熱休克蛋白60在納洛酮神經(jīng)保護(hù)中的作用
發(fā)布時(shí)間：2018-08-05 15:33
【摘要】：目的研究熱休克蛋白(HSP)60在納洛酮神經(jīng)保護(hù)中的作用及機(jī)制。方法培養(yǎng)小膠質(zhì)細(xì)胞株BV2,實(shí)驗(yàn)分為對(duì)照組、陽(yáng)性對(duì)照組(細(xì)菌脂多糖,LPS)、實(shí)驗(yàn)組(LPS+納洛酮)。利用Western印跡和免疫熒光技術(shù)檢測(cè)不同分組中HSP60的表達(dá),ELISA技術(shù)檢測(cè)HSP60的胞外釋放。利用人重組HSP60(rh HSP60)作用于BV2細(xì)胞,然后用Western印跡技術(shù)檢測(cè)不同分組中Toll樣受體(TLR)-4的表達(dá);收集細(xì)胞培養(yǎng)液,用ELISA檢測(cè)腫瘤壞死因子(TNF)-α和γ-干擾素(IFN-γ)的水平。結(jié)果與對(duì)照組相比,陽(yáng)性對(duì)照組中的HSP60和TLR-4表達(dá)及TNF-α和IFN-γ的釋放都顯著增加;而加入納洛酮后,實(shí)驗(yàn)組中這些因子的水平都明顯降低(P0.05)。結(jié)論納洛酮可能通過(guò)HSP60-TLR-4途徑阻止小膠質(zhì)細(xì)胞的活化而發(fā)揮神經(jīng)保護(hù)作用。
[Abstract]:Objective to study the role and mechanism of heat shock protein (HSP) 60 (HSP60) in the neuroprotection of naloxone. Methods microglia cell line BV2 was cultured and divided into control group, positive control group (lipopolysaccharide) and LPS naloxone group. The expression of HSP60 in different groups was detected by Western blot and immunofluorescence. The extracellular release of HSP60 was detected by Elisa. Human recombinant HSP60 (rh HSP60) was used to treat BV2 cells, then the expression of Toll like receptor (TLR) 4 was detected by Western blotting, and the levels of (TNF) 偽 and IFN- 緯 were detected by ELISA. Results compared with the control group, the expression of HSP60 and TLR-4 and the release of TNF- 偽 and IFN- 緯 in the positive control group were significantly increased, but the levels of these factors in the experimental group were significantly decreased after the addition of naloxone (P0.05). Conclusion Naloxone may play a neuroprotective role by blocking the activation of microglia through HSP60-TLR-4 pathway.
【作者單位】： 寧夏醫(yī)科大學(xué)寧夏顱腦疾病重點(diǎn)實(shí)驗(yàn)室;銀川市第一人民醫(yī)院高壓氧科;
【基金】：國(guó)家自然科學(xué)基金(No.31260243)
【分類號(hào)】：R96
本文編號(hào)：2166224
[Abstract]:Objective to study the role and mechanism of heat shock protein (HSP) 60 (HSP60) in the neuroprotection of naloxone. Methods microglia cell line BV2 was cultured and divided into control group, positive control group (lipopolysaccharide) and LPS naloxone group. The expression of HSP60 in different groups was detected by Western blot and immunofluorescence. The extracellular release of HSP60 was detected by Elisa. Human recombinant HSP60 (rh HSP60) was used to treat BV2 cells, then the expression of Toll like receptor (TLR) 4 was detected by Western blotting, and the levels of (TNF) 偽 and IFN- 緯 were detected by ELISA. Results compared with the control group, the expression of HSP60 and TLR-4 and the release of TNF- 偽 and IFN- 緯 in the positive control group were significantly increased, but the levels of these factors in the experimental group were significantly decreased after the addition of naloxone (P0.05). Conclusion Naloxone may play a neuroprotective role by blocking the activation of microglia through HSP60-TLR-4 pathway.
【作者單位】： 寧夏醫(yī)科大學(xué)寧夏顱腦疾病重點(diǎn)實(shí)驗(yàn)室;銀川市第一人民醫(yī)院高壓氧科;
【基金】：國(guó)家自然科學(xué)基金(No.31260243)
【分類號(hào)】：R96
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