A Single Clinical Dose Of Cisplatin Induced Renal Interstitial Fibrosis In Rats And Improvement Effect Of Ginaton Injection

Objectives:1.To investigate the effect of a single clinical dose of cisplatin on renal interstitial fibrosis in rats.2.To investigate the effect of Ginaton injection on cisplatin-induced renal interstitial fibrosis in rats and its possible mechanism,providing a theoretical basis for the clinical treatment of cisplatin-induced renal interstitial fibrosis.Methods:1.Renal interstitial fibrosis in rats induced by a single clinical dose of cisplatin:96 male SD rats were randomly divided into control group and cisplatin group,with 48 rats in each group.Control group and cisplatin group were given equal volume of normal saline and cisplatin 5 mg/kg intraperitoneally on day 1,respectively.At days 8,14,21,30,40,50,60 and 90,six rats of each group were sacrificed.The levels of BUN and Scr were measured by automatic biochemical analyzer.HE staining was used to assessing the degree of tubulointerstitial damage.Masson’s trichrome staining was employed to evaluate the relative area of tubulointerstitial fibrosis.Immunohistochemical staining was used to detect the expression of α-SMA,Col I and TGF-β1 in renal tissues.2.Ginaton injection improved cisplatin-induced renal interstitial fibrosis in rats:45 male SD rats were randomly divided into control group,cisplatin group,cisplatin + L-EGb group,cisplatin + M-EGb group and cisplatin + H-EGb group,with 9 rats in each group.Control group was given equal volume of normal saline intraperitoneally on day 1 and days 22 to 40,once a day.Cisplatin group was given cisplatin 5 mg/kg intraperitoneally on day 1 and equal volume of normal saline from 22 to 40 days,once a day.Cisplatin + L-EGb group(1.58mg/kg),cisplatin + M-EGb group(3.17mg/kg)and cisplatin + H-EGb group(6.34mg/kg)were received cisplatin 5 mg/kg intraperitoneally on day 1 and EGb from 22 to 40 days,once a day.All rats were sacrificed on day 40.On day 40,all rats were sacrificed to determined the levels of BUN and Scr,and the urinary level of NAG.HE staining was used to assessing the degree of tubulointerstitial damage.Masson’s trichrome staining was employed to evaluate the relative area of tubulointerstitial fibrosis.Immunohistochemical staining was used to detect the expression of α-SMA and Col I in renal tissues.3.The mechanism of the effect of Ginaton injection on cisplatin-induced renal interstitial fibrosis in rats:Immunohistochemical staining was used to detect the expression of TGF-β1 in renal tissues.Western blot was used to detect the protein expression of TGF-β1,p38 MAPK and p-p38 MAPK.The mRNA expression of p38 MAPK was detected by qRT-PCR.Results:1.Renal interstitial fibrosis in rats induced by a single clinical dose of cisplatin:Compared with the control group,the levels of BUN and Scr,tubulointerstitial injury index,relative area of tubulointerstitial fibrosis,and the expression of α-SMA,Col I and TGF-β1of rats in the CDDP treated group at each time point were significantly increased(P<0.05 or P<0.01).2.Ginaton injection improved cisplatin-induced renal interstitial fibrosis in rats:Compared with the control group,the levels of BUN and Scr,the urinary level of NAG,tubulointerstitial injury index,relative area of tubulointerstitial fibrosis,and the expression of α-SMA and Col I of rats in the cisplatin group were significantly increased(P<0.01).Compared with the cisplatin group,the levels of BUN and Scr,the urinary level of NAG,tubulointerstitial injury index,relative area of tubulointerstitial fibrosis,and the expression of α-SMA and Col I of rats in the cisplatin + L-EGb group,cisplatin + M-EGb group and cisplatin + H-EGb group were significantly decreased(P<0.05 or P<0.01).3.The mechanism of the effect of Ginaton injection on cisplatin-induced renal interstitial fibrosis in rats:Compared with the control group,the expression of TGF-β1,the ratio of p-p38 MAPK to p38 MAPK and the mRNA expression of p38 MAPK of rats in the cisplatin group were significantly increased(P<0.01).Compared with the cisplatin group,the expression of TGF-β1,the ratio of p-p38 MAPK to p38 MAPK and the mRNA expression of p38 MAPK of rats in the cisplatin + L-EGb group,cisplatin + M-EGb group and cisplatin + H-EGb group were significantly decreased(P<0.05 or P<0.01).Conclusions:1.A single clinical dose of cisplatin can induce renal interstitial fibrosis in rats.2.Ginaton injection can significantly improve the cisplatin-induced renal interstitial fibrosis in rats.3.The mechanism of the effect of Ginaton injection on cisplatin-induced renal interstitial fibrosis in rats may be associated with the inhibition of TGF-β 1/p38 MAPK pathway.