基于SDF-1/CXCR4信号通路探讨新安鼻渊方流浸膏对急性鼻窦炎大鼠嗅上皮神经元细胞凋亡的影响

The effect of xinan nasoyuan fangliu extract on the apoptosis of olfactory neurons in rats with acute sinusitis based on SDF-1/CXCR4 signaling pathway

目的基于SDF-1/CXCR4信号通路探讨新安鼻渊方流浸膏对急性鼻窦炎大鼠嗅上皮神经元细胞凋亡的影响。方法 40只SD大鼠分为对照组,模型组,治疗组和抑制剂组,每组10只。除对照组外,其余各组大鼠采用金黄色葡萄球菌悬液注射法构建急性鼻窦炎模型,对照组予以无菌生理盐水代替。造模后第7天开始采用灌胃方式连续给药1周,每天1次,治疗组给药新安鼻渊方流浸膏按照大鼠体重计算(7 g/kg),抑制剂组给予1mg/kg CXCR4特异性抑制剂AMD3100,对照组以及模型组灌胃等量的生理盐水。观测大鼠鼻窦炎病理特征并进行评分以鉴定模型的建立,通过酶联免疫法检测各组大鼠嗅上皮组织的炎性因子TNF-α以及IL-1β的表达情况,通过TUNEL法检测各组大鼠嗅上皮组织中细胞凋亡数量,免疫组化检测SDF-1、CXCR4的表达,采用Western blot检测各组大鼠嗅上皮神经元细胞中促凋亡蛋白cleaved caspase 3、Bax以及抗凋亡蛋白Bcl-2的表达。结果模型组大鼠鼻窦炎病理评分显著高于对照组(P<0.05),治疗组相比与模型组显著降低(P<0.05)。与对照组相比,模型组的炎性因子表达明显升高(P<0.05),治疗组显著低于模型组(P<0.05)。TUNEL染色结果显示:与对照组相比,模型组大鼠嗅上皮神经元细胞凋亡显著增加(P<0.05),治疗组以及抑制剂组大鼠嗅上皮神经元细胞的凋亡显著低于模型组(P<0.05)。与对照组相比,模型组抗凋亡蛋白Bcl-2、SDF-1以及CXCR4的表达明显降低(P<0.05),促凋亡蛋白cleaved caspase 3、Bax显著增加(P<0.05),治疗组相比于模型组抗凋亡蛋白Bcl-2、SDF-1以及CXCR4的表达明显增加(P<0.05),促凋亡蛋白cleaved caspase 3、Bax显著降低(P<0.05)。结论新安鼻渊方流浸膏可改善急性鼻窦炎大鼠嗅上神经元细胞的凋亡,该作用机制可能与抑制SDF-1/CXCR4信号通路密切相关。

Objective To investigate the effect of xinan nasoyuan fangliu extract on the apoptosis of olfactory neurons in rats with acute sinusitis and the role of the SDF-1/CXCR4 signaling pathway.Methods Forty Sprague Dawley rats were divided into a control group,model group,treatment group and inhibitor group,with 10 rats in each group.The rats in all groups except the control group were treated with staphylococcus aureus suspension to establish the acute sinusitis model,and the control group received sterile saline.On the seventh day after the model was established,treatment administration was conducted by gavage every day for 1 week continuously.The treatment group was given xin’an nasoyuan fangliu extract according to body weight(7 g/kg),the inhibitor group received 1 mg/kg CXCR4 specific inhibitor AMD3100,and the control group and the model group were given the same amount of normal saline.Pathological features of sinusitis were observed and scored to confirm the establishment of the model.Enzyme-linked immunoassay was used to detect the expression of inflammatory cytokines TNF-αand IL-1βin the olfactory epithelium tissue.TUNEL staining,to detect the number of apoptotic cells,and immunohistochemistry of SDF-1 and CXCR4 were performed in olfactory epithelial tissue of rats in each group.Western blot was used to assess the expression of pro-apoptotic proteins,such as cleaved caspase 3 and Bax,and anti-apoptotic protein Bcl-2 in neurons of the olfactory epithelium.Results The pathological score of sinusitis in the model group was significantly higher than that in the control group(P<0.05),and that in the treatment group was significantly lower than that in the model group(P<0.05).The expression of inflammatory factors in the model group was significantly increased compared with that in the control group(P<0.05),and that in the treatment group was significantly lower than that in the model group(P<0.05).TUNEL staining showed that olfactory epithelial neuron apoptosis increased in the model group compared with the control group(P<0.05),and was significantly lower in the treatment group and inhibitor group than in the model group(P<0.05).In the model group,Bcl-2,SDF-1 and CXCR4 expressions were significantly decreased(P<0.05),whereas cleaved caspase 3 and Bax expressions were significantly increased(P<0.05)compared with the control group.Bcl-2,SDF-1 and CXCR4 expressions in the treatment group were significantly increased compared with those in the model group(P<0.05),whereas cleaved caspase 3 and Bax expressions were significantly decreased in the treatment group(P<0.05).Conclusions Xinan nasoyuan fangliu extract improved apoptosis of rat superior olfactory neurons in acute sinusitis,and the effect may be mediated by inhibition of the SDF-1/CXCR4 signaling pathway.