右美托咪定预处理对肾小管上皮细胞缺氧/复氧损伤的抗炎作用

Anti-inflammatory effect of Dexmedetomidine pretreatment on hypoxia/reoxygenation-induced injury in human renal tubular epithelial cells

目的探讨右美托咪定预处理对肾小管上皮细胞缺氧/复氧损伤的抗炎作用。方法体外培养人肾小管上皮细胞(HK-2),以1. 5×104/ml的密度接种于96孔培养板(每孔200μl)或以2×105/ml密度接种于6孔培养板(每孔2 ml)。采用随机数字表法分为4组(n=24):正常对照组(CON组)、右美托咪定组(DEX组)、缺氧复氧组(H/R组)、缺氧复氧+右美托咪定组(H/R+DEX组)。DEX组和H/R+DEX组加入0. 1 nmol/L的右美托咪定孵育2 h,随后CON组和DEX组细胞置于常氧恒温细胞培养箱28 h,H/R组和H/R+DEX组细胞缺氧24 h复氧4 h。MTT检测各组细胞活力,酶联免疫吸附法(ELISA)检测细胞上清和细胞内肿瘤坏死因子α(TNF-α)、白介素-1β(IL-1β)和白介素-10(IL-10)的水平。结果与CON组相比,H/R组和H/R+DEX组细胞生长状态转坏,细胞活力明显下降,TNF-α、IL-1β浓度升高,IL-10浓度降低(P <0. 05); DEX组上述指标差异无统计学意义(P> 0. 05);与H/R组相比,H/R+DEX组细胞生长状态转好,细胞活力升高,TNF-α、IL-1β浓度降低,IL-10浓度升高(P <0. 05)。结论右美托咪定预处理可以抑制缺氧复氧引起的肾小管上皮细胞的损伤,并减轻其炎症反应。

Objective:To investigate the anti-inflammatory effect of Dexmedetomidine pretreatment on hypoxia/reoxygenation-induced injury in human renal tubular epithelial cells.Methods:Cultured human renal tubular epithelial cells(HK-2)were seeded in 96-well plates with a density of 1.5×10 4/ml(200μl/well)or in 6-well plates(2 ml/well)with a density of 2×10 5/ml,and randomly divided into four groups:group control(CON),group Dexmedetomidine(DEX),group hypoxia/reoxygenation(H/R),group hypoxia/reoxygenation plus Dexmedetomidine(H/R+DEX).In groups DEX and H/R+DEX,Dexmedetomidine 0.1 nmol/L was added to the culture medium and the cells were incubated for 2 h.Then groups CON and DEX were exposed to regular incubator and incubated for 28h,groups H/R and H/R+DEX were incubated in an anaerobic chamber for 24h and then returned to a regular incubator and incubated for 4 h.Cell growth conditions were observed under inverted microscope,the cell viability were measured by MTT assay.ELISA method was measured the level of TNF-α、IL-1β、IL-8.Results:Compared with group CON,the cell condition was worsened and survival rate was significantly decreased,The concentration of TNF-α、IL-1βwere significantly increased,whereas IL-10 was decreased significantly in group H/R and group H/R+DEX(P<0.05),No significant difference was found in group DEX(P>0.05);Compared with group H/R,the cell condition was improved and survival rate was significantly increased,The concentration of TNF-α、IL-1βwere significantly decreased,whereas IL-10 was increased significantly in group H/R+DEX(P<0.05).Conclusion:Dexmedetomidine pretreatment can effectively attenuate hypoxia/reoxygenation-induced injury to human renal tubular epithelial cells,and ameliorates inflammatory reactions.