CAJ | 학술논문

目的本实验对不同毒力结核杆菌的衍生蛋白(PPD)对人巨噬细胞(THP-1)的影响及其与TNF-αt、IL-1 β及IL-10的差异性进行研究.方法用H37Rv-PPD和BCG-PPD分别在3 h、8 h、15 h及24 h四个时间点刺激分化成熟的THP-1细胞,再应用Hochest染色法,荧光镜下观察细胞的转归差异(凋亡及坏死情况),同时取上清用ELISA法测TNF-α、IL-1β及IL-10的浓度.结果 BCG-PPD刺激下细胞核以椭圆凋亡小体多见,而H37Rv-PPD刺激下细胞核则多呈坏死状,以坏死多见;BCG-PPD刺激下上清中TNF-α及IL-10的表达量低于H37Rv-PPD刺激组,但BCG-PPD刺激下IL-1β的表达量却高于后者.结论提示高毒力菌株衍生蛋白(H37Rv-PPD)引起THP-1坏死的原因可能与TNF-α的过度表达有关,而凋亡少见可能与IL-10抑制凋亡作用有关,而低毒力菌株衍生蛋白诱导凋亡与IL-1β有关.可能菌株毒力差异就存在于菌株的蛋白成分之中,且与上述几种细胞因子密切相关.
목적 본실험대불동독력결핵간균적연생단백(PPD)대인거서세포(THP-1)적영향급기여TNF-αt、IL-1 β급IL-10적차이성진행연구.방법 용H37Rv-PPD화BCG-PPD분별재3 h、8 h、15 h급24 h사개시간점자격분화성숙적THP-1세포,재응용Hochest염색법,형광경하관찰세포적전귀차이(조망급배사정황),동시취상청용ELISA법측TNF-α、IL-1β급IL-10적농도.결과 BCG-PPD자격하세포핵이타원조망소체다견,이H37Rv-PPD자격하세포핵칙다정배사상,이배사다견;BCG-PPD자격하상청중TNF-α급IL-10적표체량저우H37Rv-PPD자격조,단BCG-PPD자격하IL-1β적표체량각고우후자.결론 제시고독력균주연생단백(H37Rv-PPD)인기THP-1배사적원인가능여TNF-α적과도표체유관,이조망소견가능여IL-10억제조망작용유관,이저독력균주연생단백유도조망여IL-1β유관.가능균주독력차이취존재우균주적단백성분지중,차여상술궤충세포인자밀절상관.
Objective To study the different response in macrophages treated with different agoβ and IL-10 in Mycobacterium tuberculosisnists(H37Rv-PPD and BCG-PPD)related with Mycobacterium tuberculosis and the relationship with TNF-αt,IL-1β and IL-10.Methods Using H37Rv-PPD and BCG-PPD to stimulate THP-1 cell for 3h,8h,15h,24h respectively.Cells were ananlyzed by Hochest staining under fluorescence microscopy to assay cell death(apoptosis and necrosis).At each stimulating time,TNF-α,IL-1β and IL-10 were examined by ELISA.Results Under fluorescence microscopy,it could easily see oval apoptotic bodies of THP-1 stimulated by BCG-PPD.However ,the nucleus were often isolated and necrosis-like when cells were stimulated by H37Rv-PPD.In a word ,BCG-PPD tend to induce THP-1 cells to apoptosis,but H37Rv-PPD inclined to induce cells to undergo necrosis.In supernatant of cells stimulated by BCG-PPD,the expression of TNF-αand IL-10 were lower than the cells stimulated by H37Rv-PPD,but the expression of IL-1β was higher than the latter.Conclusion It indicated that the necrosis of cells stimulated by H37Rv-PPD was asossiated with the excessive expression of TNF-α and IL-10,and the apoptosis of cells induced by BCG-PPD was IL-1β related.Perhaps the mechanism of differences in virulence exist in protein of strain,and associated with cytokines IL-1β,TNF-α and IL-10.