汉防己甲素对免疫球蛋白A肾病大鼠肾功能紊乱和肾组织损伤的修复作用  

Repair Effect of Tetrandrine on Renal Dysfunction and Renal Tissue Damage in Rats with Immunoglobulin A Nephropathy

作　　者：马春燕 常婷婷 鲁雅妮 何亚莉 刘曙民 MA Chun-Yan;CHANG Ting-Ting;LU Ya-Ni;HE Ya-Li;LIU Shu-Min(Dept,of Pharmacy,Baoji Central Hospital,Baoji 721000 Shaanxi,China;Dept,of Invasive Technology,Baoji People's Hospital,Baoji 721000 Shaanxi,China)

机构地区：[1]宝鸡市中心医院药剂科,陕西宝鸡721000 [2]宝鸡市人民医院介入科,陕西宝鸡721000

出　　处：《广州中医药大学学报》2021年第8期1666-1672,共7页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：陕西省重点研发计划项目(编号:2017SF-097)。

摘　　要：【目的】探讨汉防己甲素对免疫球蛋白A(IgA)肾病大鼠肾功能紊乱和肾组织损伤的影响。【方法】将60只雄性SD大鼠随机分为正常组,模型组,汉防己甲素5、15、30 mg·kg-1组,每组10只。除正常组,其余各组大鼠均采用脂多糖+牛血清蛋白+四氯化碳法建立IgA肾病模型。成功造模后,汉防己甲素5、15、30 mg·kg-1组分别给予对应剂量的药物腹腔注射,正常组和模型组给予等体积的生理盐水腹腔注射,连续28 d。给药结束后,检测24 h尿蛋白,血清肌酐、尿素氮水平,苏木素-伊红(HE)染色法观察肾组织病理损伤,末端脱氧核苷酸转移酶介导的d UTP缺口末端标记(TUNEL)染色法观察肾组织细胞凋亡情况,酶联免疫吸附分析(ELISA)法检测肾组织和血清中白细胞介素6(IL-6)、白细胞介素10(IL-10)水平,采用水溶性四唑盐1(WST-1)法测定血清总超氧化物歧化酶(SOD)活力,微板法测定乳酸脱氢酶(LDH)活力,硫代巴比妥酸(TBA)法测定丙二醛(MDA)含量,蛋白免疫印迹法检测肾组织Caspase-3、Caspase-9、沉默信息调节因子2相关酶1(SIRT1)、P65、pP65蛋白表达水平。【结果】与正常组比较,模型组大鼠尿蛋白,血清肌酐、尿素氮水平升高,肾组织TUNEL阳性细胞明显增多,肾组织cleaved Caspase-3/Caspase-3、cleaved Caspase-9/Caspase-9比值,肾组织和血清中IL-6水平,血清MDA、LDH水平,肾组织p-P65/P65比值显著升高(均P<0.05),血清IL-10水平亦升高,但差异无统计学意义(P> 0.05),而血清SOD水平、肾组织SIRT1蛋白水平显著降低(均P<0.05)。与模型组比较,汉防己甲素15、30 mg·kg-1组蛋白尿,血清肌酐、尿素氮水平降低,肾组织TUNEL阳性细胞明显减少,肾组织cleaved Caspase-3/Caspase-3、cleaved Caspase-9/Caspase-9比值,肾组织和血清中IL-6水平,血清MDA、LDH水平,肾组织p-P65/P65比值显著降低(均P<0.05),肾组织和血清中IL-10水平,血清SOD水平、肾组织SIRT1蛋白水平显著升高(均P<0.05)。【�Objective To explore the effects of tetrandrine on renal dysfunction and renal tissue damage in rats with immunoglobulin A(IgA)nephropathy. Methods Sixty male SD rats were randomly divided into normal group,model group and tetrandrine 5,15,30 mg·kg-1 groups,10 rats in each group. Apart from the normal group,the rats in the other groups were induced into IgA nephropathy model by lipopolysaccharide plus bovine serum albumin plus carbon tetrachloride method. After successful modeling,tetrandrine 5,15,30 mg·kg-1 groups were given intraperitoneal injection of tetrandrine at corresponding dosage,respectively,and the normal group and model group were given intraperitoneal injection of the same volume of normal saline,the treatment lasting 28 continuous days. After medication, 24-hour urine protein, serum creatinine and urea nitrogen levels were determined,pathological features of renal tissue were observed by hematoxylin-eosin(HE)staining,and apoptosis of renal cells was observed by terminal deoxynucleotidyl transferase(TdT)-mediated dUTP-biotin nick end labeling(TUNEL)assay, levels of interleukin 6(IL-6)and interleukin 10(IL-10)in renal tissue and serum were detected by enzyme-linked immunosorbent assay(ELISA),activity of superoxide dismutase(SOD)in serum was determined by water soluble tetrazolium salt 1(WST-1)assay, activity of lactate dehydrogenase(LDH)was determined by microplate method, content of malondialdehyde(MDA) was determined by thiobarbituric acid(TBA)method,protein expression levels of Caspase-3,Caspase-9,silencing information regulatory protein1(SIRT1),P65 and phosphorylated P65(p-P65)in renal tissue were detected by Western blotting assay. Results Compared with the normal group,the levels of urine protein,serum creatinine and urea nitrogen,and TUNEL positive cells in kidney tissue were significantly increased in the model group,and the ratios of cleaved Caspase-3/Caspase-3 and cleaved Caspase-9/Caspase-9 in kidney tissue,level of IL-6 in kidney tissue and serum,serum MDA and LDH levels,and ratio of p-

关 键 词：汉防己甲素 IgA肾病 肾损伤 肾功能 大鼠 

分 类 号：R285.5[医药卫生—中药学]