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目的 :探讨Oatp3在PFOS诱导的支持细胞毒性中的作用。方法 :体外分离纯化原代大鼠支持细胞,通过细胞毒性实验,观察PFOS对原代支持细胞生长的影响;利用Oatp3 si RNA构建Oatp3基因敲减模型,观察Oatp3在PFOS诱导的支持细胞毒性中的作用,并采用Western blot法检测PFOS对Oatp3蛋白表达的影响。结果:0~30μmol/L剂量的PFOS对大鼠支持细胞生长无明显影响(P>0.05);当PFOS剂量增至40μmol/L时,对支持细胞毒性作用明显(P<0.01),PFOS毒作用的IC50值约为45.6μmol/L。40μmol/L的PFOS可明显诱导Oatp3蛋白的表达(P<0.01),联合Oatp3 si RNA处理后,其诱导作用明显抑制(P<0.01)。与转染试剂对照组(Mock)相比,加入40μmol/L的PFOS后,支持细胞存活率明显降低(P<0.01),联合Oatp3 si RNA处理后,PFOS诱导的支持细胞毒性明显被抑制(P<0.05或P<0.01)。结论:Oatp3很可能参与PFOS诱导支持细胞损伤。
Objective: To investigate the role of Oatp3 in PFOS-induced cytotoxicity. Methods: Primary cultured rat spinal cord cells were isolated and purified in vitro. The effects of PFOS on the growth of primary cells were observed by cytotoxicity assay. Oatp3 gene knockdown model was constructed by Oatp3 si RNA. Oatp3 expression was observed in PFOS-induced cytotoxicity The effect of PFOS on the expression of Oatp3 protein was detected by Western blot. Results: PFOS at 0 ~ 30μmol / L had no significant effect on the growth of rat supportive cells (P> 0.05). When the dose of PFOS was increased to 40μmol / L, the cytotoxicity to supportive cells was significant (P <0.01) The IC50 value is about 45.6 μmol / L. Oatp3 protein expression was significantly induced by 40μmol / L PFOS (P <0.01). Oatp3 si RNA treatment inhibited the expression of Oatp3 protein significantly (P <0.01). Survival rate of support cells was significantly decreased (P <0.01) when treated with 40μmol / L PFOS compared with transfection reagent control group (Mock). After treated with Oatp3siRNA, the cytotoxicity of PFOS was significantly inhibited (P <0.05 or P <0.01). Conclusion: Oatp3 is likely involved in PFOS-induced cell injury.