CAJ | 학술논문

目的评价盐酸戊乙奎醚对脂多糖(LPS)诱导小鼠巨噬细胞高迁移率族蛋白B1(HMGB1)合成和释放的影响.方法将小鼠RAW264.7细胞接种于培养皿中,常规培养24h,采用随机数字表法,将其分为5组(n=6).正常对照组(C组):常规培养;LPS组:加入LPS终浓度为500ng/ml;P1组:同时加入LPS和盐酸戊乙奎醚,终浓度分别为500 ng/ml、0.5 μg/ml;P2组:同时加入LPS和盐酸戊乙奎醚,终浓度分别为500 ng/ml、2μg/ml;P3组:同时加入LPS和盐酸戊乙奎醚终浓度分别为500 ng/ml、5 μg/ml;孵育24 h,收集细胞及上清液,采用CCK-8细胞计数法测定细胞增殖水平;RT-PCR法测定细胞HMGB1 mRNA表达;Western blot法测定细胞NF-κBp65表达;ELISA法测定上清液HMGB1浓度.结果与C组比较,LPS组、P1组、P2组及P3组细胞HMGB1 mRNA和NF-κBp65的表达上调,上清液HMGB1浓度升高(P＜0.05);与LPS组比较,P2组及P3组细胞HMGB1 mRNA和NF-κBp65的表达下调,上清液HMGB1浓度降低(P＜0.05);与P1组比较,P2组及P3组细胞HMGB1 mRNA和NF-κBp65的表达下调,上清液HMGB1浓度降低(P＜0.05);与P2组比较,P3组上述各指标差异无统计学意义(P＞0.05).5组细胞增殖水平差异无统计学意义(P＞0.05).结论盐酸戊乙奎醚可通过抑制NF-κB活化,降低LPS诱导小鼠巨噬细胞HMGB1的合成和释放.
목적 평개염산무을규미대지다당(LPS)유도소서거서세포고천이솔족단백B1(HMGB1)합성화석방적영향.방법 장소서RAW264.7세포접충우배양명중,상규배양24h,채용수궤수자표법,장기분위5조(n=6).정상대조조(C조):상규배양;LPS조:가입LPS종농도위500ng/ml;P1조:동시가입LPS화염산무을규미,종농도분별위500 ng/ml、0.5 μg/ml;P2조:동시가입LPS화염산무을규미,종농도분별위500 ng/ml、2μg/ml;P3조:동시가입LPS화염산무을규미종농도분별위500 ng/ml、5 μg/ml;부육24 h,수집세포급상청액,채용CCK-8세포계수법측정세포증식수평;RT-PCR법측정세포HMGB1 mRNA표체;Western blot법측정세포NF-κBp65표체;ELISA법측정상청액HMGB1농도.결과 여C조비교,LPS조、P1조、P2조급P3조세포HMGB1 mRNA화NF-κBp65적표체상조,상청액HMGB1농도승고(P＜0.05);여LPS조비교,P2조급P3조세포HMGB1 mRNA화NF-κBp65적표체하조,상청액HMGB1농도강저(P＜0.05);여P1조비교,P2조급P3조세포HMGB1 mRNA화NF-κBp65적표체하조,상청액HMGB1농도강저(P＜0.05);여P2조비교,P3조상술각지표차이무통계학의의(P＞0.05).5조세포증식수평차이무통계학의의(P＞0.05).결론 염산무을규미가통과억제NF-κB활화,강저LPS유도소서거서세포HMGB1적합성화석방.
Objective To evaluate the effects of penehyclidine hydrochloride on synthesis and release of high mobility group protein box 1 (HMGB1) in macrophages induced by lipopolysaccharide (LPS) in mice.Methods RAW264.7 cells obtained from mice were seeded in the culture dishes.After being cultured for 24 h,the cells were randomly divided into 5 groups (n =6 each) using a random number table.The cells were incubated routinely in group C.The cells were incubated in the presence of LPS 500 ng/ml (group LPS),LPS 500 ng/ml + penehyclidine hydrochloride 0.5 μg/ml (group P1),LPS 500 ng/ml + penehyclidine hydrochloride 2 μg/ml (group P2),or LPS 500 ng/ml + penehyclidine hydrochloride 5 μg/ml (group P3).All the cells were incubated for 24 h.The cells and supernatant were collected.The proliferation of the cells was measured by CCK-8 assay,HMGB1 mRNA expression in the cells was detected by RT-PCR,NF-κBp65 protein expression in the cells was detected by Western blot and the concentration of HMGB1 in the supernatant was detected by ELISA.Results Compared with group C,the expression of HMGB1 mRNA and NF-κBp65 protein in the cells was significantly upregulated,and the concentration of HMGB1 in the supernatant was increased in LPS,P1,P2 and P3 groups (P ＜ 0.05).Compared with group LPS,the expression of HMGB1 mRNA and NF-κBp65 protein in the cells was significantly down-regulated,and the concentration of HMGB1 in the supernatant was decreased in P2 and P3 groups (P ＜ 0.05).Compared with group P1,the expression of HMGB1 mRNA and NF-κBp65 protein in the cells was significantly down-regulated,and the concentration of HMGB1 in the supernatant was decreased in P2 and P3 groups (P ＜ 0.05).There was no significant difference in the indicators mentioned above between group P2 and group P3 (P ＞ 0.05).There was no significant difference in the proliferation of the cells among the 5 groups (P ＞ 0.05).Conclusion Penehyclidine hydrochloride can reduce the synthesis and release of HMGB1 in macrophages induced by LPS through inhibiting NF-κB activation in mice.