三七总皂苷对顺铂致肾损伤大鼠肾组织纤维化的改善作用及对相关因子表达的影响

目的:研究三七总皂苷(PNS)对顺铂致肾损伤模型大鼠肾组织纤维化的改善作用及对相关因子的影响。方法:将72只SD大鼠随机分为空白组、模型组、阳性药物组和PNS低、中、高剂量组,每组12只。除空白组外,其余各组大鼠采用顺铂尾静脉注射(3 mg/kg×4次)建立肾损伤模型。从首次注射顺铂后第1天开始,阳性组大鼠腹腔注射安磷汀溶液(1.0 mg/kg),PNS各剂量组大鼠腹腔注射PNS溶液(15.63、31.35、62.70 mg/kg),空白组和模型组大鼠注射等体积生理盐水,给药体积均为0.2 mL,连续给药60 d。收集大鼠24 h尿液,检测β-N-乙酰胺基葡萄糖苷酶(NAG)、24 h尿蛋白(Upro/24 h)含量;检测大鼠血清中肌酐(Scr)、尿素氮(BUN)含量;采用逆转录荧光聚合酶链式反应(RT-PCR)和免疫组化法分别检测大鼠肾组织中结缔组织生长因子(CTGF)、转化生长因子β_1(TGF-β_1)、Ⅰ型胶原(Col-1)、金属蛋白酶组织抑制因子1(TIMP-1)、纤溶酶原激活物抑制物1(PAI-1)的mRNA和蛋白表达水平。结果:与空白组比较,模型组大鼠尿液中NAG、Upro/24 h含量,血清中Scr、BUN含量,以及肾组织中CTGF、TGF-β_1、Col-1、TIMP-1、PAI-1的mRNA和蛋白表达水平均显著升高(P<0.05)。与模型组比较,PNS各剂量组大鼠上述尿液和血清生化指标含量均显著降低;PNS各剂量组大鼠肾组织中CTGF、TGF-β_1的mRNA表达水平和CTGF、TGF-β_1、Col-1、TIMP-1蛋白表达水平,PNS高剂量组Col-1的m RNA表达水平,PNS中、高剂量组TIMP-1的mRNA表达水平和PAI-1蛋白表达水平均显著降低(P<0.05)。与阳性组比较,PNS中、高剂量组大鼠尿液中NAG、Upro/24 h含量显著降低(P<0.05)。结论:PNS能有效改善顺铂致肾损伤模型大鼠的肾功能,其可能通过下调肾组织中肾纤维化相关性因子CTGF、TGF-β_1、Col-1、TIMP-1、PAI-1的表达,从而发挥减轻顺铂致肾纤维化的作用。

Improvement Effects of Panax Notoginsenosides on Renal Fibrosis in Cisplatin-induced Renal Injury Rats and Its Effects on the Expression of Renal Fibrosis Related Factors

OBJECTIVE:To study improvement effects of Panax notoginsenoside(PNS) on cisplatin-induced renal injury model rats and its effects on related factors. METHODS:Totally 72 SD rats were randomly divided into blank group,model group,positive drug group and PNS low-dose,medium-dose,high-dose groups,with 12 rats in each group. Except for blank group,other groups were given cisplatin via tail vein(3 mg/kg×4 times)to establish renal injury model. Since the first day after the first injection of cisplatin,positive group was given anfostine solution intraperitoneally(1.0 mg/kg);PNS groups were given PNS solution intraperitoneally(15.63,31.35,62.70 mg/kg);blank group and model groups were given constant volume of normal saline 0.2 mL,for consecutive 60 d. The 24 h urine of rats was collected;the contents of β-N-acetylaminoglycosidase(NAG)and 24 h urine protein(Upro/24 h)were detected;the serum contents of Scr and BUN were detected. mRNA and protein expression of CTGF,TGF-β 1,Col-1,TIMP-1 and PAI-1 in renal tissue were determined by RT-PCR and immunohistochemistry respectively. RESULTS:Compared with blank group,the contents of NAG and Upro/24 h in urine,serum contents of Scr and BUN,mRNA and protein expression levels of CTGF,TGF-β1,Col-1,TIMP-1 and PAI-1 in renal tissue were increased significantly(P<0.05). Compared with model group,the contents of above urine and serum biochemical indicators were decreased significantly in PNS groups;mRNA expression of CTGF and TGF-β1 and protein expression of CTGF,TGF-β1,Col-1 and TIMP-1 in renal tissue of rats in PNS groups,mRNA expression of Col-1 in PNS high-dose group,and mRNA expression of TIMP-1 and protein expression of PAI-1 in PNS medium-dose and high-dose groups were decreased significantly(P<0.05). Compared with positive group,the contents of NAG and Upro/24 h in urine were decreased significantly in PNS medium-dose and high-dose groups(P< 0.05). CONCLUSIONS: PNS can effectively improve the renal function of cisplatin-induced renal injury model rats,and relieve cisplatin-induced renal fibrosis by decreasing the expression of renal fibrosis related factors as CTGF,TGF-β1,Col-1,TIMP-1 and PAI-1 in renal tissue.