CAJ | 학술논문

目的研究细菌脂多糖(LPS)耐受的THP-1细胞中沉默信息调节因子1(SIRTI)对IL-1β转录的调节作用.方法使用LPS耐受的人单核细胞THP-1模型,染色体免疫沉淀和real-time PCR定量研究1L-1β启动子区SIRT1结合情况和组蛋白H3lys9/H4lys16的乙酰化情况.结果在LPS耐受的THP-1细胞中,SIRTI对IL-1β启动子区的结合增加约5倍左右(P<0 05).同时伴随着组蛋白H3 lys9/H4 lys16乙酰化的低水平状态(与正常细胞相比P<0.05).SIRT1沉默使IL-1β的转录恢复到正常细胞的68%(P<0 05),同时伴随着组蛋白H3 lys9/H4lys 16乙酰化的增加(P<0.05).然而,正常细胞和耐受细胞p65 lys310乙酰化水平无明显差异.结论 SIRT1抑制LPS耐受的THP-1细胞中IL-113mRNA的转录,其作用与p65 lys310乙酰化无关,但是与IL-1β启动子区乙酰化有关.
목적 연구세균지다당(LPS)내수적THP-1세포중침묵신식조절인자1(SIRTI)대IL-1β전록적조절작용.방법 사용LPS내수적인단핵세포THP-1모형,염색체면역침정화real-time PCR정량연구1L-1β계동자구SIRT1결합정황화조단백H3lys9/H4lys16적을선화정황.결과 재LPS내수적THP-1세포중,SIRTI대IL-1β계동자구적결합증가약5배좌우(P<0 05).동시반수착조단백H3 lys9/H4 lys16을선화적저수평상태(여정상세포상비P<0.05).SIRT1침묵사IL-1β적전록회복도정상세포적68%(P<0 05),동시반수착조단백H3 lys9/H4lys 16을선화적증가(P<0.05).연이,정상세포화내수세포p65 lys310을선화수평무명현차이.결론 SIRT1억제LPS내수적THP-1세포중IL-113mRNA적전록,기작용여p65 lys310을선화무관,단시여IL-1β계동자구을선화유관.
Objective To explore the role of silent information regulation 2 homolog 1 (SIRTl) in the regulation of IL-lβ mRNA transcription in lipopolysaccharide(LPS) tolerant THP-1 cells. Methods THP-1 human promonocyte model of endotoxin tolerance that simulates the sepsis leukocyte phenotype was used. Chromatin immunoprecipitation assay (ChIP) and real-timePCR were applied to quantify the binding of SIRTl and histone H3 lys9/H4 lysl6 acetylation to IL-1β promoter. IL-1β mRNA transcription was studied after knocking down the SIRTl. Results Thebinding of SIRTl to IL-1β promoter increased about 5 times in tolerant THP-1 cells (P<0.05) , which was accompanied by the low level of histone H3 lys9/H4 lysl6 acetylation (P<0.05, compared with normal cells). Knocking-down of SIRTl increased the transcription of IL-1β mRNA up to the level of 68% of normal cells (P<0.05) ,which was accompanied by the increase of histone H3 lys9/H4 lysl6 acetylation (P<0.05). However,there was no significant difference of p65 lys310 acetylation between normal and tolerant cells. Conclusion SIRTl inhibited the IL-1 β mRNA transcription in tolerant THP-1 cells but had not related to p65 lys310 acetylation. However, it was related to IL-1 p promoter acetylation.