Study On The Mechanism Of Chai-ling-tang Effective Parts Group On The Abnormal Proliferation Of Glomerular Mesangial Cells In Rats

Objective:To investigate the effects and mechanism of Chai-ling-tang effective parts group including Chai-Ling small formula(CHLSF) and saikoside-d(SSd) on the abnormal proliferation of glomerular mesangial cells(GMCs) in rats.METHODS:GMCs abnormal proliferation were induced by lipopolysaccharide(1ps).The influence of Chai-ling-tang effective parts group on the abnormal proliferation of GMCs was determined by MTT assay and the expression of the proliferative cell nuclear antigen(PCNA) was determined by immunocytochemical detection;LDH release experiment was used to examine the cytotoxicity of the Chai-ling-tang effective parts group.Reverse transcription polymerase chain reaction(RT-PCR) was used to examine the Cyclin-Kinase Inhibitor(CKI) p21 mRNA at 3h,6h,12h, 24h and 48h after treated by Chai-Ling small formula.Ⅳtype collagen and fibronectin(FN) level in the supernatant of GMCs were determined by enzyme linked immunosorbent assay(ELISA).The activity of Matrix metalloproteinases MMP-2 and MMP-9 were examined by gelatin zymography.RESULTS:thai-ling-tang effective parts group inhibited the proliferation of GMCs in a concentration-dependent manner without obvious cytotoxicity(P＜0.01).Immunocytochemical study for PCNA showed the number of positive cells was significantly decreased in treated group compared with those in control group(P＜0.01). Furthermore,Chai-Ling small formula can significantly stimulated the expression of P21 mRNA determined by RT-PCR(P＜0.05).On the other hand,Chai-ling-tang effective parts group decrease the content of typeⅣcollagen and FN in the supernatants determined by ELISA(P＜0.01) and increased the secretion of MMP-2 and MMP-9.CONCLUSION:thai-Ling small formula could exert an inhibitory effect on the abnormal proliferation of GMCs in a concentration-dependend manner and decrease theⅣtype collagen and FN by increasing the activity of MMP-2 and MMP-9,which strongly suggested its potential therapeutic role in the chronic renal scarring.