摘要
目的:探讨刺芒柄花素通过调控SIRT1/PGC-1α通路对脓毒症诱导的急性肾损伤(SAKI)的影响及作用。方法:采用盲肠结扎穿刺法(CLP)制备脓毒症大鼠模型,造模成功后选择60只大鼠分为模型组、刺芒柄花素低剂量组、刺芒柄花素中剂量组、刺芒柄花素高剂量组和高剂量刺芒柄花素+SIRT1特异性阻断剂EX527组,另设假手术组,每组12只。腹腔注射刺芒柄花素或EX527进行干预,1次/d,连续3 d。采用全自动生化分析仪检测血清中Cys C、BUN和Scr含量;HE染色观察肾组织病理学变化;DHE探针法检测肾组织中活性氧(ROS)水平;ELISA检测肾组织中丙二醛(MDA)水平及超氧化物歧化酶(SOD)活性;qRT-PCR检测肾组织线粒体DNA(mtDNA)拷贝数以及肾组织中SIRT1 mRNA、PGC-1αmRNA、NRF-1 mRNA及TFAM mRNA表达水平;Western blotting检测肾组织中SIRT1、PGC-1α、NRF-1及TFAM蛋白表达水平。结果:与假手术组比较,模型组大鼠肾组织损伤严重,血清Cys C、BUN、Scr含量明显升高(P<0.01),肾组织线粒体mtDNA拷贝数明显降低(P<0.01),ROS水平、MDA含量明显升高(P<0.01),SOD活性明显降低(P<0.01),同时肾组织中SIRT1、PGC-1α、NRF-1和TFAM mRNA及蛋白表达水平也明显降低(P<0.01)。与模型组比较,刺芒柄花素中、高剂量组大鼠肾损伤明显改善,血清Cys C、BUN、Scr含量均明显降低(P<0.05),肾组织线粒体mtDNA拷贝数明显增加(P<0.05),ROS水平、MDA含量明显降低(P<0.05),SOD活性明显增加(P<0.05),同时肾组织中SIRT1、PGC-1α、NRF-1和TFAM mRNA及蛋白表达水平也明显升高(P<0.05),刺芒柄花素低剂量组大鼠以上各指标与模型组比较,差异均无统计学意义(P>0.05)。然而,EX527干预可明显抑制高剂量刺芒柄花素对SAKI的改善作用。结论:刺芒柄花素可提高肾脏组织线粒体生物合成,减少线粒体氧化应激损伤,进而改善SAKI,其作用机制可能与激活SIRT1/PGC-1α信号通路有关。
Objective: To investigate the effects of formononetin on septic acute kidney injury(SAKI) in rats by regulating SIRT1/PGC-1α pathway and its mechanism. Methods: The rat model of sepsis was established by cecal ligation and puncture(CLP) method. Successful modeling of rats were divided into model group, low dose of formononetin group, medium dose of formononetin group, high dose of formononetin group and high dose of formononetin + SIRT1 specific blocker EX527 group, another sham group was set, with 12 rats in each group.Foroneonetin or EX527 was intraperitoneally injected, once a day for 3 consecutive days. The contents of cystatin C(Cys C), urea nitrogen(BUN) and creatinine(SCR) in serum of rats were detected by automatic biochemical analyzer. Histopathological changes of the renal tissue were observed by HE staining. The level of reactive oxygen species(ROS) in renal tissue of rats was detected by DHE probe. The malondialdehyde(MDA)content and superoxide dismutase(SOD) activity in renal tissues of rats were detected by ELISA. Th e copy number of mitochondrial DNA(mtDNA) and the mRNA expression levels of SIRT1, PGC-1α, NRF-1 and TFAM in renal tissues of rats were detected by qRT-PCR. The protein expression levels of SIRT1, PGC-1α, NRF-1 and TFAM in renal tissues of rats were assessed by Western blotting. Results: Compared with the sham operation group, the renal tissue of the model group was seriously damaged, the contents of serum Cys C, BUN and Scr increased significantly(P<0.01), the copy number of mitochondrial mtDNA in renal tissue decreased significantly(P<0.01), the contents of ROS and MDA increased significantly(P<0.01), and the activity of SOD decreased significantly(P <0.01) The m RNA and protein expression levels of NRF-1 and TFAM were also significantly decreased(P<0.01). At the same time, the m RNA and protein expression levels of SIRT1, PGC-1α,NRF-1 and TFAM in renal tissue were also significantly decreased(P<0.01). Compared with the model group,the renal injury of rats in the medium and high d
作者
周海银
罗兰
陈艳瑛
刘萍萍
肖政辉
方思思
张树菊
ZHOU Hai-yin;LUO Lan;CHEN Yan-ying;LIU Ping-ping;XIAO Zheng-hui;FANG Si-si;ZHANG Shu-ju(Hunan Children's Hospital,Changsha Hunan 410007,China;不详)
出处
《中医药导报》
2022年第1期6-11,共6页
Guiding Journal of Traditional Chinese Medicine and Pharmacy
基金
湖南省中医药科研计划项目(2021036)
湖南省卫生健康委2020年度科研立项课题(20200611)
湖南省科技厅2018年度湖南省重点实验室、工程技术研究中心和科技基础条件平台组建立项(2018TP1028)
2018年度湖南省重点研发计划项目(2018SK2135)。
