血必净注射液通过调控TLR4/MyD88/NF-κB信号通路改善脓毒症诱导大鼠急性肾损伤作用机制研究

Mechanism of Xuebijing Injection in ameliorating sepsis-induced acute kidney injury by regulating TLR4/MyD88/NF-κB signaling pathway in rats

关键词:  血必净注射液  脓毒症  急性肾损伤  TLR4/MyD88/NF-κB信号通路

Key words:  Xuebijing Injection  sepsis  acute kidney injury  TLR4/MyD88/NF-κB signaling pathway

摘要:

[目的] 探讨血必净注射液（XBJ）通过Toll样受体4（TLR4）/髓样分化因子88（MyD88）/核转录因子-κB（NF-κB）信号通路保护脓毒症诱导的急性肾损伤大鼠作用机制。[方法] 将18只SD大鼠随机分为假手术组、模型组和血必净组，每组6只。计算肾脏指数，通过苏木精-伊红（HE）染色确定肾组织病理学改变情况。通过酶联免疫吸附实验（ELISA）试剂盒检测肌酐（Cr）、血尿素氮（BUN）、中性粒细胞明胶酶相关脂质运载蛋白（NGAL）、肾损伤分子1（Kim-1）、肿瘤坏死因子α（TNF-α）和白细胞介素-6（IL-6）的表达水平。采用蛋白印迹方法对TLR4、MyD88、p-NF-κB、B淋巴细胞瘤-2蛋白（Bcl-2）、Bcl-2相关X蛋白（Bax）和活化的半胱氨酸天冬氨酸蛋白酶-3（Cleaved Caspase 3）的表达水平进行检测。[结果] 与模型组相比，血必净组可以有效降低Cr、BUN、NGAL、Kim-1、TNF-α和IL-6的表达水平，差异具有统计学意义（P＜0.05），同时减轻了脓毒症诱导急性肾损伤大鼠的病理损伤程度和肾脏指数。同时，与模型组相比，给予XBJ治疗后可以明显降低Bax和Cleaved Caspase 3的表达水平，增加Bcl-2蛋白的表达水平，差异具有统计学意义（P＜0.05）。此外，与模型组相比，给予XBJ治疗后可以明显降低TLR4、MyD88和p-NF-кB蛋白的表达水平，差异具有统计学意义（P＜0.05）。[结论] XBJ连续治疗可以减轻脓毒症诱导急性肾损伤程度并可以改善肾功能，其作用机制可能与抑制TLR4/MyD88/NF-κB信号通路减轻炎症反应和细胞凋亡有关。

Abstract:

[Objective] To investigate the protective mechanism of Xuebijing Injection（XBJ） against sepsis-induced acute kidney injury through TLR4/MyD88/NF-κB signaling pathway in rats. [Methods] Eighteen SD rats were randomly divided into Sham operation group，model group and Xuebijing group，with 6 rats in each group. The renal index was calculated，and the pathological changes of renal tissue were determined by hematoxylin-eosin（HE） staining. The expression levels of creatinine（Cr），blood urea nitrogen（BUN），neutrophil gelatinase associated lipocalin（NGAL），kidney injury molecule-1（Kim-1），tumor necrosis factor-α（TNF-α） and interleukin-6（IL-6） were detected by ELISA kits. The expression levels of TLR4，MyD88，p-NF-κB，Bcl-2，Bax and Cleaved Caspase 3 were detected by Western blot. [Results] Compared with the model group，the Xuebijing group could effectively reduce the expression levels of Cr，BUN，NGAL，Kim-1，TNF-α and IL-6，and the difference was statistically significant（P＜0.05）. At the same time，the Xuebijing group reduced the degree of pathological injury and renal index in the rats with sepsis-induced acute kidney injury. At the same time，compared with the model group，XBJ treatment could significantly reduce the expression levels of Bax and Cleaved Caspase 3，and increase the expression level of Bcl-2 protein，the difference was statistically significant（P＜0.05）. In addition，XBJ treatment significantly reduced the protein expression levels of TLR4，MyD88 and p-NF-κB compared with model group（P＜0.05）. [Conclusion] XBJ continuous treatment can reduce the degree of sepsis-induced acute kidney injury and improve renal function，and its mechanism may be related to the inhibition of TLR4/MyD88/NF-κB signaling pathway to reduce inflammatory response and apoptosis.