Based On "renal Collateral Disease", The Effects Of Qiangshen Mixture On The Number And Morphological Structure Of Renal Interstitial TCs In IgAN Mice Were Investigate

Purpose:The number and morphology of TCs in the renal interstitium of IgAN mice were investIgAted compared with normal mice.To evaluate the curative effect of strengthening kidney mixture based on the principle of "tonIFying kidney and llFting collaterals,eliminating stasis and clearing collaterals" on IgAN mice and its protective and repairing effect on TCs in renal interstitium.MateriaL and method:1.To observe the changes of renal interstitial TCs in IgAN miceTwenty-four 6-week-old clean BAlB/c mice,male,were randomly divided into two groups:normal group(12 mice)and model group(12 mice).IgAN mice model was established by BSA+lPS+CC14 method.At the end of the 9th week,the mesangial cells and matrix of glomerulus were collected and observed by light microscope.IgA deposition in mesangial region was observed by immunofluorescence method.After the successful modeling,kidney samples were collected,and the changes of TCs in renal interstitial of IgAN mice were observed by immunofluorescence double labeling staining.The ultrastructural changes of renal interstitial TCs and its Telopodes(TPs)in IgAN mice were observed by transmission electron microscopy.2.To explore the protective and repairing effect of strengthening kidney Mixture on renal interstitial TCs in IgAN miceForty-eight male 6-week-old clean BAlB/c mice were randomly divided into 4 groups:blank group,model group,traditional Chinese medicine group and control group,with 12 mice in each group.After modeling,blank group and model group were given distilled water intragastric.The Chinese medicine group was intragastric administrated with strengthening kidney mixture.The control group was given telmisartan distilled water solution intragastrically.After 8 weeks of intragastric administration,urine,blood and kidney samples were collected at the 9th week,and the pathological changes of glomeruli were observed by he staining.The ratio of urinary albumin to urinary creatinine(ACR)was determined by ElISA.Renal function,including serum creatinine(SCr)and urea nitrogen(BUN),was detected by the kit.The number of TCs was observed by immunofluorescence staining.Ultrastructural changes of renal interstitial TCs were observed by electron microscopy.The protein expression levels of CD34,CD117 and ColIV in renal tissues were detected by Western blotting(WB)before and after treatment.。Results:1.Modeling results of IgAN mice:After 8 weeks of treatment combined with BSA+lPS+CCl4,samples were collected at the end of the 9th week.IgA deposition was observed in the mesangial region of mice in the model group under immunofluorescence test,and mesangial cells and matrix hyperplasia was observed by he staining.Quantitative results of urine protein showed that the level of urine protein in the model group was signIFicantly higher than that in the control group,P<0.01,indicating successful modeling.2.Renal histological changes:compared with the blank group,he staining showed increased mesangial cell prolIFeration and mesangial matrix,renal tubule dilation and lymphocyte infiltration of dIFferent degrees,and some renal tubule atrophy or glomerular segmental sclerosis.After drug intervention,mesangial cells and stroma hyperplasia decreased,and interstitial lesions were alleviated without obvious inflammatory changes.3.Immunofluorescence changes:compared with blank group,there was a large amount of IgA fluorescence deposition in the mesangial region of the model group,and the number of TCs in the renal interstitial was reduced(P<0.01).After drug intervention,compared with model group,the number of renal interstitial TCs in TCM group and control group was increased,P<0.01;4.Transmission electron microscope(TEM)observation revealed that TCs cell body was smaller,nucleus was larger,and cytoplasm around the nucleus was less.The cytoplasm contained relatively aBUNdant mitochondria,endoplasmic reticulum and other organelles in the cytoplasm.Compared with the blank group,the number of TCs and TPs and the number of cell layers in the model group were reduced,the intracellular vesicles of TCs were more,and the intracellular organelles were less.After drug intervention,compared with model group,the number of TCs and TPs cell layers,extracellular vesicles and organelles increased in TCM group and control group,while the number of intracell μ Lar vesicles decreased.5.Urinary albumin creatinine ratio(ACR):Before modeling,the level of urinary protein was low and had no signIFicant dIFference.At the 9th week of modeling,compared with blank group,urine albumin creatinine ratio in model group,Chinese medicine group and control group was signIFicantly increased(P<0.05).After drug intervention,compared with model group,urinary albumin creatinine ratio in TCM group and control group was decreased,and the decrease was more obvious in TCM group,but it was still higher than normal level in both groups,P<0.01;6.Changes of renal function:The levels of SCr and BUN of each group were lower and had no signIFicant dIFference before modeling.At 9 weeks after the completion of modeling,compared with blank group,SCr and BUN levels in model group,TCM group and control group were increased,P<0.05;After drug intervention,compared with model group,SCr and BUN levels in TCM group and control group were decreased compared with model group,and the decrease in TCM group was more obvious,but the two groups were still higher than the normal level,P<0.01;7.Western blotting showed that the expression levels of CD117 and CD34 increased under drug intervention,while the expression levels of Col4 protein decreased and were negatively correlated with CD117 and CD34.Conclusion:1.The number of TCs in renal interstitium of IgAN mouse model decreased and the morphological and structural damage was observed.2.Strengthening kidney Mixture can improve the structure and function of kidney in IgAN mice,increase the number of renal interstitial TCs,and repair the morphological and structural damage of renal interstitial TCs in IgAN mice.