全反式维甲酸对高糖环境下HK-2细胞炎症因子表达的影响

Effect of All-trans Retinoic Acid on High Glucose Induced-Expression of Inflammatory Factors in HK-2 Cells

目的探讨全反式维甲酸(ATRA)对高糖环境下正常人肾小管上皮细胞(HK-2细胞)炎症因子表达的影响及其可能机制。方法将体外培养的HK-2细胞随机分为7组:空白对照组,高糖组(D-葡萄糖30 mmol·L^(-1)),高渗组(甘露醇24.5 mmol·L^(-1)),ATRA干预组[包括高糖+低浓度ATRA组(ATRA 10-7mol·L^(-1))、高糖+中浓度ATRA组(ATRA 10-6mol·L^(-1))、高糖+高浓度ATRA组(ATRA 10-5mol·L^(-1))],Rho激酶抑制药(Y27632)干预组[高糖+Y27632(Y27632为30μmol·L^(-1))],均干预48 h。逆转录-聚合酶链反应(RT-PCR)法检测细胞Rho A mRNA、ROCK1 mRNA表达水平,酶联免疫吸附法(ELISA)检测HK-2细胞白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)变化。结果 RT-PCR结果显示,空白对照组与高渗组Rho A mRNA、ROCK1 mRNA表达差异无统计学意义(P>0.05);高糖组Rho A mRNA、ROCK1 mRNA表达较空白对照组、高渗组显著升高(P<0.05);ATRA各干预组Rho A mRNA、ROCK1 mRNA表达较高糖组显著减少(P<0.05),且呈现ATRA浓度依赖性;Y27632干预组Rho A mRNA表达与高糖组差异无统计学意义,ROCK1mRNA表达显著减少(P<0.05)。Person直线相关分析显示,高糖组及ATRA干预组Rho A mRNA与ROCK1 mRNA表达呈正相关。ELISA检测结果示空白对照组与高渗组表达IL-6、TNF-α无明显差异(P>0.05);高糖组表达IL-6、TNF-α较空白对照组明显升高(P<0.05);ATRA或Y27632干预后,IL-6、TNF-α表达明显减少(P<0.05)。结论 ATRA可抑制高糖环境下HK-2细胞IL-6、TNF-α表达,其机制可能与抑制Rho A/ROCK信号通路有关。

Objective To explore the effect of all-trans retinoic acid（ATRA） on inflammatory cytokines expression in human renal tubular epithelial cells under high glucose condition and its possible mechanisms. Methods HK-2 cells were randomly divided into seven groups： blank control group, high glucose group, hypertonie group, ATRA intervention groups and Rho kinase inhibitor （Y27632） intervention group. All groups were treated for 48 hours. Reverse transcription polymerase chain reaction （RT-PCR） was used to evaluate RhoA and ROCK1 mRNA levels.ELISA was used to detect interleukin-6 （IL-6） and tumor necrosis factor-ct （TNF- α） expression in HK-2 cells. Results RT-PCR showed that mRNA expressions of RhoA and ROCK1 were not different between the hypertonic group and the blank group （P〉0.05）, but were significantly increased in the high glucose group （P〈0.05）.In the ATRA treatment group, RhoA and ROCK1 mRNA expressions were significantly decreased compared with the high glucose group in a concentration-dependent manner. RhoA mRNA expressions showed no difference between the Y27632 intervention group and the high glucose group, but ROCK1 mRNA expression was significantly reduced （P〈 0.05）. By Pearson correlation analysis, we found that there was a positive correlation between RhoA and ROCK1 mRNA expressions in the high glucose group and the ATRA treatment group.ELISA showed that the IL-6 and TNF-α protein levels were not different between the control and the hypertonic group （P〉0.05）, but were significantly increased in the high glucose group and decreased in the ATRA or Y27632 treatment groups （P〈0.05）. Conclusion ATRA decreases IL-6 and TNF-α protein levels in HK-2 cells induced by high glucose. The underlying mechanism may be associated with the RhoA/ROCK signaling pathway.