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目的:观察小檗碱和α2肾上腺素能受体拮抗剂育亨宾对内毒素血症小鼠脾脏Toll样受体4(TLR4)信号通路84种基因表达的影响,并初步探讨其作用机制。方法:雄性BALB/c小鼠随机分为对照组、脂多糖(LPS)组、小檗碱+LPS组、小檗碱+育亨宾+LPS组、育亨宾+LPS组、小檗碱组、小檗碱+育亨宾组和育亨宾组。分别用蒸馏水、小檗碱(50 mg/kg)、小檗碱+育亨宾(50 mg/kg+2 mg/kg)和育亨宾(2 mg/kg)灌胃,每天1次,连续3 d,第3 d灌胃1 h后,腹腔注射LPS(20 mg/kg)或生理盐水。腹腔注射1 h后,用RT2ProfilerTMPCR Array分析技术检测小鼠脾脏TLR4信号通路84种基因mRNA的表达;用Western blotting分析小鼠脾脏TLR4信号通路的抑制分子细胞因子信号抑制物(SOCS)1、SOCS3和白细胞介素-1受体相关激酶(IRAK)-M蛋白的表达。结果:LPS可上调小鼠脾脏TLR4信号转导通路中相关炎症因子的mRNA表达,包括CXCL10、TNF-α、IL-1α、IL-1β、IL-6、IFN-γ和IFN-β。小檗碱能显著下调下调髓样分化因子(MyD88)依赖信号通路下游TNF-α、IL-1α、IL-1β和IL-6 mRNA的表达,也能MyD88非依赖信号通路下游基因IFN-β和CXCL10 mRNA的表达(P<0.05)。育亨宾能显著下调内毒素血症小鼠脾脏IL-1α、IL-1β和IFN-βmRNA的表达(P<0.05),但对TNF-α、IL-6和CXCL10 mRNA表达的下调作用与LPS组相比没有显著差异(P>0.05)。小檗碱与育亨宾合剂能显著下调内毒素血症小鼠脾脏IFN-β和CXCL10 mRNA的表达,但不能显著下调内毒素血症小鼠脾脏IL-1α、IL-1β、TNF-α和IL-6 mRNA的表达。LPS攻击后1 h,小檗碱和(或)育亨宾均不能增强内毒素血症小鼠脾脏SOCS1、SOCS3和IRAK-M蛋白的表达。结论:小檗碱和育亨宾均能抑制LPS诱导的MyD88依赖和非依赖信号通路下游部分基因的表达,这种抑制作用的机制与SOCS1、SOCS3和IRAK-M蛋白无关。
OBJECTIVE: To observe the effect of yohimbine, a berberine and α2 adrenergic receptor antagonist, on the expression of 84 genes in Toll-like receptor 4 (TLR4) signaling pathway of endotoxemia mice and to explore its possible mechanism. Methods: Male BALB / c mice were randomly divided into control group, lipopolysaccharide (LPS) group, berberine + LPS group, berberine + yohimbine + LPS group, yohimbine + LPS group and berberine group , Berberine + yohimbine group and yohimbine group. The rats were orally administered with distilled water, berberine (50 mg / kg), berberine plus yohimbine (50 mg / kg + 2 mg / kg) and yohimbine (2 mg / On the 3rd day, LPS (20 mg / kg) or normal saline was injected intraperitoneally 1 h after the third day. One hour after intraperitoneal injection, the mRNA expression of 84 genes in TLR4 signaling pathway in spleen of mice was detected by RT2 Profiler TMPCR Array analysis. The inhibitory molecular cytokine signaling inhibitor (SOCS) 1, SOCS3 and Interleukin-1 receptor associated kinase (IRAK) -M protein expression. Results: LPS up-regulated the mRNA expression of related inflammatory factors, including CXCL10, TNF-α, IL-1α, IL-1β, IL-6, IFN-γ and IFN-β in TLR4 signal transduction pathway in mouse spleen. Berberine down-regulated the expression of TNF-α, IL-1α, IL-1β and IL-6 mRNA downstream of MyD88-dependent signaling pathway, and down-regulated MyD88-dependent signaling pathway IFN- CXCL10 mRNA expression (P <0.05). Yohimbin could significantly down-regulate the expression of IL-1α, IL-1β and IFN-βmRNA in the spleens of endotoxemia mice (P <0.05), but down-regulated the expressions of TNF-α, IL-6 and CXCL10 mRNA in LPS There was no significant difference between groups (P> 0.05). Berberine and Yohimbin could significantly down-regulate the expression of IFN-β and CXCL10 mRNA in the spleen of endotoxemic mice, but could not significantly down-regulate the levels of IL-1α, IL-1β, TNF-α and IL-6 mRNA expression. At 1 h after LPS challenge, neither berberine nor yohimbine enhanced the expression of SOCS1, SOCS3 and IRAK-M in spleen of endotoxemic mice. CONCLUSION: Both berberine and yohimbine can inhibit the LPS-induced expression of some genes downstream of MyD88-dependent and independent signaling pathway. The mechanism of this inhibitory effect is not related to SOCS1, SOCS3 and IRAK-M proteins.