全氟辛酸对HK-2细胞的毒性及对α-酮戊二酸盐受体1表达的影响

Toxicity of perfluorooctanoic acid to HK-2 cells and its effect on the expression of oxogluatarate receptor 1

目的:研究全氟辛酸(perfluorooctanoic acid,PFOA)对HK-2细胞的毒性,及其对α-酮戊二酸盐受体1(oxoglutarate receptor 1,OXGR1)表达的影响。方法:采用MTT法检测不同浓度(0、0.1、1、10、100、1 000μmol/L)PFOA对HK-2细胞增殖的影响。RT-PCR法检测0、100、300μmol/L PFOA对HK-2细胞中OXGR1、琥珀酸盐受体1(succinate receptor 1,SUCNR1)、缺氧诱导因子1α(hypoxia-inducible factor 1α,HIF-1α)mRNA表达的影响。当PFOA为300或0μmol/L时,采用MTT法检测不同浓度(0、1、10、100、1 000、10 000μmol/L)OXGR1配基α-酮戊二酸钠(α-ketoglutarate sodium,AKG)对HK-2细胞增殖的影响。流式细胞术检测300μmol/L PFOA和100μmol/L AKG共同作用HK-2细胞48 h后细胞的凋亡情况。结果:在培养24 h条件下,PFOA达1 000μmol/L时可表现出毒性作用(P<0.01)。在培养48或72 h条件下,PFOA≥100μmol/L时表现出毒性作用(P<0.01)。RT-PCR结果显示,PFOA可上调HK-2细胞中OXGR1 mRNA表达(P<0.01),但对SUCNR1及HIF-1αmRNA表达无明显影响(P>0.05)。MTT及流式细胞术检测结果显示,AKG与PFOA可协同作用抑制HK-2细胞增殖(P<0.01)并诱导细胞凋亡(P<0.05)。结论:PFOA对HK-2细胞具有增殖抑制作用,并可上调细胞OXGR1 mRNA表达;PFOA可诱发HK-2细胞凋亡;AKG可与PFOA产生协同作用,该协同作用可能与OXGR1有关。

OBJECTIVE：To explore the toxicity of perfluorooctanoic acid （PFOA） to HK-2 cells,and its effect on the expression of oxoglutarate receptor 1 （OXGR1） in HK-2 cells. METHODS：The effect of different concentrations of PFOA on HK-2 cells proliferation was examined by MTT assay. The effect of PFOA （0,100,300μmol/L） on mRNA expression of OXGR1,succinate receptor 1 （SUCNR1） and hypoxia-inducible factor 1α（HIF-1α）in HK-2 cells were evaluated by RT-PCR. The effect of different concentrations （0,1, 10,100,1 000,10 000μmol/L） ofα-ketoglutarate sodium （AKG） on HK-2 cell proliferation with or without PFOA （300μmol/L） was assayed by MTT method. Apoptoses of HK-2 cells after 48 h of culture in medium with PFOA （300 μmol/L） and AKG （100μmol/L） were examined by flow cytometry. RESULTS：1 000μmol/L of PFOA exhibited toxicity after 24 h of culture （P〈0.01）. PFOA（≥100 μmol/L） exhibited toxicity after 48 h or 72 h of culture （P〈0.01）. RT-PCR showed that PFOA upregulated OXGR1 mRNA expression in HK-2 cells,but it had no effects on the expressions of SUCNR1 and HIF-1αmRNA. Interaction of AKG and PFOA inhibited proliferation of HK-2 cells （P〈0.001） and induced apoptosis （P〈0.05）, which were measured by MTT and flow cytometry. CONCLUSION： PFOA could inhibit the proliferation of HK-2 cells,and upregulate mRNA expression of OXGR1. PFOA could induce apoptosis of HK-2 cells;interaction of AKG and PFOA could also induce apoptosis,and whether this interaction is OXGR1 dependant remains to be further verified.