CAJ | 학술논문

目的：探讨表面活性蛋白-A（SP-A）的表达改变对脂多糖（LPS）诱导的人肾小管近曲上皮细胞（HK-2细胞）白介素-6（IL-6）表达的影响及机制。方法培养HK-2细胞，ELISA方法检测不同浓度的LPS（0、0．1、1、2、5、10 mg／L）作用8 h及5 mg／L的LPS于不同时间（0、2、4、8、16、24 h）作用HK-2细胞后IL-6蛋白表达的变化；应用脂质体转染法将SP-A SiRNA转染入HK-2细胞，筛选稳定转染细胞株，采用5 mg／L的LPS刺激SP-A SiRNA稳定转染的HK-2细胞8 h，ELISA方法检测细胞上清中IL-6的分泌量，Western印迹检测细胞NF-κB p65蛋白的表达。结果应用1、2、5、10 mg／L的LPS刺激HK-2细胞8 h后，IL-6蛋白表达水平较0、0．1 mg／L的LPS刺激后显著升高（P＜0．05）；同时，应用5 mg／L的LPS作用HK-2细胞4、8、16、24 h后，IL-6蛋白的表达水平较5 mg／L的LPS作用0、2 h显著升高（P＜0．05）。SP-A SiRNA转染的HK-2细胞经LPS刺激后，其NF-κBP65和IL-6蛋白表达较未转染经LPS刺激细胞明显升高（P＜0．05）。结论 SP-A可能通过抑制NF-κB p65的表达进而下调LPS诱导的HK-2细胞IL-6的表达，在脓毒症急性肾损伤时发挥保护作用。
목적：탐토표면활성단백-A（SP-A）적표체개변대지다당（LPS）유도적인신소관근곡상피세포（HK-2세포）백개소-6（IL-6）표체적영향급궤제。방법배양HK-2세포，ELISA방법검측불동농도적LPS（0、0．1、1、2、5、10 mg／L）작용8 h급5 mg／L적LPS우불동시간（0、2、4、8、16、24 h）작용HK-2세포후IL-6단백표체적변화；응용지질체전염법장SP-A SiRNA전염입HK-2세포，사선은정전염세포주，채용5 mg／L적LPS자격SP-A SiRNA은정전염적HK-2세포8 h，ELISA방법검측세포상청중IL-6적분비량，Western인적검측세포NF-κB p65단백적표체。결과응용1、2、5、10 mg／L적LPS자격HK-2세포8 h후，IL-6단백표체수평교0、0．1 mg／L적LPS자격후현저승고（P＜0．05）；동시，응용5 mg／L적LPS작용HK-2세포4、8、16、24 h후，IL-6단백적표체수평교5 mg／L적LPS작용0、2 h현저승고（P＜0．05）。SP-A SiRNA전염적HK-2세포경LPS자격후，기NF-κBP65화IL-6단백표체교미전염경LPS자격세포명현승고（P＜0．05）。결론 SP-A가능통과억제NF-κB p65적표체진이하조LPS유도적HK-2세포IL-6적표체，재농독증급성신손상시발휘보호작용。
Objective To evaluate the effects and mechanisms of surfactant protein A(SPA)onlipopolysaccharide(LPS)induced interleukin6(IL6)expression in human renal tubular epithelial(HK2)cells.Methods Cultured HK2 cells were treated with various concentrations of LPS(0,0.1,1,2,5and 10 mg/L)for 8 h,and 5 mg/L of LPS at different time point(0,2,4,8,16 and 24 h),and the changeof IL6 expression was tested by the ELISA method.Then HK2 cells were transfected with SPA siRNA.Stable transfected HK2 cells were screened and stimulated with LPS(5 mg/L)for 8 h and NFkB P65 expression was evaluated by Western blot.Results The levels of IL6 expression in HK2 cells treated with1,2,5 and 10 mg/L of LPS were higher than these cells treated with 0 and 0.1 mg/L of LPS.After the 5mg/L of LPS treatment for 4,8,16 and 24 h,their IL6 levels IN HK2 cells were higher than these cellstreated for 0 and 2 h.Compared with those nontransfected LPStreated HK2 cells,the expressions of NFkB P65 and IL6 in the transfected cells were elevated significantly(P <0.05).Conclusion By inhibiting the expression of NFkB P6,SPA may downregulate LPSinduced IL6 expression in HK2 cells,which plays a protective role in sepsis and acute kidney injuries.