丹酚酸A、C分子药对配伍干预肾纤维化组织凋亡相关蛋白caspase-3、GRP78的实验研究

Experimental study on compatibility of Salvianolic Acid A and C on pyroptotic proteins caspase-3 and GRP78 in renal fibrosis

目的研究丹酚酸A、C分子药对配伍对肾纤维化过程中凋亡相关蛋白半胱氨酸天门冬氨酸蛋白酶3(caspase-3)、葡萄糖调节蛋白78(GRP78)的调节作用。方法将50只雄性SD大鼠随机分为5组,即正常组,模型组,丹酚酸A组,丹酚酸C组,丹酚酸A、C按1:1配伍组(丹酚酸A+C组)。除正常组外,其余大鼠均采用单侧输尿管梗阻(UUO)造模。造模后第2天开始各组给予相应的干预措施;2周后处死大鼠,收集相应标本,检测尿β2-微球蛋白(β2-MG)、N-乙酰-β-D氨基葡萄糖苷酶(NAG),光镜观察肾组织病理,TUNEL检测技术观察肾小管间质细胞凋亡情况,实时荧光定量PCR(RT-PCR)技术检测肾组织caspase-3、GRP78 mRNA的表达。结果 1与正常组比较,模型组及各治疗组尿β2-MG、NAG水平均明显升高(P<0.05)。2与模型组比较,各治疗组尿β2-MG水平均明显降低(P<0.05),丹酚酸A、C组尿NAG水平明显降低(P<0.05)。3与模型组比较,各治疗组肾脏组织病理情况均有不同程度的改善,尤以丹酚酸A、A+C组改善明显(P<0.05)。4丹酚酸A、C组及A+C组可以下调caspase-3、GRP78的表达,有效减轻肾小管间质细胞凋亡,且丹酚酸A+C组较丹酚酸C组效果更明显(P<0.05)。结论丹酚酸A、C及A+C分子药对配伍可一定程度保护UUO大鼠肾小管功能,并下调肾组织caspase-3、GRP78的表达,减少肾小管间质细胞凋亡的发生,减轻肾间质纤维化程度。上述作用可能与该药物干预凋亡相关蛋白caspase-3、GRP78的表达,抑制内质网应激途径有关。

Objective To investigate the intervention effects of Salvianolic acid A and C on drug compatibility of pyroptotie proteins caspase-3 and GRP＇78 of renal fibrosis in rats. Methods Fifty SD rats were randomly divided into 5 groups ： normal group, model group, Salvianalic acid A group, Salvianolic acid C group, Salvianolic acid A ＋ C group. Except the normal group, the unilateral ureteral obstruction （UUO） rat models were established. The next day after establishment of the model, corresponding interventions were given to each group, and then the rats were killed for samples collection. The urinary β2-Microglobutin （β2-MG） and N-acetyl-β-D-glucosaminidase （NAG） were detected. The nephridial tissues were made into paraffin section by dehydration and embedding which were used for HE staining and TUNEL detection. And the expression of caspase-3 and GRP78 were analyzed by Real-time quantitative RT-PCR. Results （1） Compared with the normal group, the levels of β2-MG and NAG in the model group and treatment groups were markedly increased （ P 〈 0.05 ）. （2） Compared with the model group, β2-MG level in the treatment groups were markedly reduced （ P 〈 0.05 ）, and NAG level in the Saivianolic acid A group and Salvianalic acid C group were reduced significantly （P 〈 0.05 ）. （3） Compared with the model group, pathologic condition of kidney tissues in the treatment groups was improved, especially in the Salvianolic acid A group and A ＋ C group （ P 〈 0.05 ）. （4） Compatibility of Salvianolic acid A, C and A ＋ C can down-regulate the expressions of caspase-3 and GRP78, and alleviate renal tubular cell apoptosis. The A ＋ C group was more significant than that of the C group （ P 〈 0.05 ）. Conclusion Salvianolic acid A and C component molecules of drug compatibility could protect renal tubular function in a certain degree, and suppress the expression of easpase-3 and GRP78, significantly reduce renal tubular cell apoptosis and alleviate renal tubulointerstitial fibrosis. This mechanism might be related to the intervention on the expression of caspase-3 and GRP78, and then endoplasmic reticulum stress pathway was suppressed.