乌司他丁通过miR-146a调节TLR4/NF-κB信号通路减轻失血性休克大鼠肾炎性损伤研究

王振杰; 陈硬; 宋琦; 徐志鹏; 纪忠; 邱兆磊; 郑传明; 李磊; 程峰

doi:10.13898/j.cnki.issn.1000-2200.2020.03.002

乌司他丁通过miR-146a调节TLR4/NF-κB信号通路减轻失血性休克大鼠肾炎性损伤研究

Study on ulinastatin alleviating the nephritis injury in hemorrhagic shock rats by regulating the TLR4/NF-κB signaling pathway via miR-146a

摘要

摘要:
目的探讨乌司他丁能否在醋酸钠林格液的基础上影响miR-146a及TLR4/NF-κB信号通路以进一步削弱失血性休克大鼠的肾组织炎性损伤。
方法24只健康SD大鼠建立失血性休克模型，分为休克未复苏组（SR组）、醋酸钠林格液复苏组（AR组）和乌司他丁联用醋酸钠林格液复苏组（UR组），每组各8只。应用实时定量PCR技术检测各组大鼠肾组织中miR-146a和促炎因子肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1、IL-6及抑炎因子IL-4、IL-10的mRNA表达量，并通过Western blotting检测TLR4、NF-κB的相对蛋白表达量。光镜下观察各组大鼠肾组织在病理学方面的改变。
结果与SR组比较，AR组和UR组肾组织中miR-146a的mRNA表达水平均有明显升高（P < 0.01）；而与AR组相比，UR组中miR-146a的mRNA表达仍有升高（P < 0.01）。与SR组相比，AR组和UR组中TNF-α的mRNA表达水平均有明显降低（P < 0.01），而与AR组相比，UR组中TNF-α的mRNA表达仍有明显降低（P < 0.01）。与SR组比较，AR组和UR组中IL-1和IL-6的mRNA表达均有明显降低（P < 0.01），而UR组与AR组差异无统计学意义（P>0.05）。与SR组和AR组相比，UR组中IL-4和IL-10的mRNA表达均明显升高（P < 0.01）；而SR组和AR组相比，差异无统计学意义（P>0.05）。与SR组相比，AR组和UR组的TLR4和NF-κB蛋白表达水平均有明显降低（P < 0.01），而与AR组相比，UR组中TLR4和NF-κB的表达仍有明显降低（P < 0.01）。光镜下可见，与SR组相比，AR组和UR组肾组织的病理损伤程度均有明显减轻，但UR组较AR组减轻程度更为明显。
结论乌司他丁可能在醋酸钠林格液减轻创伤失血性休克大鼠肾组织炎性损伤的基础上进一步发挥保护作用，其机制可能与上调miR-146a的表达，抑制TLR4/NF-κB信号通路及调控炎症因子表达有关。

Abstract:
ObjectiveTo investigate the effects of ulinastatin on the miR-146a and TLR4/NF-κB signaling pathways based on sodium acetate Ringer's solution to further weaken the inflammatory damage of renal tissue in hemorrhagic shock rats.
MethodsThe hemorrhagic shock model in 24 healthy SD rats were established, and divide into the shocked non-resuscitation group(SR group, n=8), sodium acetate Ringer's solution resuscitation group(AR, n=8) and ulinastatin combined with sodium acetate Ringer's solution resuscitation group(UR, n=8).The mRNA expression levels of miR-146a and inflammatory factors(TNF-α, IL-1, IL-4, IL-6 and IL-10) in the kidney tissue in three group were detected using the real-time quantitative PCR technology, the expression levels of TLR4 and NF-κB protein were detected using Western blot, and the pathological changes of the kidney tissues of rats in each group were observed under light microscope.
ResultsCompared with the SR group, the expression levels of miR-146a mRNA in the renal tissues of the AR group and UR group significantly increased(P < 0.01), and compared with the AR group, the expression level of miR-146a mRNA in the UR group increased(P < 0.01).Compared with the SR group, the expression levels of TNF-α mRNA in the AR group and UR group significantly decreased(P < 0.01), while the expression level of TNF-α mRNA in the UR group still significantly decreased compared with the AR group(P < 0.01).Compared with the SR group, the expression levels of IL-1 and IL-6 mRNA in the AR group and UR group significantly decreased(P < 0.01), and there was no statistical significance in the expression levels of IL-1 and IL-6 mRNA between the UR group and AR group(P>0.05).Compared with the SR group and AR group, the expression levels of IL-4 and IL-10 mRNA in the UR group significantly increased(P < 0.01), and the differences of those between SR group and AR group were not statistically significant(P>0.05).Compared with the SR group, the expression levels of TLR4 and NF-κB protein in the AR group and UR group significantly decreased(P < 0.01).Compared with the AR group, the expression levels of TLR4 and NF-κB protein in the UR group still significantly decreased(P < 0.01).Under the light microscope, compared with the SR group, the pathological damage degree of kidney tissue in the AR group and UR group significantly alleviated, and the relieving degree of kidney tissue injury in UR group was more significant than that in AR group.
ConclusionsUlinastatin may play a further protective role on the basis of sodium acetate Ringer's solution alleviating the inflammatory injury of renal tissue in rats with traumatic hemorrhagic shock, and the mechanism of which may be related to the up-regulation of miR-146a, inhibition of the TLR4/NF-κB signaling pathway and regulation of the expression of inflammatory factors.

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