滋阴清热复方、NAC与mitoQ对地塞米松所致MRL/lpr小鼠肾脏线粒体功能异常影响的研究  

Effects of Compound Ziyin Qingre Prescription, NAC and mitoQ on abnormal renal mitochondrial function in dexamethasone-induced MRL/lpr mice

作　　者：赵晓庆 党若楠 程喜平[2] 宋兴华[2] 黄杏兰 黄彩凤 刘升 王杏容 颜鹏 杨裕祺 范萍[2] 周毅业 张穗盈 Zhao Xiaoqing;Dang Ruonan;Cheng Xiping;Song Xinhua;Huang Xing Lan;Huang Caifeng;Liu Sheng;Wang Xingrong;Yang Yuqi;Fan Ping;Zhou Yiye;Zhang Suiying(The Affiliated TCM Hospiotal of Guangzhou Medical university,Guangzhou 510180,China;The First Affiliated Hospital of Guangzhou Medical University,Guangzhou 510120,China;Guangzhou Twelfth People's Hospital,Guangzhou 510515,China;SSL Central Hospital of Dongguan City,Dongguan 523127,China)

机构地区：[1]广州医科大学附属中医医院,广州510180 [2]广州医科大学附属第一医院,广州510120 [3]广州市第十二人民医院,广州510515 [4]广东省东莞市松山湖中心医院,广东东莞523127

出　　处：《国际免疫学杂志》2022年第6期564-574,共11页International Journal of Immunology

基　　金：国家自然科学基金(81373649,81673983,82074172);广东省钟南山医学基金会(07009)。

摘　　要：目的探讨滋阴清热复方、N-乙酰半胱氨酸(N-Acetyl-L-cysteine,NAC)与米托蒽醌甲磺酸盐(mitoquinone mesylate,mitoQ)对地塞米松所致MRL/lpr小鼠线粒体功能异常影响。方法50只8周龄的MRL/lpr小鼠随机分为狼疮鼠组、地塞米松组、滋阴清热复方+地塞米松组、NAC+地塞米松组、mitoQ+地塞米松组,每组10只;另选Balb/c小鼠为空白对照组,共10只;分别于干预12周后,取肾脏组织,通过实时荧光定量聚合酶链反应(real-time quantitative polymerase chain reaction,qRT-PCR)与蛋白质印迹(Western-blot,WB)检测线粒体基因细胞色素C氧化酶亚基1(MT-CO1),线粒体自噬基因PTEN诱导激酶1(PTEN-induced putative kinase 1,PINK1)、白细胞介素(interleukin,IL)-1β的mRNA及蛋白表达水平;比色法检测丙二醛(malondialdehyde,MDA)、超氧歧化酶(superoxide dismutase,SOD)表达差异。结果与正常鼠组相比,狼疮鼠组肾脏线粒体呼吸链基因MT-CO1表达降低[qRT-PCR:(1.00+0.12)比(0.39±0.05);WB:(1.73±0.14)比(0.75±0.08),t值分别为9.61,10.60,P值均<0.05]、线粒体复合物Ⅳ活性减弱[(9.37±0.32)比(7.12±0.26),t=9.53,P<0.05];SOD活性降低[(20.62±3.97)比(15.04±0.33),t=2.80,P<0.05],MDA含量升高[(21.90±2.39)比(41.51±4.74),t=7.40,P<0.05];线粒体自噬基因PINK1表达升高[qRT-PCR:(0.99±0.17)比(1.42±0.05);WB:(0.80±0.06)比(1.37±0.07),t值分别为4.11,10.57,P值均<0.05],炎症因子IL-1β表达升高[qRT-PCR:(1.04±0.32)比(3.43±0.66);WB:(2.48±0.15)比(3.20±0.24),t值分别为6.48,4.45,P值均<0.05],差异具有统计学意义。与狼疮鼠组相比,地塞米松组肾脏线粒体呼吸链基因MT-CO1表达[qRT-PCR:(0.39±0.05)比(0.19±0.05);WB:(0.75±0.08)比(0.36±0.08),t值分别为5.49,5.79,P值均<0.05]、线粒体复合物Ⅳ活性减弱[(7.12±0.26)比(5.83±0.39),t=5.56,P<0.05],SOD(U/mg)活性降低[(15.04±0.33)比(10.28±1.16),t=7.94,P<0.05]、MDA(μmol/mg)蛋白含量升高[(41.51±4.74)比(65.91±6.22),t=6.24,P<0.05];线粒体自噬基因PINK1表达升高[qObjective Exploring the effects of Compound Ziyin Qingre Prescription,N-Acetyl-L-cysteine(NAC)and Mitoquinone mesylate(mitoQ)on abnormal mitochondrial function in dexamethasone-induced MRL/lpr mice.Methods Fifty 8-week-old MRL/lpr mice were randomly divided into lupus mouse group,dexamethasone group,Compound Ziyin Qingre Prescription+dexamethasone group,NAC+dexamethasone group,and mitoQ+dexamethasone group.Ten mice in each group;Balb/c mice were selected as blank control group,10 mice in total,after 12 weeks of intervention respectively,the kidney tissues were taken and the mRNA and protein expression levels of mitochondrial gene MT-CO1,mitochondrial autophagy gene PINK1,inflammatory factor interleukin(IL)-1βwere detected;colorimetric method was used to detect the difference of malondialdehyde(MDA)and superoxide dismutase(SOD)expression.Results Compared with normal mice,the renal mitochondrial respiratory chain gene MT-CO1 expression[RT-qPCR:(1.00+0.12)vs(0.39±0.05);WB:(1.73±0.14)vs(0.75±0.08),t values were 9.61,10.60,both P values<0.05],mitochondrial complexⅣactivity and SOD activity were decreased[(9.37±0.32)vs(7.12±0.26),(20.62±3.97)vs(15.04±0.33),t values were 9.53,2.80,both P values<0.05],MDA was increased[(21.90±2.39)vs(41.51±4.74),t=7.40,P<0.05],mitochondrial autophagy gene PINK1 expression was increased[RT-qPCR:(0.99±0.17)vs(1.42±0.05);WB:(0.80±0.06)vs(1.37±0.07),t values were 4.11,10.57,both P values<0.05]and inflammatory factor IL-1βexpression was increased[RT-qPCR:(1.04±0.32)vs(3.43±0.66),WB:(2.48±0.15)vs(3.20±0.24),t values were 6.48,4.45,both P values<0.05]in the lupus group.Compared with MRL/Lpr mice,the dexamethasone group showed decreased mitochondrial respiratory chain gene expression MT-CO1[RT-qPCR:(0.39±0.05)vs(0.19±0.05),WB:(0.75±0.08)vs(0.36±0.08),t values were 5.49,5.79,both P values<0.05],mitochondrial complexⅣ[(7.12±0.26)vs(5.83±0.39),t=5.56,P<0.05]activity and SOD activity[(15.04±0.33)vs(10.28±1.16),t=7.94,P<0.05],increased MDA[(41.51±4.74)vs(65.91±6.22),t

关 键 词：系统性红斑狼疮 线粒体功能 滋阴清热复方 

分 类 号：R285.5[医药卫生—中药学]