基于LncRNA H19-P50 NF-κB信号通路探讨肢体缺血-再灌注损伤机制及桃红四物液的干预作用

Based on the LncRNA H19-P50 NF-κB signal pathway to explore the mechanism of limb ischemia-reperfusion injury and the intervention effect of Taohong Siwu Liquid

目的观察桃红四物液对肢体缺血-再灌注损伤中LncRNA H19、NF-κB P50的表达影响,探讨肢体缺血-再灌注损伤的机制。方法51只2月龄雄性SD大鼠,按随机数表随机分为正常对照组(3只)、模型对照组(12只)、单纯H19阻断剂组(12只)、H19阻断剂组+桃红四物液组(12只)、桃红四物液组(12只),各组动物分别于再灌注0h、2h、4h、8h等不同时间点取相应的标本,用于Western-blot检测NF-κB P50蛋白的表达、RT-PCR检测H19、NF-κB P50的mRNA的表达。结果较正常对照组相比,模型对照组、单纯H19阻断剂组、H19阻断剂组+桃红四物液组、桃红四物液组的H19、NF-κB P50的mRNA表达均升高,NF-κB P50蛋白的表达亦增高,差异有统计学意义(P<0.05);较模型对照组相比,单纯H19阻断剂组、H19阻断剂组+桃红四物液组、桃红四物液组的H19、NF-κB P50的mRNA表达均降低,NF-κB P50蛋白的表达亦降低,差异有统计学意义(P<0.05);较H19阻断剂组+桃红四物液组相比,单纯H19阻断剂组和桃红四物液组的H19、NF-κB P50的mRNA表达均降低,NF-κB P50蛋白的表达亦降低,差异有统计学意义(P<0.05)。结论:LncRNA H19是LIRI中的关键分子,NF-κB P50是LIRI中的关键蛋白,桃结论红四物液可以通过调控LncRNA H19影响NF-κB P50的表达,从而对肢体缺血-再灌注损伤产生作用。

Objective To observe the effect of Taohong Siwu Liquid on the expression of LncRNA H19 and NF-κB P50 in limb ischemia-reperfusion injury, and to explore the mechanism of limb ischemia-reperfusion injury.Methods Fifty-one 2-month-old male SD rats were randomly divided into normal control group(3 rats), model control group(12 rats), pure H19 blocker group(12 rats), and H19 blocker according to the random number table. Group + Taohong Siwuye group(12 animals), Taohong Siwuye group(12 animals), the animals in each group were reperfused at 0 h, 2 h, 4 h, 8 h and other time points to take corresponding specimens for Western-blot detection of NF-κB P50 protein expression, RT-PCR detection of H19, NF-κB P50 mRNA expression.Results Compared with the normal group, the mRNA expressions of H19 and NF-κB P50 in the model control group, pure H19 blocker group, H19 blocker group + Taohong Siwuye group, and Taohong Siwuye group increased. The expression of NF-κB P50 protein also increased, and the difference was statistically significant(P<0.05);compared with the model control group, pure H19 blocker group, H19 blocker group + Taohong Siwu liquid group, Taohong Siwu group The mRNA expression of H19 and NF-κB P50 in the liquid group decreased, and the expression of NF-κB P50 protein also decreased, the difference was statistically significant(P<0.05);compared with H19 blocker group + Taohong Siwu liquid group, The mRNA expression of H19 and NF-κB P50 in the pure H19 blocker group and the Taohong Siwuye group decreased, and the expression of NF-κB P50 protein also decreased, the difference was statistically significant(P<0.05).Conclusion LncRNA H19 is the key molecule in LIRI, and NF-κB P50 is the key protein in LIRI. Taohong Siwu Liquid can affect the expression of NF-κB P50 by regulating LncRNA H19, thereby exerting effects on limb ischemia-reperfusion injury.