The Effect Of Panax Notoginseng Saponins On The Synthesis Of Extracellular Matrix In Rats Mesangial Cell Induced By Oxidized Low Density Lipoprotein

Objective : To investigate the effect of different concentration of Panax Notoginseng Saponins on synthesis of extracellular matrix in rats mesangial cell induced by Oxidized Low Density Lipoprotein in different intervention time point,hoping to obtain the laboratory basis of Panax Notoginseng Saponins on the prevention and cure of the glomerular sclerosis,the delay of kidney disease progress and the changes of age-related kidney disease,and to explore a new method for the prevention and cure of kidney disease clinically.Methods:The rats glomerular mesangial cells were cultured in vitro and the subjects were divided into five groups:the control group,the 50 ug/ml Ox-LDL induced group,the 200ug/ml PNS intervention group,the 400ug/ml PNS intervention group,the 800ug/mlPNS intervention group.Each concertration set 3 wells, and each group was detected with 3 time points(24,36,48h).The gene expression of IV collagen,FN,MMP-2,TIMP-2 were detected by the way of RT-PCR;the protein level of IV collagen,FN were detected by the way of ELISA;the protein level of MMP-2,TIMP-2 were detected by the way of Western blotting.Result:The PNS with concertration of 200～800ug/ml acted on the rats glomerular MC which were induced by 50 ug/ml Ox-LDL:(1)the RT-PCR resluts showed that the gene expression of IV collagen,FN,MMP-2,TIMP-2 were significantly different between the induced groups and the controll group in different time point(P<0.05); there were no significant difference between the intervention groups with different concertration and the induced group by the intervention time of 24h(P>0.05);the gene expression of IV collagen,FN,TIMP-2 in PNS intervention group with concertration of 400ug/ml,800ug/ml were lower than the induced group(P<0.05),but the gene expression of MMP-2 were higher than the induced group(P<0.05)by the intervention time of 36h,48h when the cell were collected separately,while the PNS intervention group with concertration of 800ug/ml changed it significantly by the time of 48h;there were no significant difference comparing the PNS intervention group with concertration of 200ug/ml and the induced group in different time points(P>0.05). (2) The cell culture supernatant fluid were collected separately in 24h,36h,48h.The level of IV collagen,FN in supernatant fluid increased along with prolongation of the culture time, presenting time dependency.The level of IV collagen,FN,TIMP-2 in the PNS intervention groups with concertration of 400ug/ml,800ug/ml in 36h were lower than the induced group(P<0.05);the level of IV collagen,FN in the intervention groups with different concertration decreased in 48h compared with the induced group,which the the PNS intervention group with concertration of 800ug/ml decreased the most obviously. (3)the Western Blotting result showed that,the level of TIMP-2 protein in the PNS intervention group with concertration of 400 ug/ml,800ug/ml were lower than the induced group when the PNS intervened 36h,48h separately,but the level of MMP-2 protein were higher than the induced group(P<0.05),while the PNS intervention group with concertration of 800ug/ml changed it more significantly。There is no significant difference between the PNS intervention group with concertration of 200ug/ml with the induced group in different time points.Conclusion:(1) The mRNA expression and the secretion of protein of IV collagen,FN,TIMP-2 in the rats glomerular mesangial cells increased obviously which induced by OX-LDL with concertration of 50ug/ml,but the mRNA expression and the secretion of MMP-2 decreased obviously.(2) Panax Notoginseng Saponins could suppress obviously the synthesis of extracellular matrix in rats mesangial cell induced by Oxidized Low Density Lipoprotein,presenting time dependence,which the intervention group with concertration of 800ug/ml was the most obvious.(3) Panax Notoginseng Saponins could spur obviously the synthesis of MMP-2 in the rats kidney mesangial cell induced by Oxidized Low Density Lipoprotein,and suppress obviously the TIMP-2,which both of them presented time-concertration dependency,which the intervention group with concertration of 800ug/ml was the most obvious.