摘要
目的:研究右美托咪定(Dex)通过α2肾上腺能受体对脂多糖(LPS)诱导的库普弗细胞(KCs)炎性反应及血红素氧合酶(HO-1)表达的影响。方法:将来源于SD小鼠的KCs细胞接种在30孔板上,采用随机数表法将其分为空白组、对照组、Dex-L组、Dex-M组和Dex-H组,空白组加入细胞培养液培养24h,对照组加入μg/ml的LPS培养24h,Dex-L组、Dex-M组和Dex-H组与对照组进行同样处理后,再分别加入浓度为5 ng/ml、10ng/ml及20ng/ml的右美托咪定培养1h。每组设置6个孔,在孵育6h和12h时收集细胞上清液,采用酶联免疫吸附测定(ELISA)法测定每组细胞的肿瘤坏死因子α(TNF-α)、白细胞介素-6(IL-6)及晚期炎症介质高迁移率族蛋白(HMGB-1)的浓度水平,采用Western blot法测量每组细胞的HO-1蛋白表达水平。培育24h时采用细胞计数试剂(CCK-8)法计算每组细胞的存活率。结果:培养6h和121h时,Dex-L组、Dex-M组和Dex-H组的TNF-α、IL-6及HMGB-1浓度比空白组明显升高,差异有统计学意义(t_(Dex-L组)=3.891,t=3.179,t=3.588;t_(Dex-M组)=3.663,t=3.765,t=3.501;t_(Dex-H组)-4.134,t=4.333,t=4.285;P<0.05);3组与对照组比较3种炎性因子明显降低(t_(Dex-L组)=3.451,t=3.682,t=3.509;t_(Dex-)M组=3.919,t=3.383,t=3.286;t_(Dex-H组)=4.455,t=4.136,t=4.481,P<0.05);细胞培养6h_后3组炎性因子TNF-α、IL-6和HMGB-1浓度水平与对照组比较,差异有统计学意义(F=6.346,F=12.973,F=8.325,P<0.05)。3组细胞培养6h和12h时HO-1水平比较,差异有统计学意义(F=4.827,F=6.124;P<0.05)。培养24h后,各组细胞存活率无显著性差异。结论:LPS能诱导KCs细胞释放炎性因子,而Dex可能通过调节α2肾上腺能受体上调HO-1水平抑制该过程从而发挥抗炎作用。
Objective:To study the effects of dexmedetomidine(Dex) on inflammatory response and HO-1 expression of lipopolysaccharide(LPS)-induced KCs through α2 adrenergic receptors.Methods:The KCs cells from SD mice were inoculated on 30-well plates,and they were randomly divided into 5 groups(the blank group,control group,Dex-L group,Dex-M group and Dex-H group) according to random number table.The cell culture solution was added in blank group for culture of 24 h,and then Dex of 5 ng/mL,10 ng/mL and 20 ng/mL were added respectively in Dex-L group,Dex-M group and Dex-H group for culture of 1 h.Each group included 6 wells,and the supernatant of cell in each well was collected at the 6 th h and 12 th h post culture.And the enzyme-linked immunosorbent assay(ELISA) was adopted to detect the concentrations of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6) and high mobility group box-1(HMGB-1) of inflammatory mediator at advanced stage.And Western blot was used to detect the expression level of H0-1 protein in cells of each group.The cell counting kit-8(CCK-8) of cell count reagent was adopted to calculate the survival rate of cells in each group after cells were cultured for 24 h.Results:At the 6 th h and 12 th h of culture,the concentrations of TNF-α,IL-6 and HMGB-1 of Dex-L group,Dex-M group and Dex-H group were significantly higher than those of blank group(tDex-L group=3.891,t=3.179,t=3.588,tDex-M group=3.663,t=3.765,t=3.501,tDex-H group=4.134,t=4.333,t=4.285,P<0.05),respectively.And the expression level of inflammatory factor of three groups were significantly lower than those of control group(tDex-L group=3.451,t=3.682,t=3.509,tDex-M group=3.919,t=3.383,t=3.286,tDex-H group=4.455,t=4.136,t=4.481,P<0.05,respectively.After cells were cultured for 6 h,the differences of the concentrations of TNF-α,IL-6 and HMGB-1 among the three groups and control group were significantly(F=6.346,F=12.973,F=8.325,P<0.05).And the differences of expression level of H0-1 protein at the 6 th h and 12 th h of cell culture among three gr
作者
康文越
邢丹丹
俞晓东
付强
林慧
KANG Wen-yue;XING Dan-dan;YU Xiao-dong(Department of Anesthesiology,Hainan General Hospital,Haikou 570300,China)
出处
《中国医学装备》
2021年第1期153-157,共5页
China Medical Equipment
基金
海南省卫生计生行业科研项目(1601320212A2001)“盐酸右美托咪定对LPS诱导的Kupffer细胞氧化应激和炎性反应的影响”。
