茯苓四逆汤调节TLR-4/NF-κB 通道对脓毒症大鼠心肌抑制的保护机制研究

Protective Mechanism of Fuling Sini Decoction on Myocardial Depression in Septic Rats Via TLR-4/NF-κB Signaling Pathway

目的:探讨茯苓四逆汤调节TLR-4/NF-κB通道对脓毒症大鼠心肌抑制的保护作用机制。方法:SPF级SD雄性大鼠120只,随机分成6组:正常组、假手术组、模型组,中药低、中、高剂量组。通过盲肠结扎穿孔法(CLP法)制作脓毒症模型。造模后6h开始给药,持续给药3d。正常组、假手术组、模型组予等量生理盐水,中药各剂量组予相应剂量茯苓四逆汤,均为灌胃给药。观察大鼠生存状态,于术前和术后24h、48h、72h各选5只大鼠处死,采用Western blotting法及QPCR检测心肌组织NF-κB、H-FABP的蛋白和TLR-4的基因表达,同时以ELISA法检测心肌细胞TNF-ɑ、IL-6含量。结果:与正常组比较,模型组及中药各剂量组TNF-ɑ、IL-6水平及NF-κB、H-FABP蛋白表达明显升高,TLR-4 基因表达亦增高;与模型组同时间点比较,中药各剂量组TNF-ɑ、IL-6水平明显下降,NF-κB 和H-FABP蛋白水平比较也有统计学差异;中药各剂量组同时间比较发现,剂量越高,其TNF-ɑ、IL-6水平下降越明显,下调NF-κB 和H-FABP蛋白表达水平越明显,TLR-4 mRNA 转录水平降低的越明显,差异均有统计学意义。结论:茯苓四逆汤能减轻脓毒症大鼠心肌抑制,通过抑制炎症反应,负反馈调节TLR-4/NF-κB信号通路,从而缓解心肌抑制。

Objective:To investigate the protective effect of Fuling Sini Decoction on myocardial depression in septic rats by regulating TLR-4/NF-κB channel.Methods:120 SPF male SD rats were randomly divided into 6 groups:normal group,sham operation group,model group,and low,medium,and high-dose Chinese medicine groups.The sepsis model was made by the cecal ligation and perforation method (CLP method).The administration was started 6 hours after the model was established,and the administration was continued for 3 days.The normal group,sham operation group,and model group were given the same amount of normal saline,and each dose group of Chinese medicine was given the corresponding dose of Fulingsini Decoction,all of which were given by intragastric administration.The survival status of the rats was observed.Five rats were selected and sacrificed at 24h,48h,72h before and after the operation.Western blotting and QPCR were used to detect the protein of NF-κB and H-FABP and the gene expression of TLR-4 in myocardial tissue.At the same time,ELISA was used to detect the content of TNF-ɑ and IL-6 in cardiomyocytes.Results:Compared with the normal group,the levels of TNF-ɑ,IL-6 and the protein expression of NF-κB and H-FABP in the model group and each dose group of traditional Chinese medicine increased significantly,and the expression of TLR-4 gene also increased;at the same time point as the model group In comparison,the levels of TNF-ɑ and IL-6 in each dose group of traditional Chinese medicine decreased significantly,and the protein levels of NF-κB and H-FABP were also statistically different.Comparing the different dose groups of traditional Chinese medicine at the same time,it was found that the higher the dose,the higher the level of TNF-ɑ,The more obvious the decrease of IL-6 level,the more obvious the down-regulation of NF-κB and H-FABP protein expression level,the more obvious the decrease of TLR-4 mRNA transcription level,and the difference is statistically significant.Conclusion:Fuling Sini Decoction can alleviate myocardial depression in rats with sepsis.By inhibiting the inflammatory response,negative feedback regulates the TL R-4/NF-κB signaling pathway,thereby alleviating myocardial depression.