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目的研究通窍止鼽汤对阻断变应性鼻炎(allergic rhinitis,AR)SD大鼠PKC-NF-κB信号传导系统的作用,探讨其治疗AR的作用机制。方法在2015年9月—2016年2月选取4月龄健康SD大鼠80只,雌雄各半,体重180 g~220 g,随机分为正常对照组、正常PMA组、正常PMA+PDTC组、正常PMA+中药组、AR对照组、AR PMA组、AR PMA+PDTC组、AR PMA+中药组,每组10只,共8组。采用卵清蛋白和热灭活百日咳杆菌为致敏原制备SD大鼠行实验性AR模型。造模成功后,正常PMA+中药组和AR PMA+中药组灌服中药液17.5 g/kg(生药浓度2 g/ml),余6组均灌服生理盐水2 ml/只,1次/d,共14 d。采用尼龙毛法分离T淋巴细胞,并进行体外培养。T淋巴细胞涂片免疫组化染色测定NF-κB的表达,ELISA法测定大鼠T淋巴细胞培养液上清液白细胞介素(interleukin,IL)-4和IL-5含量。多组间均数比较采用单因素方差分析,两变量的相关程度用直线相关分析法分析,P<0.05为差异有统计学意义。结果正常各组T淋巴细胞核转录因子-κB(nuclear factor-kappa B,NF-κB)细胞核染色阳性百分比和培养液上清液IL-4和IL-5含量比较差异均无统计学意义(均P>0.05);正常对照组与AR对照组比较,差异有统计学意义(P<0.05)。AR PMA组T淋巴细胞NF-κB细胞核染色阳性百分比和培养液上清液IL-4和IL-5含量明显升高,与AR PMA+PDTC组、AR PMA+中药组比较差异均有统计学意义(均P<0.05),AR PMA+PDTC、AR PMA+中药组均能明显降低淋巴细胞NF-κB细胞核染色阳性百分比和培养液上清液IL-4和IL-5含量,对比差异均有统计学意义(均P<0.05),以AR PMA+中药组效果最佳。各组T淋巴细胞NF-κB细胞核染色阳性百分比与培养液上清液IL-4、IL-5含量成显著的正相关性(r1=0.998、r2=0.993,均P<0.05)。结论通窍止鼽汤通过阻断AR大鼠PKC-NF-κB信号传导系统,减少Th2细胞因子的产生,从而抑制Th2细胞的免疫应答优势,纠正Th1/Th2的失衡状态,达到治疗AR的目的。
Objective To investigate the effect of Tongqiao Zhilu Decoction on blocking PKC-NF-κB signaling system in allergic rhinitis (SD) rats and to explore its mechanism of action in treating AR. Methods From September 2015 to February 2016, 80 healthy 4-month-old SD rats were randomly divided into normal control group, normal PMA group, normal PMA + PDTC group, Normal PMA + Chinese medicine group, AR control group, AR PMA group, AR PMA + PDTC group, AR PMA + Chinese medicine group, with 10 in each group. The experimental AR model of SD rats was prepared by ovalbumin and heat-inactivated Bordetella pertussis. After successful modeling, the normal PMA + Chinese medicine group and the AR PMA + Chinese medicine group were given 17.5 g / kg Chinese medicine solution (crude drug concentration 2 g / ml), and the other 6 groups were given normal saline 2 ml once a day, A total of 14 d. T lymphocytes were isolated by nylon wool method and cultured in vitro. The expression of NF-κB was detected by T lymphocyte smear immunohistochemical staining. The levels of interleukin (IL) -4 and IL-5 in supernatant of T lymphocyte were measured by ELISA. One-way ANOVA was used to compare the mean of multiple groups, and the correlation between the two variables was analyzed by linear correlation analysis. P <0.05 was considered statistically significant. Results The positive percentage of nuclear staining of nuclear factor-kappa B (NF-κB) and the content of IL-4 and IL-5 in supernatant of normal group were not significantly different > 0.05). There was significant difference between normal control group and AR control group (P <0.05). The percentage of nuclear staining of NF-κB and the level of IL-4 and IL-5 in supernatant of AR PMA group were significantly higher than those of AR PMA + PDTC group and AR PMA + All P <0.05). Both AR PMA + PDTC and AR PMA + Chinese herbs group could significantly reduce the positive percentage of nuclear staining of NF-κB and the content of IL-4 and IL-5 in supernatant of culture supernatant, the differences were statistically significant (All P <0.05), and AR PMA + Chinese medicine group had the best effect. The positive percentage of nuclear staining of NF-κB in each group was positively correlated with the contents of IL-4 and IL-5 in supernatant of culture supernatant (r1 = 0.998, r2 = 0.993, all P <0.05). Conclusion Tongqiao Zhixue decoction can inhibit the immune response of Th2 cells by blocking the PKC-NF-κB signal transduction system of AR rats and reduce the production of Th2 cytokines, and correct the imbalance of Th1 / Th2 to achieve the goal of AR.