The Study Of The Traditional Chinese Medicine Cicada Flower On The Sirt1 Signaling Pathway And Its Apoptosis Mechanism Of Rat Renal Tubular Epithelial Cells (NRK)

Renal interstitial fibrosis(RIF)is the ultimate way and pathological basis for varieties of primary and secondary development of kidney disease leading to end-stage renal disease(ESRD).The degree of renal interstitial fibrosis determines how quickly the development of chronic kidney disease(CKD)and prognosis.Under pathological condition,senescence-associated gene SIRT1 relates to renal tubular epithelial cell,apoptosis,glomeruloscl-erosis and RIF.In recent years,a growing number of studies have shown that in vivo and in vitro Sirtl have clear renal protective effect of Sirtl by regulating kidney cells with diff-erent the stress of the original(oxidative stress,genetic toxicity,and oxygen)reaction to improve the cell survival rate,thus significantly delayed renal interstitial fibrosis process.Compared with cordyceps sinensis,cordyceps cicadae has similar active constituent,but difference flavor and nature.Its sweet flavor and cold-natured characteristic make it fitter to be used under the condition of Yin Deficiency Fire Hyperactivity and arthralgia spasm of kidney collaterals.Research result suggested cordyceps cicadae not only has anti-aging effect,but also can ameliorate glomerulosclerosis and RIF.This topic proposed cicada through regulating Sirtl and downstream pathway,inhibiting renal inherent cells apoptosis and differentiation,play a role of the delay of renal interstitial fibrosis and"hypothesis.In rat renal tubular epithelial cells as the research object,from vitro experiments to explore cicada delay the action mechanism of renal interstitial fibrosis.Objective:To study the cicada hyphae of Ang Ⅱ under the action of renal tubular epithelial cells(NRK)Sirtl protein and its downstream molecules and m-RNA expression of rat renal tubular epithelial cells apoptosis rate and its differentiation degree,reveal the cicada hyphae delay the action mechanism of renal interstitial fibrosis.Methods:We acted on the Angiotensin Ⅱ stimulation of cultured rat renal tubular epithelial cells(NRK)in vitro after 48 h with drug-containing serum,immunoblot and RT-PCR were used to measure the expression of Sirtl and the following protein and mRNA such as smad3,smad7 and p53.TUNEL staining and DAPI redyeing were used to detect apoptosis.We processed with SPSS20.0 statistical software processing,differences between groups using single factor analysis of variance for statistical analysis.Results:①Groups of NRK SIRT1 and m-RNA expression and its downstream proteins:compared with blank group,Ang NRK Ⅱ groups of SIRT1,smad7 protein and m-RNA expression significantly lower(p<0.01);Smad3 and p53 protein and the m-RNA expression significantly raised(p<0.01);Compared with Ang Ⅱ controls,cicada different dose groups,chlorine mitotane group,resveratrol can raise NRK Sirtl,smad7 protein and m-RNA expression(p<0.01),lower NRK of smad3 and p53 protein and the m-RNA expression(p<0.01).The cicada high dose group of medicine is better than chlorine sand mitotane group(p<0.01);Cicada concentration-response relationship between different dose groups.②NRK apoptosis:with the blank control group,Ang Ⅱ group NRK apoptosis rate increased significantly(p<0.01),and Ang Ⅱcontrol group comparison,cicada every dose group could significantly reduce the apoptosis rate of NRK(p<0.01),different doses of the concentration-response relationship between cicada group.Conclusion:This study confirmed that Chinese medicine cicada hyphae can raise the expression of Sirtl,smad7 protein and m-RNA and cut the expression of smad3 and p53 protein and m-RNA,which also have an effection of inhibition of NRK apoptosis.The above experimental results suggest Chinese traditional medicine cicada can play the role of renal tubule interstitial fibrosis to some extent.