had a negative correlation with visual coma score(P<0.05),LN was negatively correlated with lumbar and knee cold pain scores(P<0.05),HA was positively correlated with helix shrinkage score(P<0.05),HA was negatively correlated with urine shortness,lumbar and knee cold pain(P<0.05),bile acid was positively correlated with depression score(P<0.05).The experiment partCompared with the normal group,there was no significant difference in biochemical parameters between the normal administration group(P>0.05),the decrease of aspartate aminotransferase in western medicine group was statistically significant(P<0.05).Compared with the model group,the decrease of aspartate aminotransferase in the western medicine group was statistically significant(P<0.05),the decrease of alanine aminotransferase in TCM group was statistically significant(P<0.05).Compared with the normal group,there were no differences in blood glucose and body weight changes in the normal administration group(P>0.05),there were statistical differences in model group,western medicine group and Chinese medicine group(P<0.05).Compared with model group,there was no significant difference in blood glucose and body weight between western medicine group and Chinese medicine group(P>0.05).Compared with the normal group,the levels of urinary microalbumin in the model group,western medicine group and Chinese medicine group were all increased before gavage,and also increased three times after gavage,and the difference was statistically significant(P<0.05).Compared with the model group,there was no difference between the western medicine group and the Chinese medicine group before gavage,and the three tests after gavage were all decreased,and the difference was statistically significant(P<0.05).Compared with the normal group,TGF-β1 Smad2,Smad3 and α-SMA levels in the model group were significantly different(P<0.05),the levels of TGF-β1,Smad2,Smad3 and α-SMA in western medicine group and Chinese medicine group were significantly different compared with model group(P<0.05).Masson staining showed that,compared with the normal group,the interstitial fibrosis and hyperplasia were more obvious.However,for the western medicine group and the Chinese medicine group,the mesangial matrix hyperplasia was not obvious,and the basement membrane of the glomerulus was not significantly thickened.In particular,the fibrin deposition in the renal interstitium was significantly improved compared with the model group.Bile acid composition analysis showed that compared with the normal group,the levels of TCA and T-β-MCA in the model group were increased,and the difference was statistically significant(P<0.05).Compared with model group,the levels of β-MCA and T-β-MCA and TCA in TCM group were decreased,and the difference was statistically significant(P<0.05).There was no statistical difference in other indicators.WB method was used to detect the level of renal FXR,and there was no significant statistical difference among the groups.Correlation analysis of bile acid components and fibrosis factors showed that T_BAS,CDCA,α-MCA,TCDCA,T-β-MCA were positively correlated with Smad3,T-β-MCA was positively correlated with TGF-β1,and T-β-MCA and TCA were positively correlated with Smad2.conclusionFor patients with diabetic kidney disease,bile acid and fibrosis factors are correlated with the syndrome of liver deficiency and kidney deficiency,which enriches the scientific connotation of the theory of homology between liver and kidney.Qiditangshen prescription can regulate the secretion of bile acid,improve renal fibrosis,and reduce diabetic nephropathy proteinuria.