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肾脏病与透析肾移植杂志

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外泌体miR155调控p38丝裂原活化蛋白激酶信号通路影响心肾综合征大鼠的心肾细胞凋亡

  

  • 出版日期:2019-12-28 发布日期:2020-01-19

Exosome miR155 regulates p38MAPK signaling pathway and affects cardiomyocytnephrocyte apoptosis in rats with cardiorenal syndrome

  • Online:2019-12-28 Published:2020-01-19

摘要: 目的:基于外泌体miR155调控p38丝裂原活化蛋白激酶(p38MAPK)信号通路影响心肾综合征(CRS)大鼠的心肾细胞凋亡探讨CRS的发病机制。
方法:SD大鼠分为空白组、造模3周组、造模4周组、造模5周组、造模6周组,每组10只,以生理盐水或阿霉素腹腔注射造模。心脏超声检测心功能,ELISA 法检测血清氨基末端脑钠肽前体(NTproBNP)、肾损伤分子1,全自动生化仪检测血清肌酐、尿素氮,EXOQuick试剂盒提取血浆外泌体,用ELISA试剂盒对血浆外泌体定量检测,RTPCR 法检测心肾组织p38MAPK mRNA、心肾组织miR155、外泌体miR155的表达,WesternBlot法检测心肾组织p38MAPK及磷酸化p38MAPK(pp38MAPK)的表达,HE染色观察组织形态学改变,TUNEL法检测心肾细胞凋亡率。
结果:造模后各组大鼠心肾功能降低;与空白组比较,造模4周组、5周组心脏pp38MAPK表达增高(P<005),6周组pp38MAPK表达增高(P<001); 造模4周组、5周组、6周组肾脏pp38MAPK表达增高(P<001);造模3周组、4周组、5周组、6周组心脏、肾脏细胞凋亡率增高(P<001);造模 4周组、5周组、6周组血浆外泌体含量降低(P<001);造模3周组、4周组、5周组、6周组外泌体miR155表达降低,心肾组织miR155表达降低(P<001)。
结论:pp38 MAPK在CRS大鼠心肾细胞凋亡中起着关键作用,且外泌体miR155可能对其有着调控作用。

关键词: 心肾综合征, 外泌体, p38丝裂原活化蛋白激酶, 细胞凋亡

Abstract: Objective:To explore the pathogenesis of cardiorenal syndrome(CRS) based on the effect of exosome miR155 on apoptosis of cardiorenal cells in rats with CRS by regulating p38MAPK signaling pathway.
Methodology:SpragueDawley rats were randomly divided into control group, model group for 3 weeks, model group for 4 weeks, model group for 5 weeks, and model group for 6 weeks.Each group received 10 rats with intraperitoneal injection of normal saline or doxorubicin.Histomorphological changes were observed by HE staining.Cardiac function was detected by echocardiography.Serum NTpro BNP and KIM1 were detected by ELISA.Urea nitrogen and serum creatinine were detected by automatic biochemical analyzer.EXO Quick kit was used to extract plasma exosomes and detect them quantitatively.RTPCR, Westernblot  were used to detect expressions of p38MAPK mRNA, miR155, p38MAPK and pp38MAPK in heart and kidney tissue.The apoptosis of heart and kidney was detected by TUNEL.
Results:Cardiac and renal function of rats in each group decreased after model establishment.Compared with the blank group,PP38 in the heart increased in the 4week group and the 5week group (P<005),6week group (P<001).Compared with the blank group,the kidney PP38 increased in the 4week group,the 5week group and the 6week group (P<001).Compared with the blank group,the apoptotic rate of heart and kidney cells increased in the 3week group,the 4week group,the 5week group and the 6week group (P<001); the plasma exosome content decreased in the 4week group,the 5week group and the 6week group (P<001); the exosome miR155 decreased in the 3week group,the 4week group,the 5week group and the 6week group (P<001),and the miR155 decreased in the heart and kidney tissue (P<001).
Conclusion:
The phosphorylation of p38 MAPK plays a key role in the apoptosis of cardiac and renal cells in CRS rats,and the exosome miR155 may play a regulatory role.