CAJ | 학술논문

目的通过观察小檗碱对脂多糖诱导的人单核细胞(THP-1)相关炎性反应因子表达的影响,探讨小檗碱的直接抗炎作用.方法小檗碱毒性作用分析:体外培养THP-1细胞,分为对照组和不同浓度小檗碱组(小檗碱5,10,20,50 μmol/L),分别孵育6h、24h、48 h,收集培养上清,应用乳酸脱氢酶(LDH)微量释放法检测不同浓度小檗碱对THP-1细胞的毒性作用.小檗碱对脂多糖诱导的THP-1细胞炎性因子的影响:分为对照组、脂多糖组(1μg/ml)、不同浓度小檗碱+脂多糖组(小檗碱5、10、20 μmol/L+1μg/ml脂多糖),分别孵育6h、24 h、48 h,收集培养上清,应用酶联免疫吸附试验(ELISA)检测白细胞介素(IL)-1β、IL-6、IL-8和肿瘤坏死因子(TNF)-α的水平.结果当小檗碱浓度＜20 μmol/L时,THP-1细胞经小檗碱干预6h、24 h、48 h之后,细胞存活率在90％以上,与对照组相比差异无统计学意义(P均＞0.05).小檗碱呈剂量依赖性的降低脂多糖诱导的THP-1细胞培养上清IL-1β、IL-6、IL-8和TNF-α水平.6h时,20 μmol/L小檗碱+脂多糖组IL-1β、IL-8、TNF-α水平与脂多糖组相比显著下降(P均＜0.05);24 h时,20 μmol/L小檗碱+脂多糖组IL-8和TNF-α水平与脂多糖组相比均显著下降(P均＜0.05);48 h时,5μmol/L小檗碱+脂多糖组TNF-α水平显著低于脂多糖组(F=92.625,P＜0.05);10 μmol/L小檗碱+脂多糖组IL-1β、IL-6和TNF-α水平亦显著低于脂多糖组(P均＜0.05);20 μmol/L小檗碱+脂多糖组IL-1β、IL-6、IL-8和TNF-α水平亦显著低于脂多糖组(P均＜0.05).结论小檗碱可显著降低脂多糖诱导的THP-1细胞分泌炎性反应因子,且具有一定的剂量-效应关系.
목적 통과관찰소벽감대지다당유도적인단핵세포(THP-1)상관염성반응인자표체적영향,탐토소벽감적직접항염작용.방법 소벽감독성작용분석:체외배양THP-1세포,분위대조조화불동농도소벽감조(소벽감5,10,20,50 μmol/L),분별부육6h、24h、48 h,수집배양상청,응용유산탈경매(LDH)미량석방법검측불동농도소벽감대THP-1세포적독성작용.소벽감대지다당유도적THP-1세포염성인자적영향:분위대조조、지다당조(1μg/ml)、불동농도소벽감+지다당조(소벽감5、10、20 μmol/L+1μg/ml지다당),분별부육6h、24 h、48 h,수집배양상청,응용매련면역흡부시험(ELISA)검측백세포개소(IL)-1β、IL-6、IL-8화종류배사인자(TNF)-α적수평.결과 당소벽감농도＜20 μmol/L시,THP-1세포경소벽감간예6h、24 h、48 h지후,세포존활솔재90％이상,여대조조상비차이무통계학의의(P균＞0.05).소벽감정제량의뢰성적강저지다당유도적THP-1세포배양상청IL-1β、IL-6、IL-8화TNF-α수평.6h시,20 μmol/L소벽감+지다당조IL-1β、IL-8、TNF-α수평여지다당조상비현저하강(P균＜0.05);24 h시,20 μmol/L소벽감+지다당조IL-8화TNF-α수평여지다당조상비균현저하강(P균＜0.05);48 h시,5μmol/L소벽감+지다당조TNF-α수평현저저우지다당조(F=92.625,P＜0.05);10 μmol/L소벽감+지다당조IL-1β、IL-6화TNF-α수평역현저저우지다당조(P균＜0.05);20 μmol/L소벽감+지다당조IL-1β、IL-6、IL-8화TNF-α수평역현저저우지다당조(P균＜0.05).결론 소벽감가현저강저지다당유도적THP-1세포분비염성반응인자,차구유일정적제량-효응관계.
Objective To observe the effects of berberine (BBR) on expression of inflammatory cytokines in THP-1 cells induced by lipopolysaccharide (LPS),and investigate the anti-inflammatory effects of BBR.Methods For analysing the toxicity of BBR on THP-1 cells,THP-1 cells were divided into control group,and different concentrations of BBR groups (BBR 5,10,20,50 μmol/L).After incubation for 6,24 and 48 hours,lactate dehydrogenase (LDH) released from THP-1 cells was used to assay the cytotoxicity of BBR.For analysis of the effects of BBR on inflammatory cytokines induced by LPS in THP-1 cells,THP-1 cells were divided into control group,LPS group (1 μg/mL LPS),and different concentrations of BBR with LPS groups (BBR 5,10 and 20 μmol/L + 1 μg/mL LPS).After 6,24 or 48 hours of incubation,the concentrations of inter4eukin (IL)-1 β,IL-6,IL-8 and tumor necrosis factor(TNF)-α in the culture medium were measured by enzyme-linked immunosorbent assay (ELISA).Results The survival rates of THP-1 cells were all over 90％after treated with BBR lower than 20 μmol/L for 6,24 and 48 hours.BBR decreased the release of IL-1β,IL-6,IL-8 and TNF-α from THP-1 cells in a dose-dependent manner.After 6 hours,20 μmol/L of BBR decreased the secretion of IL-1 β,IL-8 and TNF-α significantly compared with LPS group (P ＜ 0.05).After 24 hours,the secretion of IL-8 and TNF-α was decreased significantly in 20 μmol/L of BBR + LPS group compared with those in LPS group (P ＜ 0.05).After 48 hours,the secretion of TNF-α was decreased signifi-cantly (F=92.625,P ＜ 0.05) in 5 μmol/L BBR + LPS group,the secrection of IL-1β、IL-6 and TNF-α were decreased (all P ＜ 0.05) in 10 μmol/L BBR + LPS group,and the secretion of IL-1β,IL-6 and TNF-α were decreased in 20 μmol/L BBR + LPS group compared with those in LPS group (all P ＜ 0.05).Conclusion BBR can decrease the secretion of inflammatory cytokines in THP-1 cells induced by LPS in a dose-dependent manner.