苦酒汤配方工艺优化及其药效机制

Formulation Process Optimization and Pharmacodynamic Mechanism of Bitter Wine Soup

关键词:  苦酒汤  工艺优化  16HBE细胞  慢性咽炎

Key words:  Bitter Wine Soup  process optimization  16HBE cells  chronic pharyngitis

摘要:

目的 优化苦酒汤加工工艺与配方，并探究其药效及作用机制。方法 采用单因素结合正交试验对苦酒汤配方进行优化，以抑菌圈平均直径作为苦酒汤配方优化评价指标。制备空白组，模型组，阳性药组，苦酒汤低、中、高剂量组及苦酒汤古方组大鼠含药血清，体外培养人支气管上皮细胞(16HBE)，以20 μg·mL^-1的脂多糖及10%含药血清进行干预，CCK-8法检测各组含药血清对脂多糖诱导的16HBE细胞增殖活力的影响，ELISA检测细胞上清液中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)和白细胞介素1β(IL-1β)的含量，PCR检测P65、IκBα mRNA的表达。结果 苦酒汤最优配方为米醋60 mL，鸡蛋清30 g，清半夏10 g，凤凰衣1 g。与空白组比较，模型组16HBE增殖活力下降(P<0.01)，TNF-α、IL-6和IL-1β含量升高(P<0.01)，P65 mRNA表达上调，IκBα mRNA表达下调(P<0.01)。与模型组比较，苦酒汤中、高剂量组及古方组细胞增殖活力上升(P<0.01)，TNF-α、IL-6、IL-1β含量下降(P<0.01)，P65 mRNA表达下调，IκBα mRNA表达上调(P<0.01)。与苦酒汤古方组比较，苦酒汤高剂量组TNF-α、IL-1β含量下降(P<0.01)，P65 mRNA表达下调(P<0.01)。结论 经工艺优化后的苦酒汤制备方便，用量明确，其含药血清能够减轻脂多糖诱导的16HBE细胞炎症反应，药效优于原方，其作用机制可能与抑制NF-κB信号通路激活有关。

Abstract:

OBJECTIVE To optimize the processing technology and formula of Bitter Wine Soup, and to explore its efficacy and mechanism. METHODS The formula of Bitter Wine Soup was optimized by a single factor combined with an orthogonal test, and the average diameter of the inhibition zone was used as the evaluation index for formula optimization of Bitter Wine Soup. Prepare control group, model group, positive drug group, Bitter Wine Soup low, medium and high dose groups and Bitter Wine Soup ancient prescription group rat drug-containing serum, cultured human bronchial epithelial cells(16HBE) in vitro, intervened with 20 μg·mL^-1 lipopolysaccharide and 10% drug-containing serum of different groups, the effect of drug-containing serum on lipopolysaccharide-induced 16HBE cell proliferation was detected by CCK-8 method, and the contents of tumor necrosis factor α (TNF-α), interleukin 6(IL-6), interferon-1β(IL-1β) of leukocytes in cell supernatant were detected by ELISA, the expression of P65 and IκBα mRNA was detected by PCR. RESULTS The optimal formula for Bitter Wine Soup was as follows: 60 mL rice vinegar, 30 g egg white, 10 g Pinelliae Rhizoma Praeparatum Cum Alumine, and 1 g Membrana Follicularisovi. Compared with the control group, the proliferation activity of 16HBE in the model group was decreased(P<0.01), the contents of TNF-α, IL-1β and IL-6 were increased(P<0.01), the expression of P65 mRNA was up-regulated, and the expression of IκBα mRNA was down-regulated(P<0.01). Compared with the model group, the cell proliferation activity in the Bitter Wine Soup medium and high dose groups and the ancient prescription group increased(P<0.01), the contents of TNF-α, IL-1β and IL-6 decreased(P<0.01), the expression of P65 mRNA was down-regulated, and the mRNA of IκBα expression was up-regulated(P<0.01). Compared with the Bitter Wine Soup ancient prescription group, the contents of TNF-α and IL-1β in the Bitter Wine Soup high-dose group decreased(P<0.01), and the expression of P65 mRNA was down-regulated(P<0.01).CONCLUSION The optimized formulation and processing technology of Bitter Wine Soup is convenient for preparation, and the dossage is clear. Its drug-containing serum can alleviate lipopolysaccharide-induced 16HBE inflammatory response, and its efficacy is better than that of the original formula. Its mechanism may be related to the inhibition of NF-κB signaling pathway.