摘要
目的探讨亚低温对肾脏缺血-再灌注损伤(IRI)的作用,以及RNA结合基序蛋白3(RBM3)及其下游效应分子在这一过程中的表达情况。方法健康SD雄性大鼠18只,随机分为正常对照(NC)组、IRI组和亚低温预处理(MHP)组,每组6只。通过检测血清肌酐水平评价肾功能,苏木素-伊红(HE)染色检测肾脏组织损伤情况,蛋白质印迹法检测肾脏组织的RBM3、Yes相关蛋白1(YAP1)、核因子E2相关因子2(Nrf2)、B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)蛋白相对表达量,免疫组织化学染色进一步检测RBM3及YAP1蛋白的表达情况,采用脱氧核糖核酸末端转移酶介导的dUTP缺口末端标记(TUNEL)法检测肾脏组织细胞凋亡情况,通过测定丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性检测肾脏组织氧化应激水平。结果与NC组相比较,IRI组和MHP组血清肌酐水平、肾脏组织病理损伤评分均升高,RBM3、YAP1和Nrf2蛋白表达水平均升高,Bcl-2/Bax比值均降低,细胞凋亡率均升高,MDA含量均升高,SOD活性均下降,差异均有统计学意义(均为P<0.05);与IRI组相比较,MHP组血清肌酐水平、肾脏组织病理损伤评分均降低,RBM3、YAP1和Nrf2蛋白表达水平均升高,Bcl-2/Bax比值升高,细胞凋亡率降低,MDA含量降低,SOD 活性升高,差异均有统计学意义(均为P<0.05)。结论 亚低温对肾脏IRI 具有保护作用,可减轻IRI 所致细胞凋亡和氧化应激损伤,其机制可能为亚低温上调RBM3 及其下游效应分子YAP1 和Nrf2 的表达水平。
Objective To evaluate the effect of mild hypothermia on the renal ischemia-reperfusion injury(IRI),and the expression profile of RNA-binding motif protein 3(RBM3)and its downstream effector molecules during this process.Methods Eighteen healthy SD male rats were randomly divided into the normal control(NC)group,IRI group and mild hypothermia pretreat(MHP)group,with 6 rats in each group.Serum creatinine level was measured to evaluate the renal function.Hematoxylin-eosin(HE)staining was performed to assess the renal tissue injury.Western blot was used to determine the relative expression levels of RBM3,Yes-associated protein 1(YAP1),nuclear factor E2-related factor 2(Nrf2),B cell-lymphoma-2(Bcl-2)and Bcl-2-associated X protein(Bax)in the kidney tissues.Immunohistochemical staining was employed to further detect the expression levels of RBM3 and YAP1 proteins.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)assay was adopted to detect the cell apoptosis of kidney tissues.Malondialdehyde(MDA)content and superoxide dismutase(SOD)activity were evaluated to determine the oxidative stress level of kidney tissues.Results Compared with the NC group,the serum creatinine level,the pathological injury score of kidney tissues and the expression levels of RBM3,YAP1 and Nrf2 proteins were significantly up-regulated,the Bcl-2/Bax ratio was considerably lower,the apoptosis rate was remarkably elevated,the MDA content was significantly increased and the SOD activity was dramatically reduced in the IRI and MHP groups(all P<0.05).Compared with the IRI group,the serum creatinine level and the pathological injury score of kidney tissues were significantly decreased,the expression levels of RBM3,YAP1 and Nrf2 proteins were significantly up-regulated,the Bcl-2/Bax ratio was considerably higher,the apoptosis rate was significantly decreased,the MDA content was significantly decreased and the SOD activity was considerably elevated in the MHP group(all P<0.05).Conclusions Mild hypothermia may exert protective effect upon renal IRI and it could alleviate cell apoptosis and oxidative stress injury induced by IRI,probably by up-regulating the expression level of RBM3 and its downstream effector molecules of YAP1 and Nrf2.
作者
宋克芹
肖琦
肖建生
罗凯锋
Song Keqin;Xiao Qi;Xiao Jiansheng;Luo Kaifeng(Department of Organ Transplantation,the First Affiliated Hospital of Nanchang University,Nanchang 330006,China)
出处
《器官移植》
CAS
CSCD
北大核心
2021年第5期571-578,共8页
Organ Transplantation
基金
江西省青年科学基金资助项目(20192BAB215012)
江西省卫生健康委科技计划(20203206)。