CAJ | 학술논문

目的观察脂多糖(LPS)对BV-2细胞形状变化及白介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)分泌情况的影响,探讨沉默信息调控因子1(SIRT1)在LPS诱导BV-2细胞释放前炎症因子过程中的作用. 方法 BV-2细胞分为调零组、对照组及LPS组,调零组不含细胞,只加培养基;对照组常规培养；LPS组分别加入不同浓度的LPS; MTT法检测各组存活率.另取BV-2细胞分为调零组、对照组、DOSO药物载体组及白藜芦醇组、Sirtinol组,调零组、对照组处理方法同前,DOSO药物载体组加入体积分数为0.3％的DMSO;白藜芦醇组加入10、25、50、100 μmol/L白藜芦醇;Sirtinol组加入1、2.5、5、10、25、50 μmol/L Sirtinl; MTT法检测各组细胞存活率.根据实验结果,选取适当浓度的LPS、白藜芦醇及Sirtinol,将BV-2细胞分为对照组、LPS组、白藜芦醇+LPS组、Sirtinol +LPS组,分别用ELISA方法检测各组细胞相应处理12、24 h后上清液中的IL-6和TNF-α的含量,Western blotting检测SIRT1在上述各组细胞加药处理24 h后的表达水平. 结果与对照组相比,LPS组BV-2细胞数目增多,胞体肿胀,突起变短、增多.随着LPS浓度的增加,BV-2细胞存活率明显增加,差异有统计学意义(P＜0.05).与对照组比较,LPS组BV-2细胞分泌的IL-6、TNF-α增多,S IRT1的表达量下降,差异有统计学意义(P＜0.05).与LPS组比较,经白藜芦醇处理后,细胞内SIRT1的表达量上升,而IL-6 、TNF-α的分泌水平下降,差异有统计学意义(P＜0.05)；相反,细胞经Sirtinol处理后,胞内SIRT1的表达量下降,IL-6、TNF-α的分泌水平上升,差异有统计学意义(P＜0.05). 结论 LPS可显著改变BV-2细胞形状并诱导细胞前炎症因子的表达,干预SIRT1的表达量对上述现象有明显的调节作用.
목적 관찰지다당(LPS)대BV-2세포형상변화급백개소-6(IL-6)화종류배사인자-α(TNF-α)분비정황적영향,탐토침묵신식조공인자1(SIRT1)재LPS유도BV-2세포석방전염증인자과정중적작용. 방법 BV-2세포분위조령조、대조조급LPS조,조령조불함세포,지가배양기;대조조상규배양；LPS조분별가입불동농도적LPS; MTT법검측각조존활솔.령취BV-2세포분위조령조、대조조、DOSO약물재체조급백려호순조、Sirtinol조,조령조、대조조처리방법동전,DOSO약물재체조가입체적분수위0.3％적DMSO;백려호순조가입10、25、50、100 μmol/L백려호순;Sirtinol조가입1、2.5、5、10、25、50 μmol/L Sirtinl; MTT법검측각조세포존활솔.근거실험결과,선취괄당농도적LPS、백려호순급Sirtinol,장BV-2세포분위대조조、LPS조、백려호순+LPS조、Sirtinol +LPS조,분별용ELISA방법검측각조세포상응처리12、24 h후상청액중적IL-6화TNF-α적함량,Western blotting검측SIRT1재상술각조세포가약처리24 h후적표체수평. 결과 여대조조상비,LPS조BV-2세포수목증다,포체종창,돌기변단、증다.수착LPS농도적증가,BV-2세포존활솔명현증가,차이유통계학의의(P＜0.05).여대조조비교,LPS조BV-2세포분비적IL-6、TNF-α증다,S IRT1적표체량하강,차이유통계학의의(P＜0.05).여LPS조비교,경백려호순처리후,세포내SIRT1적표체량상승,이IL-6 、TNF-α적분비수평하강,차이유통계학의의(P＜0.05)；상반,세포경Sirtinol처리후,포내SIRT1적표체량하강,IL-6、TNF-α적분비수평상승,차이유통계학의의(P＜0.05). 결론 LPS가현저개변BV-2세포형상병유도세포전염증인자적표체,간예SIRT1적표체량대상술현상유명현적조절작용.
Objective To observe the effect oflipopolysaccharide (LPS) on the cell form of BV-2 cells and the expressions of interleukin-6 (IL-6) and tumor necrosis factor-a (TNF-a) so as to detect the role of silence information regulator 1 (SIRT1) in regulation of LPS-induced proinflammatory cytokines production in activated BV-2 cells.Methods BV-2 cells were divided into control group (normal culture medium) and treatment groups; BV-2 cells in the treatment groups were subdivided into LPS treatment groups,Resveratrol+LPS treatment groups and Sirtinol+LPS treatment groups (cultured with different concentrations of LPS,SIRT1 activator Resveratrol or SIRT1 inhibitor Sirtinol,respectively).MTT assay was employed to identify the cell survival after the inducement.Based on the above MTT results,the cells were then grouped into the control group,LPS treatment group,Resveratrol+LPS treatment group and Sirtinol+LPS treatment group having suitable concentrations of LPS,Resveratrol and Sirtinol; then,the levels of IL-6 and TNF-a were measured with enzyme-linked immuno sorbent assay (ELISA) at 12 and 24 h after the inducement; and the expression of SIRT1 at 24 hafter the inducement was detected by Western blotting.Results As compared with those in the control group,the BV-2 cells in the LPS treatment group had increased cell number,hypertrophic cell body,and shorten cell processes.The cell survival rate increased with increased concentrations of LPS.As compared with those in the control group,the levels of IL-6 and TNF-a in LPS treatment group increased and level of SIRT 1 decreased with significant differences (P＜0.05).Significantly increased levels of IL-6and TNF-a and obviously decreased expression of SIRT1 in the Resveratrol+LPS treatment group were noted as compared with those in the LPS treatment group (P＜0.05); conversely,significantly decreased levels of IL-6 and TNF-a and obviously increased expression of SIRT1 in the Sirtinol+LPS treatment group were noted as compared with those in the LPS treatment group (P＜0.05).Conclusion LPS can change the morphology of BV-2 cells,and induce the levels ofproinflammatory cytokines; impairment of SIRT1 may contribute to such progress obviously.