Experimental Study On Anti-Renal Fibrosis Effect Of Fresh Clinacanthus Nutans

ObjectiveBased on observing the TGF-?₁-induced HK-2 cell injury model in vitro,combined with the therapeutic effect of fresh Clinacanthus nutans on adriamycin-induced nephrotic syndrome rat model and its corresponding mechanism of action,the fresh Clinacanthus nutans was evaluated The protective effect of renal fibrosis provides experimental basis for the clinical application and further development of Clinacanthus nutans.MethodsThe research mainly divided into two major parts:?1?Experimental study on the inhibitory effect of fresh Clinacanthus nutans juice on HK-2 cells.a Clinacanthus nutans fresh extract of rats fed continuous gavage 7 days,1 h after the last administration of blood serum preparation.b Logarithmic growth phase HK-2 cells group:cell control group,model control group,positive control group,Clinacanthus nutans flower freshjuice?CN?different dose groups,and CN serum high,medium and low dose group.In addition to the normal control group,the other 5 groups were established with TGF-?₁,and the model was terminated 48 hours later.All groups were terminated 48 hours after the corresponding drug intervention.The cell viability and growth status were detected by CCK-8.?2?Pharmacodynamics and Mechanism of the Protection Effect of(CN Solution on Nephrotic Syndrome.The rats were randomly divided into groups according to body weight,ie,blank control group,model control group,prednisone acetate group,CN high dose group,CN medium dose group,and CN low dose group,10 in each group.Except for the blank control group,rat models of nephrotic syndrome were established by tail vein injection of doxorubicin in the other groups,and drug intervention was started at the 5th week after model establishment.After1,2,3,4 and 5 weeks of administration,the rats'body weight and 24-hour urinary protein were measured,and serum albumin,urea nitrogen,creatinine,total cholesterol,and glycerol were measured 5 weeks after administration.Indicators of related effects such as ester,sodium,potassium,chlorine,and kidney index;kidney tissue specimens were subsequently taken for light microscopy examination and immunohistochemistry,and Western blotting was used to observe?-SMA,E-cadherin in kidney tissues.Changes in the expression of cadherin protein content.Results?1?Have significant inhibition of proliferation of HK-2 cells was found in the direct administration of drug-containing serum of Clinacanthus nutans extract and fresh extract of Clinacanthus nutans.?2?The content of microalbuminuria in model group increased gradually,the serum cholesterol,creatinine and urea nitrogen increased,all reached the peak on the 28th day;the content of serum albumin decreased and reached the lowest value on the 28th day.?3?After administration,the amount of urinary albumin in model group increased gradually at 24 hours and peaked on the 4 weeks.In the prednisone group at the same time point,the level of urinary albumin in the model group was significantly lower than that in the model group.Compared with the model group,urinary albumin,serum albumin,cholesterol,urea nitrogen,creatinine were statistically significant.Light microscope showed obvious damage in the model group,and prednisone group,Clinacanthus nutans group,renal injury has been improved.Compared with the model group,the expression of?-SMA protein in the Crocodilus treatment group was significantly lower,and the Clinacanthus nutans high-dose group was more effective and the E-cadherin protein expression was significantly increased.Conclusion?1?Adopting the method of intravenous injection of doxorubicin in two times can successfully replicate the obvious model of adriamycin nephrotic syndrome,and its use time is shorter,which is closer to that of human nephrotic syndrome.?2?Clinacanthus nutans can improve the nephrotic syndrome,high proteinuria symptoms,reduce renal damage,delay nephrotic syndrome renal tubular epithelial cells of renal interstitial fibrosis progression.?3?Clinacanthus nutans can delay renal interstitial fibrosis by up-regulating the expression of E-cadherin protein in renal tissue and decreasing the level of?-SMA protein in renal tubular epithelial cells of rats with nephrotic syndrome.