胆红素和内毒素联合作用对NRK52E细胞缝隙连接功能的影响

Function of gap junction in NRK52E cells exposed to bilirubin combined with lipopolysaccharide

目的:观察胆红素(BR)和内毒素(LPS)联合作用对肾小管上皮细胞(NRK52E)生长及细胞间缝隙连接(GJ)的影响。方法:体外培养NRK52E细胞,不同浓度的BR和LPS联合干预,用MTT测量细胞生长;观察它们对生长融合细胞(有GJ形成)和生长未融合细胞(无GJ形成)集落形成的影响;采用细胞荧光免疫示踪法分析细胞间GJ的功能。结果:BR从17.1μmol/L增加至513μmol/L,可浓度依赖性地增加细胞生长;当BR浓度继续增加时,细胞生长逐渐降低。LPS(10-1 000μg/L)能浓度依赖性地降低NRK52E细胞生长。BR和LPS联合作用下,513μmol/L BR增加100μg/L LPS作用下的细胞生长(P<0.05),而684μmol/L BR降低100μg/L LPS的细胞生长(P<0.05);513μmol/L BR能增加100μg/L LPS作用下GJ传递数目(P<0.05),684μmol/L BR降低100μg/L LPS作用下的GJ传递数目。结论:BR和LPS联合作用时,513μmol/L BR降低LPS的细胞毒性,684μmol/LBR增加LPS的细胞毒性,其改变可能是通过细胞间缝隙连接发挥作用的。

AIM ： To observe the effect of bilirubin （BR） combined with lipopolysaccharide （LPS） on the growth and gap junction of NRK52E cells. METHODS ： NRK52E cells were cultured and treated with different concentrations of bilirubin combined with lipopolysaccharide. The growth of NRK52E cells was measured by MTT method. The tunction of gap junction in NRK52E cells was determined by the method of fluoroimmunoassay. RESULTS ： BR promoted the growth of NRK52E cells in a dose - dependent manner from 17.1 μmol/L to 513 μLmol/L but inhibited it at more than 513 μLmol/L. LPS inhibited the growth of NRK52E cells in a dose - dependent manner from 10 - 1 000 μg/L. Under the condition of BR combined with LPS at 100 μg/L, BR at 513 μmol/L increased the growth of NRK52E cells while BR at 684 μmol/L decreased the growth of the cells （P 〈0. 05）. Combined with LPS at 100 μg/L, BR at 513 μmol/L increased the permeability of the gap junction （P 〈 0. 05 ）, while BR at 684 μmol/L decreased the permeability of the gap junction. CONCLUSION： BR at a low concentration （513 μmol/L） decreases, and BR at a high concentration （684μmol/L） increases the toxicity of LPS on NRK52E cells by affecting the function of gap junctions in the cells.