CAJ | 학술논문

目的：研究热休克蛋白（HSP）60在纳洛酮神经保护中的作用及机制。方法培养小胶质细胞株 BV2，实验分为对照组、阳性对照组（细菌脂多糖，LPS）、实验组（ LPS＋纳洛酮）。利用Western印迹和免疫荧光技术检测不同分组中 HSP60的表达，ELISA技术检测 HSP60的胞外释放。利用人重组HSP60（rhHSP60）作用于BV2细胞，然后用 Western 印迹技术检测不同分组中 Toll 样受体（TLR）-4的表达；收集细胞培养液，用ELISA检测肿瘤坏死因子（ TNF）-α和γ-干扰素（ IFN-γ）的水平。结果与对照组相比，阳性对照组中的 HSP60和 TLR-4表达及TNF-α和 IFN-γ的释放都显著增加；而加入纳洛酮后，实验组中这些因子的水平都明显降低（ P＜0.05）。结论纳洛酮可能通过 HSP60-TLR-4途径阻止小胶质细胞的活化而发挥神经保护作用。
목적：연구열휴극단백（HSP）60재납락동신경보호중적작용급궤제。방법배양소효질세포주 BV2，실험분위대조조、양성대조조（세균지다당，LPS）、실험조（ LPS＋납락동）。이용Western인적화면역형광기술검측불동분조중 HSP60적표체，ELISA기술검측 HSP60적포외석방。이용인중조HSP60（rhHSP60）작용우BV2세포，연후용 Western 인적기술검측불동분조중 Toll 양수체（TLR）-4적표체；수집세포배양액，용ELISA검측종류배사인자（ TNF）-α화γ-간우소（ IFN-γ）적수평。결과여대조조상비，양성대조조중적 HSP60화 TLR-4표체급TNF-α화 IFN-γ적석방도현저증가；이가입납락동후，실험조중저사인자적수평도명현강저（ P＜0.05）。결론납락동가능통과 HSP60-TLR-4도경조지소효질세포적활화이발휘신경보호작용。
Objective To investigate the function and possible mechanism of HSP60 in the neuroprotective effects of naloxone. Methods Culturing BV2 cell line were cultured and devided into control,positive control and experimental groups.The expression of HSP60 in 3 groups was measured by Western blot and immunofluorescence technique.The extracellular release of HSP60 was detected by ELISA. The expression of TLR-4 was measured by Western blot after rhHSP60 in BV2 cell,and the releases of TNF-αand IFN-γwere observed by ELISA.Results Compared with those of control group,the expressions of HSP60,TLR-4,TNF-αand IFN-γin positive control group were significantly increased,and their expressions were decreased in experimental group ( P<0.05).Conclusions Naloxone might through HSP60-TLR-4 signal path to inhibit the activation of microglia and play neuroprotective effects.