Study On The Mechanism By Which Qiteng Xiao Zhuo Granule Treats Chronic Glomerulonephritis Based On Syk/Ras/c-Fos Signaling Pathway

Objective Chronic glomerulonephritis(CGN)belongs to the traditional C hinese medicine(TCM)category of “spleen deficiency,damp and blood stasis”.A theory of “invigorating the spleen and water,removing blood stasis” was put forward by a famous elder the traditional C hinese medicine doctor.Q i Teng Xiao Zhuo granules,was originated from this theory,and it has been commonly recognized as a prescription that has curative effects in clinical practice,which proved that it could treat CGN.However,the molecular mechanisms still remain unknown.The aim of this study was to investigate the effect of Q iteng Xiaozhuo granule on expression of m RNA and protein of Syk/Ras/c-Fos signal pathway,and analyze the possible molecular mechanisms of Qi Teng Xiao Zhuo granule mediated treatment of CGN.Method 1 Effect of Q iteng Xiaozhuo granule on the regulatory Syk/Ras/c-Fos signal pathway in C GN rats which were induced by ADR The model of CGN rats was made using tail vein injection of adriamycin(ADR).After model establishment,rats were randomly divided into model group,Q iteng Xiaozhuo granule(21.6?10.8?5.4 g/kg)group and Huangkui capsule(1.0 g/kg)group,once daily for thirty days.Normal rats were taken as normal group.The normal and model groups were given an equivalent solvent.Histological analysis of kidney tissue was by hematoxylin-eosin staining(HE).The 24-hour urinary protein,N-acetyl-?-Dglucosaminidase(NAG),blood urea nitrogen(BUN),serum creatinine(Scr),retinolbinding protein(RBP)and cystatin c(C ys-c)was detected by automatic biochemical.RT-q PCR method was undertaken to determine the expression of Syk,Ras,MEK1/2,ERK1/2,c-Fos m RN A in kidney tissue.Immunohistochemical(IHC)and Western blot(WB)method were undertaken to determine the expression of p-Syk,Ras,p-MEK1/2,p-ERK1/2,c-Fos protein in kidney tissue.2 Effect of medicated serum prepared with Q iteng Xiaozhuo Granule on the regulatory Syk/Ras/C-Fos signal pathway in HBZY-1 which were stimulated by LPS HBZY-1 cell were cutured in DMEM containing in 37 ? incubator and the model of cell was stimulated by LPS.The MTT method was used to screen out the best dosage and stimulation time of Q iteng Xiaozhuo granule medicated serum.HBZY-1 cell were divided into normal group,LPS group,LPS+ normal serum group,LPS+medicated serum group,LPS+R406 group,LPS+ medicated serum+R406 group.After intervention,RT-q PCR method was undertaken to determine the expression of Syk,Ras,MEK1/2,ERK1/2,c-Fos m RN A,and Western blot(WB)method were undertaken to determine the expression of p-Syk,Ras,p-MEK1/2,p-ERK1/2,c-Fos protein.Results 1 Effect of Q iteng Xiaozhuo granule on the regulatory Syk/Ras/c-Fos signal pathway in C GN rats which were induced by ADR There was little effect on the index of renal function in the treatment groups,which indicated that the pathological changes of kidney could be significantly reduced.The expression levels of 24 h urine protein and NAG in urine,BUN,Scr,RBP,Cys-c in serum,Syk,Ras,MEK1/2,ERK1/2,c-Fos m RNA and p-Syk,Ras,p-MEK1/2,p-ERK1/2,c-Fos protein of kidney tissue in model group were higher than that of control group.The above indexes were ameliorated in middle and high dose of Q iteng Xiaozhuo granule groups.2 Effect of medicated serum prepared with Q iteng Xiaozhuo Granule on the regulatory Syk/Ras/c-Fos signal pathway in HBZY-1 which were stimulated by LPS The optimal dosage of Q iteng Xiaozhuo granule medicated serum was 40% and reasonable stimulation time was 48 h.Under these conditions,the cell inhibition rate(IR)was higher.Q iteng Xiaozhuo granule medicated serum reduced significantly the expression of Syk,Ras,MEK1/2,ERK1/2,c-Fos m RN A and p-Syk,Ras,p-MEK1/2,p-ERK1/2,c-Fos protein in HBZY-1.Conclusion 1 Syk/Ras/c-fos signaling pathway has changed significantly in the developing of CGN.2 The regulation of Syk/Ras/c-Fos signaling pathway by Q iteng Xiaozhuo granule may be one of its mechanisms in the treatment of CGN.