摘要
目的探讨微RNA(miRNA)-181靶向磷酸酶张力蛋白基因(PTEN)调控磷脂酰肌醇3-激酶/蛋白质丝氨酸苏氨酸激酶(PI3K/Akt)信号通路在高尿酸血症大鼠肾损伤中的作用。方法选择雄性Wistar大鼠40只,随机分为对照组10只、模型组10只、阴性对照组10只和miRNA-181抑制组10只,检测大鼠血清尿酸、肌酐、尿素氮;采用苏木精-伊红(HE)染色,光镜下观察各组大鼠肾脏组织病理形态学改变;实时荧光定量(qRT)-PCR检测各组大鼠肾组织miRNA-181和PTEN、PI3K、Akt mRNA表达水平;蛋白质印迹法检测各组大鼠肾组织PTEN、PI3K、Akt、磷酸化(p)-Akt蛋白表达水平;双荧光素酶报告基因实验验证miRNA-181与PTEN的靶向关系。多组间比较采用单因素方差分析,方差齐,进一步组间两两比较采用LSD-t检验;若方差不齐,进一步组间两两比较采用Tamhane′s T2检验;2组间比较采用独立样本t检验。结果与对照组尿酸(135±21)mmol/L、肌酐(27.8±2.1)μmol/L、尿素氮(6.8±0.5)μmol/L相比,模型组和阴性对照组大鼠尿酸[(213±28)mmol/L,(214±23)mmol/L;肌酐(49.2±2.3)μmol/L,(48.6±2.2)μmol/L;尿素氮(11.5±2.7)μmol/L,(11.7±2.5)μmol/L]含量显著升高(F尿酸=26.739,F肌酐=259.055,F尿素氮=12.921,均P<0.05);与阴性对照组相比,miRNA-181抑制组大鼠尿酸(169±21)mmol/L、肌酐(33.7±1.8)μmol/L、尿素氮(9.1±1.7)μmol/L含量降低(LSD-t尿酸=4.356,LSD-t肌酐=15.773,LSD-t尿素氮=2.858,均P<0.05)。模型组和阴性对照组大鼠肾组织miRNA-181表达水平(1.88±0.16、1.84±0.18)显著高于对照组(0.53±0.08)(F=193.554,P<0.05),而PTEN蛋白(0.18±0.02、0.16±0.02)和mRNA表达水平(0.48±0.08、0.44±0.07)均低于对照组(1.27±0.06、1.27±0.16)(F蛋白表达水平=515.116,FmRNA表达水平=141.470,均P<0.05);抑制miRNA-181后,大鼠肾组织肾组织miRNA-181表达水平(1.35±0.58)显著降低(LSD-t=10.341,P<0.05),PTEN蛋白(0.84±0.05)和mRNA表达水平(0.90±0.08)均升高(LSD-t蛋白表达水平=20.471,Tamhan
Objective To investigate the role of miRNA-181 targeting phosphatase and tensin homologue deleted on chromosome ten(PTEN)in regulating phosphatidylinositol-3-kinase/Akt signaling pathway(PI3K/Akt)signaling pathway in renal injury of hyperuricemia rats.Methods Forty male Wistar rats were randomly divided into control group,model group,negative control group and miRNA-181 inhibition group.Their serum uric acid,creatinine and urea nitrogen were tested.HE staining was used to observe the renal histopathological changes in each group.The expression of miRNA-181,PTEN,PI3K and Akt mRNA in renal tissue of rats in each group was detected by quantitative real time-polymerase chain reaction(qRT-PCR).Western blotting analysis of PTEN,PI3K,Akt and p-Akt protein expression in renal tissue of rats in each group.The targeting relationship between miRNA-181 and PTEN was confirmed by double luciferase reporter gene experiment.One-way analysis of variance(ANOVA)was used for the comparison between multiple groups,with the same variance.LSD-t test was used for further comparison between the two groups.If the variance was not the same,Tamhane's T2 test was used for further comparison between the two groups.Independent sample t-test was used to compare between the two groups.Results Compared with the control group(135±21)mmol/L;(27.8±2.1)μmol/L;(6.8±0.5)μmol/L,the contents of uric acid[(213±28)mmol/L,(214±23)mmol/L,creatinine(49.2±2.3)μmol/L,(48.6±2.2)μmol/L and urea nitrogen(11.5±2.7)μmol/L;(11.7±2.5)μmol/L]in the model group and the negative control group were significantly increased(Furic acid=26.739,Fcreatinine=259.055,Furea nitrogen=12.921,all P<0.05);compared with the nega-tive control group,the contents of uric acid(169±21)mmol/L,creatinine(33.7±1.8)μmol/L and urea nitrogen(9.1±1.7)μmol/L in the miRNA-181 inhibition group were decreased(LSD-turic acid=4.356,LSD-tcreatinine=15.773,LSD-turea nitrogen=2.858,all P<0.05).The expression level of miRNA-181 in renal tissue of the model group and the negative control
作者
杜彭
陈明
兰颖
杨云华
邓长财
Du Peng;Chen Ming;Lan Ying;Yang Yunhua;Deng Changcai(Department of Renal Rheumatology,Tianjin 4th Central Hospital,Tianjin 300140,China;Department of Rheumatology Immunology,General Hospital of Tianjin Medical University,Tianjin 300052,China)
出处
《中华风湿病学杂志》
CAS
CSCD
北大核心
2020年第8期530-535,I0002,共7页
Chinese Journal of Rheumatology
