摘要
目的:探讨C3a受体(C3a receptor,C3aR)在人壁层肾祖细胞(CD24^(+)CD133^(+)PEC)向足细胞分化中的作用及其机制。方法:分离和培养原代人CD24^(+)CD133^(+)PEC,诱导分化为足细胞并检测C3aR在分化过程中的表达。构建C3a分泌性表达慢病毒,感染CD24^(+)CD133^(+)PEC,获得C3a过表达细胞株(C3a过表达组)。检测C3a过表达组、空载体组和未感染组细胞的活力、分化情况及E-cadherin、p-ERK1/2的表达。结果:成功分离出CD24^(+)CD133^(+)PEC并诱导其分化为表达Wilms肿瘤蛋白1(Wilms tumor protein 1,WT1)的足细胞,C3aR在分化过程中表达显著降低。C3a过表达组的细胞活力较空载体组和未感染组显著增强(P<0.01);诱导分化第2天,C3a过表达组细胞WT1的表达较空载体组和未感染组显著降低(P<0.01),E-cadherin及p-ERK1/2的表达较空载体组和未感染组显著升高(P<0.01)。结论:C3aR参与了人CD24^(+)CD133^(+)PEC向足细胞分化的过程,C3a过表达促进CD24^(+)CD133^(+)PEC的增殖并抑制其向足细胞分化,可能是通过ERK1/2信号通路发挥作用的。
AIM:To investigate the role of C3a receptor(C3aR)in the differentiation of parietal renal progenitor(CD24^(+)CD133^(+)parietal epithelial cell,CD24^(+)CD133^(+)PEC)into podocyte.METHODS:The primary CD24^(+)CD133^(+)PECs were isolated,cultured and differentiated into podocytes,and the expression of C3aR was detected during the differentiation process.C3a secretory expression lentiviruses were constructed and infected to CD24^(+)CD133^(+)PECs,and C3a over-expression cell line was obtained(C3a over-expression group).The viability and differentiation capacities and the protein levels of E-cadherin and p-ERK1/2 of cells in C3a over-expression group,empty vector group and uninfected group were determined.RESULTS:CD24^(+)CD133^(+)PECs were successfully isolated and differentiated into podocytes expressing Wilms tumor protein 1(WT1).The expression of C3aR significantly reduced during the differentiation process.The cell viability in C3a over-expression group was significantly higher than that in uninfected group and empty vector group(P<0.01).On the second day of differentiation,WT1 protein expression in C3a over-expression group was significantly lower than that in uninfected group and empty vector group(P<0.01),E-cadherin and p-ERK1/2 protein levels were significantly higher than that in uninfected group and empty vector group(P<0.01).CONCLUSION:C3aR is involved in the differentiation of CD24^(+)CD133^(+)PEC into podocyte and the over-expression of C3a promotes its proliferation and inhibits its differentiation into podocyte,which might work through the ERK1/2 signaling pathway.
作者
杨静
叶璐霞
王莎莎
YANG Jing;YE Lu-xia;WANG Sha-sha(Department of Nephrology,The First Affiliated Hospital of Soochow University,Soochow 215000,China;Taizhou Hospi-tal of Zhejiang Province Affiliated to Wenzhou Medical University,Wenzhou 317000,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2021年第12期2228-2236,共9页
Chinese Journal of Pathophysiology
基金
台州市科技计划项目(No.1801ky02)。
