汉防己甲素联合泼尼松对阿霉素肾纤维化大鼠的保护作用及机制

摘要

Objective To explore protective efficacy and possible mechanism of tetrandrine associated with prednisone for re-nal fibrosis rats caused by adriamycin.Method 42 male Sprague -Dawley rats were randomly divided into normal group,renal fi-brosis model group,renal fibrosis rats treated with prednisone、combined prednisone and tetrandrine of there different doses(5,10, 20 mg/kg·d)、prednisone and tetrandrine.it was defined as group A,B,C,D,E,F,G in sequence,the dose of prednisone in the experiment was 10 mg/kg·d.All the rats except group A were given adriamycin of 4 mg/kg、2 mg/kg by tail intravenous injection at the first day and the eighth day respectively,the group A were given the same dose normal saline.From the 8th weekend,the last five groups of rats were treated with drugs mentioned above for eight weeks,group A and group B were treated with normal sa-line.On the 16th weekend,to collect 24 hours′urine of all rats with metabolism cages,kill all the rats and then subsistence all the blood and kidney tissue.The urine protein excretion of all rats were measured,as well as serum,blood urea nitrogen,total ptotein and triglycerides.Kidney pathological changes were observed by HE dying.VG staining to observe expression of collagen fi-bers.The expression of CTGF and MMP13 were determined by immuohistochemistry and RT -PCR.Results Compared with group A,the 24 hours′urine urinary protein content,serum,blood urea nitrogen,triglycerides and CTGF in other groups were higher,the difference was statistically significant (P <0.05),but the total protein and MMP13 were lower,the difference was statistically sig-nificant (P <0.05);compared with group B,the 24 -hour protein,serum,blood urea nitrogen,triglycerides and CTGF in treat-ment group were all cut down and the level of MMP13 contrary to them,the difference was statistically significant (P <0.05);compared with single drug group,the indexes change more significant in group D and E,the difference was statistically significant (P <0.05).Conclusion The mechanism of glucocorticoid combined with tetrandrine inhibition renal tissue fibrosis may be relat-ed to the CTGF and MMP13,midum and high dose tetrandrine has an obvious effect of anti -fibrosis.%目的：研究汉防己甲素联合泼尼松对阿霉素肾纤维化大鼠的保护作用及机制。方法：雄性 SD 大鼠42只随机分为正常对照 A 组、肾纤维化模型 B 组、泼尼松＋汉防己甲素低剂量 C 组[5 mg／（kg·d）]、泼尼松＋汉防己甲素中剂量 D 组[10 mg／（kg·d）]、泼尼松＋汉防己甲素高剂量 E 组[20 mg／（kg·d）]、汉防己甲素 F 组[10 mg／（kg·d）]，泼尼松 G 组，每组6只，实验中所用泼尼松剂量均为10 mg／（kg·d）。于实验开始第1天、8天给予除 A 组外所有大鼠阿霉素（4 mg／kg、2mg／kg）尾静脉注射，A 组给予等量生理盐水，自首次注射阿霉素第8周末给予 C、D、E、F、G 组相应药物灌胃，16周末收集大鼠24 h 尿液，留取血液和肾组织标本。双缩脲法检测大鼠24 h 尿蛋白，全自动生化分析仪检测大鼠血清总蛋白（TP）、血尿素氮（BUN）、血清肌酐（Scr）、三酰甘油（TG）水平。HE 染色和 VG 染色分别观察各组大鼠肾组织病理和胶原纤维的变化。RT －PCR 法检测大鼠肾组织中结缔组织生长因子（CTGF）mRNA、基质金属蛋白酶13（MMP13） mRNA 的表达；免疫组化法检测肾组织中 CTGF 蛋白、MMP13蛋白的表达。结果：与 B 组大鼠24 h 尿蛋白量、BUN 、Scr 、TG、CTGF 表达高于 A 组，差异有统计学意义（P ＜0.05），而 TP、MMP13的水平低于 A 组，差异有统计学意义（P ＜0.05）；C、D、E、F、G 组24 h 尿蛋白量、BUN 、Scr 、TG 、CTGF 表达水平低于 B 组，差异有统计学意义（P ＜0.05），TP、MMP13的水平高于 B 组，差异有统计学意义（P ＜0.05）；与单一用药组比较，联合用药 D 组和 E 组上述指标变化更显著，差异有统计学意义（P 均＜0.05）。结论：汉防己甲素联合泼尼松对阿霉素肾纤维化大鼠有保护作用，其机制可能是抑制促纤维化因子 CTGF 表达，促进抗纤维化因子 MMP13表达；且这种保护作用具有剂量依赖性，以中、大剂量效果较好。