The Mechanism Of IgA Nephropathy Is Regulated By The TGF-?1/Smads Signaling Pathway Based On The Study Of "renal Collateral Disease"

Purpose:Observation with"kidney disease"for the principle of formula Qijishenkang decoction of transgenic mice IgAN TGF-?1/Smads signal transduction pathways and Ang?induced human glomerular mesangial cells FN,TGF-?1.To explore the difference of efficacy of Qijishenkang decoction(low,medium and high dose)on IgAN at different time points and its mechanism of TCM efficacy,to verify the scientific hypothesis of"filling,clearing and eliminating"kidney collaterals theory,and to explore the pathological mechanism of IgAN.Material and method:1.Construction and evaluation of transgenic IgAN mice,grouping and detection methods:In vivo experiment:constructed transgenic parent mice containing human IgAN,bred and identified F1 generation genotype-positive Ig HA1-mut mice,normal group,F1 generation mice randomly were divided into model group,western medicine group and Qijishenkang decoction(low,medium and high dose groups),with 8 mice in each group.After intragastric administration,kidney specimens of mice were collected at 14,21 and 28d.Smad7m RNA and Smad3m RNA levels were observed by RT-PCR,and TGF-?1 protein and Smad2 protein expression were measured by Western-blot.2.Preparation of pharmacological serum of Qijishenkang decoction:In vitro experiment:the decoction room was decocted and prepared Qijishenkang traditional Chinese medicine decoction,telmisartan tablets were dissolved in distilled water.Forty SD male rats were randomly divided into western medicine group,normal group,and Qijishenkang decoction group(low,medium and high dose groups).After intragastric administration for a consecutive week,the blood was collected from the abdominal aorta,inactivated in water bath and sterilized,drug-containing serum was prepared and refrigerated at-20?.3.Culture of HMC,grouping and detection methods:In vitro experiment:human glomerular mesangial cells in culture bottle(75 cm²)vaccination and RPMI-1640 culture are added into the culture bottle(containing 10%FBS),at 37?,5%CO₂incubator for culture,Ang?promote human glomerular mesangial cells growth.Pharmacological serum was applied to western medicine group and Qijishenkang decoction group(low,medium and high dose groups).At 24h and 48h,the growth of mesangial cells of human glomerular was observed by MTT method,and the contents of TGF-?1 and FN were detected by ELISA method.Results:1.The results of RT-PCR and Western-blot in vivo showed that On 14,21 and 28 days,compared with normal group,the expression of TGF-?1,Smad2 protein and Smad3m RNA in IgAN mice(model group)increased(P<0.05),and the longer the time was,the higher the expression was,while the Smad7m RNA expression was decreased(P<0.05).the results of Qijishenkang decoction group and the western medicine group were reversed(P<0.05)2.The results of MTT and ELISA in vitro showed that At 24h and 48h,OD value in the model group was higher than the normal group(P<0.05),western medicine group,Qijishenkang decoction group were less than model group(P<0.05).The levels of FN and TGF-?1 in the model group were higher than normal(P<0.01).Qijishenkang decoction in each groups were lower than model group(P<0.01).the medium and high dose groups were lower than the western medicine group(P<0.05).Conclusion:1.To clarify that one of the mechanisms of IgAN action is related to TGF-?1/Smads signaling pathway,FN and other factors.2.It is indicated that Qijishenkang decoction of different concentrations can regulate TGF-?1/Smads signaling pathway,inhibit FN overexpression,reduce glomerular mesangial accumulation,improve the pathological changes of IgAN,and protect the kidney.3.Prove to"Fill?clear?eliminate kidney collaterals"method for guiding theory of Qijishenkang decoction on experimental IgAN transgenic mice and inhibit Ang?induced human glomerular mesangial cells proliferation experiment curative effect,perfecting the theory of"kidney disease".