Distribution and expression of α-actin-4 in puromycin aminonueleoside injured mouse podocyte cell line
10.3760/cma.j.issn.2095-428X.2014.17.005
- VernacularTitle:嘌呤霉素损伤小鼠肾小球足细胞系中足细胞α-辅肌动蛋白4的表达与分布
- Author:
Huiyang WANG
;
Li YU
;
Xiaoyan LI
;
Shengyou YU
;
Yao ZHANG
;
Zhihong HAO
- Publication Type:Journal Article
- Keywords:
Podocyte;
α-actin-4;
Puromycin aminonueleoside
- From:
Chinese Journal of Applied Clinical Pediatrics
2014;29(17):1296-1300
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe the expression and distribution of α-actin-4 mRNA through puromycin aminonueleoside(PAN) injury podocyte,and discuss the relation between α-actin-4 and podocyte damage.Methods Podocytes were cultured in vitro,and 2 groups were set up:control group and PAN stimulation group.The control group was cultured with concentration of 100 mL/L FBS RPMI 1640 nutrient solution culture,while the PAN group was cultivated to PAN(50 mg/L) treatment,and cell morphology,extraction of total RNA of α-actin-4 were observed in 8 h,24 h and 48 h.The podocyte morphology was observed and pictures were taken through phase-contrast microscope,then the differences of morphology and areas between the 2 groups were analyzed.The distribution and mRNA expression of α-actin-4 were detected by indirect immunocyto-fluorescence and real-time quantitative PCR,respectively.Results The well-developed podocyte arborization was formed after the in vitro induction,and the PAN treatment led to the podocyte foot process retraction and effacement together with the mouse podocyte cell line shrinkage and the loss of cell contact.The above time point α-actin-4 mRNA expressions between the 2 groups were compared,and there was no significant difference in 8 h (P ＞ 0.05),but significant difference was found in 24 h,48 h,α-actin-4 higher mRNA expression,with statistical significance(all P ＜0.01).α-actin-4 in the control group had thin filaments evenly distributed in the cytoplasm,but a radioactive distribution in foot process.In the experimental group,α-actin-4 pressure silk fiber was shorter,with disordered arrangement,and PAN stimulus after 24 h,α-actin-4 distribution in cytoplasm was decreased significantly,while cytoplasmic distribution was missing after 48 h.Conclusions The abnormal of distribution and mRNA expression of α-actin-4 is time-related to the PAN injury podocyte,and α-actin-4 is an important part of podocyte damage mechanism.
