三七总皂苷基于PI3K/Akt/mTOR通路调控自噬和泛素蛋白积累对大鼠肾缺血再灌注损伤的影响  

Effect of panax notoginseng saponins regulating autophagy and ubiquitin protein accumulation on renal ischemia-reperfusion injury in rats via activating the PI3K/Akt/mTOR pathway

作　　者：王华[1] 谭超[1,2,3] 王子焱 张涛 李鑫成 WANG Hua;TAN Chao;WANG Ziyan;ZHANG Tao;LI Xincheng(The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410007,Hunan,China;Inherit Workroom of Medical Master Professor XIONG Jibo’s Experiences,Changsha 410007,Hunan,China;The Domestic First-class Discipline Construction Project of Chinese Medicine of Hunan University of Chinese Medicine,Changsha 410208,Hunan,China;Graduate School,Hunan University of Chinese Medicine,Changsha 410036,Hunan,China;Medical School,Hunan University of Chinese Medicine,Changsha 410036,Hunan,China)

机构地区：[1]湖南中医药大学第一附属医院,湖南长沙410007 [2]国医大师熊继柏教授传承工作室,湖南长沙410007 [3]湖南中医药大学中医学国内一流建设学科,湖南长沙410208 [4]湖南中医药大学研究生院,湖南长沙410036 [5]湖南中医药大学医学院,湖南长沙410036

出　　处：《现代中西医结合杂志》2023年第11期1461-1467,1472,共8页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：湖南省自然科学基金项目(2021JJ70112);湖南省教育厅大学生创新训练项目(020000201326)。

摘　　要：目的基于磷酸肌醇3激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白(mTOR)通路探究三七总皂苷调控自噬和泛素蛋白积累对大鼠肾缺血再灌注损伤的保护作用。方法将30只SPF级SD大鼠随机分为模型组、三七总皂苷低剂量组、三七总皂苷高剂量组,每组10只。3组大鼠均进行肾缺血再灌注造模,三七总皂苷低、高剂量组分别在缺血造模前1 h尾静脉注射三七总皂苷10 mg/kg、20 mg/kg,之后连续3 d尾静脉注射等剂量三七总皂苷。实验结束后,ELISA法检测各组大鼠血清尿素氮(BUN)、肌酐(Cr)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-18(IL-18)水平,苏木精-伊红染色观察肾组织病理形态,试剂盒检测肾组织中丙二醛(MDA)、活性氧(ROS)、超氧化物歧化酶(SOD)、乳酸脱氢酶(LDH)含量,Western blot法检测肾组织中PI3K/Akt/mTOR信号通路和凋亡蛋白表达情况,RT-PCR法检测肾组织中自噬相关蛋白和泛素化蛋白mRNA表达情况。结果三七总皂苷高剂量组和三七总皂苷低剂量组BUN、Cr、IL-1β、IL-6、IL-18水平均明显低于模型组(P均<0.05),病理损伤明显轻于模型组;三七总皂苷高剂量组和三七总皂苷低剂量组肾组织中MDA、ROS、LDH含量和p-mTOR、半胱氨酸天门冬氨酸酶3(Caspase-3)、Bcl-2关联蛋白X(Bax)蛋白相对表达量均明显低于模型组(P均<0.05),SOD含量和p-Akt、PI3K、B细胞淋巴瘤基因-2(Bcl-2)蛋白相对表达量均明显高于模型组(P均<0.05);三七总皂苷高剂量组和三七总皂苷低剂量组肾组织中Beclin-1、自噬相关蛋白8(ATG8)、微管相关蛋白轻链3(LC3)Ⅱ/Ⅰ、肿瘤坏死因子相关因子6(TRAF6)、Beclin-1调节自噬蛋白-1(AMBRA1)mRNA相对表达量均明显高于模型组(P均<0.05),神经前体细胞表达发育下调因子4(NEDD4)mRNA相对表达量均明显低于模型组(P均<0.05)。结论三七总皂苷可通过激活PI3K/Akt/mTOR通路,促进泛素蛋白酶系统的表达并诱导自�Objective It is to explore the protecting effect of panax notoginseng saponins(PNS)regulating autophagy and ubiquitin protein accumulation on renal ischemia-reperfusion injury in rats via activating the phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)pathway.Methods Thirty SPF SD rats were randomly divided into 3 groups:model group,NPS low-dose group,PNS high-dose group,with 10 rats in each group.All the rats of the 3 groups were used to establish renal ischemia-reperfusion models,and the low and high dose groups of PNS were injected with PNS 10 mg/kg and 20 mg/kg into the tail vein 1h before the ischemia modeling,respectively,and then intraperitoneally injected with equal doses of PNS for 3 consecutive days.At the end of the experiment,the levels of serum urea nitrogen(BUN),creatinine(Cr),interleukin-1β(IL-1β),interleukin-6(IL-6)and interleukin-18(IL-18)in the rats of each group were detected by ELISA,the pathological morphology of renal tissues was observed by hematoxylin-eosin staining,and the contents of malondialdehyde(MDA)and reactive oxygen species(ROS),superoxide dismutase(SOD),and lactate dehydrogenase(LDH)in renal tissues were detected by kits,the expressions of PI3K/Akt/mTOR signaling pathway and apoptotic proteins in renal tissues were detected by Western blot method,the expressions of autophagy-related proteins and ubiquitylated protein mRNA in renal tissues were detected by RT-PCR method.Results The levels of BUN,Cr,IL-1β,IL-6,and IL-18 were significantly lower in the high-dose and low-dose groups of PNS than those in the model group(all P<0.05),and the pathological damage was significantly milder than that in the model group;the contents of MDA,ROS,and LDH,and the relative expressions of p-mTOR,cysteine aspartase-3(Caspase-3),and Bcl-2-associated protein X(Bax)in renal tissues of the Panax notoginseng total saponin low-dose group was significantly lower than those of the model group(all P<0.05),and the contents of SOD and the relative expressions of p-Akt

关 键 词：肾缺血再灌注 三七总皂苷 磷酸肌醇3激酶 蛋白激酶B 哺乳动物雷帕霉素靶蛋白 自噬 泛素蛋白 

分 类 号：R-332[医药卫生]