CAJ | 학술논문

观察旋毛虫重组热休克蛋白HSP70（rTs-HSP70）对体外培养小鼠巨噬细胞Toll样受体TLR2/4表达的影响。体外常规培养小鼠巨噬细胞系RAW264.7，分别加入不同浓度rTs-HSP70刺激培养24 h，半定量PCR方法检测培养细胞TLR2/4 mRNA表达，流式细胞仪检测表面标志分子CD80和CD86表达。平行设立不加刺激物阴性对照组，细菌脂多糖LPS阳性对照组；同时试验比较5μg·mL-1浓度的HSP70组和HSP70+LPS组（HSP70预刺激后再加入LPS刺激培养12 h）RAW264.7细胞TLR2/4 mRNA表达差异。结果表明，低浓度rTs-HSP70与巨噬细胞活化呈正相关，浓度为5μg·mL-1时，TLR2/4表达水平达到峰值（P<0.05），并可刺激巨噬细胞共刺激分子CD80和CD86高表达，随着HSP70浓度升高（>5μg·mL-1）则对巨噬细胞RAW264.7表现为抑制作用；当对RAW264.7进行旋毛虫HSP70预刺激，可引起免疫抑制效应，抑制LPS对巨噬细胞TLR2/4的活化反应。试验可为旋毛虫相关分子免疫调节机制研究及旋毛虫病防治提供参考。
관찰선모충중조열휴극단백HSP70（rTs-HSP70）대체외배양소서거서세포Toll양수체TLR2/4표체적영향。체외상규배양소서거서세포계RAW264.7，분별가입불동농도rTs-HSP70자격배양24 h，반정량PCR방법검측배양세포TLR2/4 mRNA표체，류식세포의검측표면표지분자CD80화CD86표체。평행설립불가자격물음성대조조，세균지다당LPS양성대조조；동시시험비교5μg·mL-1농도적HSP70조화HSP70+LPS조（HSP70예자격후재가입LPS자격배양12 h）RAW264.7세포TLR2/4 mRNA표체차이。결과표명，저농도rTs-HSP70여거서세포활화정정상관，농도위5μg·mL-1시，TLR2/4표체수평체도봉치（P<0.05），병가자격거서세포공자격분자CD80화CD86고표체，수착HSP70농도승고（>5μg·mL-1）칙대거서세포RAW264.7표현위억제작용；당대RAW264.7진행선모충HSP70예자격，가인기면역억제효응，억제LPS대거서세포TLR2/4적활화반응。시험가위선모충상관분자면역조절궤제연구급선모충병방치제공삼고。
To observe the effect of 70 kDa recombinant heat shock protein from Trichinella spiralis (rTs-HSP70) on expression of TLR2/4 (toll-like receptors) in murine macrophage in vitro. Murine macrophage RAW264.7 cell lines were incubated in twelve-well plates and different concentrations of rTs-HSP70 were added, co-culturing for 24 hours. Then the cells’TLR2/4 expression were tested by semi-quantitative PCR, those of surface markers CD80 and CD86 by flow cytometry. Cells receiving no stimulation were taken as negative control, bacterial lipopolysaccharide (LPS) as positive control. At the same time, comparative studies on incubation with rTs-HSP70 alone, rTs-HSP70 in combination with LPS, were conducted at concentration 5 μg·mL-1 of HSP70. The results indicated that lower concentra-tions of rTs-HSP70 had a positive correlation with macrophages activation, TLR2/4 expression peaked at concentration of 5 μg·mL-1 (P<0.05), and up-regulation of co-stimulatory molecules. when rTs-HSP70 concentration rose above a certain threshold (>5 μg·mL-1), a inhibitory effect on macrophage could be observed. Pre-stimulation of the macrophage with rTs-HSP70 could result in immunosuppression, generating a suppresive effect on LPS in TLR2/4 activation. The study lay the foundation for the mechanisms of helminth molecules immunomodulation and the therapeutic treatment of different trichinellosis.