普伐他汀对子痫前期样小鼠模型中Rheb表达的影响  

Changes of Rheb gene and protein expression in preeclampsia-like mouse model treated with pravastatin

作　　者：淮静 杨孜[1] 易雁鸿[1] 王广娇 Huai Jing;Yang Zi;Yi Yanhong;Wang Guangjiao(Department of Obstetrics and Gynecology,Peking University Third Hospital,Beijing 100191,China)

机构地区：[1]北京大学第三医院妇产科,100191

出　　处：《中华妇产科杂志》2019年第1期38-43,共6页Chinese Journal of Obstetrics and Gynecology

基　　金：国家自然科学基金(81370723).

摘　　要：目的观察普伐他汀(Pra)干预的子痫前期(PE)样小鼠模型中脑Ras同源蛋白(Rheb)的表达变化,探讨Pra对子痫前期样小鼠模型中哺乳动物雷帕霉素靶蛋白(mTOR)信号通路的抑制是否通过Rheb调节。方法将C57BL/6J孕鼠随机分组,于妊娠7~18 d每天注射亚硝基左旋精氨酸甲酯(L-NAME)建立PE样模型,并设立生理盐水对照(Con);于妊娠第8天起再分别每天给予Pra(PE+Pra组、Con+Pra组)或生理盐水(PE+N组、Con+N组),每组8只。于妊娠18 d收集孕鼠的肝脏及胎盘组织;采用蛋白印迹法、实时荧光定量PCR技术和免疫组化法检测组织中Rheb基因和蛋白的表达水平,并比较各组孕鼠肝脏及胎盘组织中Rheb的表达水平变化。结果(1)蛋白印迹法检测结果:PE+N组(肝脏:0.706±0.123;胎盘:0.866±0.128)及Con+N组(肝脏:0.732±0.123;胎盘:0.909±0.097)孕鼠的肝脏和胎盘组织中Rheb蛋白的表达水平,分别比较,差异均无统计学意义(P均>0.05);PE+Pra组(肝脏:0.669±0.134;胎盘:0.940±0.221)与PE+N组孕鼠比较,差异也均无统计学意义(P均>0.05)。(2)实时荧光定量PCR检测结果:PE+N组(肝脏:1.026±0.480;胎盘:1.102±0.361)与Con+N组(肝脏:1.058±0.389;胎盘:1.067±0.400)孕鼠的肝脏和胎盘组织中Rheb mRNA的表达水平,分别比较,差异均无统计学意义(P均>0.05);PE+Pra组(肝脏:0.735±0.356;胎盘:0.822±0.304)与PE+N组比较,差异也均无统计学意义(P均>0.05)。(3)免疫组化结果:Rheb蛋白在各组孕鼠肝脏和胎盘组织的表达部位无明显差异,PE+N组与Con+N组、PE+Pra组与PE+N组的Rheb蛋白表达水平分别比较,差异均无统计学意义(P均>0.05)。结论Pra在PE样小鼠模型中抑制mTOR信号通路的作用,可能与Rheb基因和蛋白的表达水平无关。ObjectiveTo explore whether pravastatin (Pra) inhibits mammalian target of rapamycin (mTOR) signal pathway by regulating Ras homolog enriched in brain (Rheb) protein through the comparison of gene and protein expression changes of Rheb in liver and placenta in preeclampsia (PE)-like mouse model treated with Pra.MethodsC57BL/6J pregnant mice were randomly divided into two groups. The PE group was established by injecting N-nitro-L-arginine methyl ester (L-NAME) daily at gestational 7-18 days, saline was injected as contol group (Con);then giving mice Pra (PE+Pra, Con+Pra group, n=8) or normal saline (PE+N, Con+N group, n=8) every day from the 8th gestational day of pregnancy. The maternal liver and placenta tissues were collected on the 18th day of pregnancy. Western blot, real-time quantitative PCR and immunohistochemistry were used to compare the levels of Rheb protein and mRNA expression in the liver and placenta.Results(1)The results of western blot: there were no significant differences in Rheb protein expression between PE+N group (liver: 0.706±0.123;placenta: 0.866±0.128) and Con+N group (liver: 0.732±0.123;placenta: 0.909±0.097) , and the differences between PE+Pra group (liver: 0.669±0.134;placenta: 0.940±0.221) and PE+N group were not significant either in liver or in placenta (all P>0.05). (2) The results of real-time quantitative PCR: when PE+N group (liver: 1.026±0.480;placenta: 1.102±0.361) compared with Con+N group (liver: 1.058±0.389;placenta: 1.067±0.400) , PE+Pra group (liver: 0.735±0.356;placenta: 0.822±0.304) compared with PE+N group, there were no significant differences either in liver or in placenta (all P>0.05). (3) The results of immunohistochemistry: Rheb protein expression did not change significantly in maternal liver and placenta, there were no significant differences in protein expression levels between PE+N group and Con+N group, and between PE+Pra group and PE+N group (all P>0.05). ConclusionThe inhibition of Pra on mTOR signaling pathway in some PE-like model may be ind

关 键 词：先兆子痫 普伐他丁 富含脑蛋白Ras同源蛋白 疾病模型 动物 

分 类 号：R714.244[医药卫生—妇产科学] R-332[医药卫生—临床医学]