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【目的】评价冰香散挥发油抗流感病毒黏膜免疫的效果。【方法】将30只BABL/c小鼠分为正常对照组、安慰剂组、以及冰香散挥发油组,每组10只。提前7 d预防给药,正常对照组正常饲养,安慰剂组(体积分数为1%吐温-80)灌胃给药(1 m L/d),冰香散挥发油组滴鼻给药(0.05 m L/d),以5倍半数致死量(5LD50)流感病毒液A/FM/1/47(H1N1)滴鼻感染小鼠,建立病毒性肺炎小鼠模型。之后连续观察5 d,眼球摘除取血,收集鼻黏膜、支气管肺泡灌洗液,采用酶联免疫吸附试验(ELISA)方法检测Ig A、Ig G、分泌型Ig A(s-Ig A)含量;分离鼻相关淋巴组织,采用免疫组织化学法测定CD4、CD8表达;收集脾淋巴细胞,以刀豆蛋白(Con A)和脂多糖(LPS)刺激,采用细胞增殖与活性试剂盒(CCK-8)检测脾淋巴细胞增殖情况;采用ELISA检测脾淋巴细胞分泌细胞因子干扰素-γ(IFN-γ)、白细胞介素-4(IL-4)水平。【结果】冰香散挥发油组能提高灌洗液s-Ig A和血清Ig A水平,提高血清Ig G含量,刺激CD4+、CD8+T细胞分化与成熟,刺激脾淋巴细胞增殖,刺激细胞因子IFN-γ、IL-4分泌,与安慰剂组比较,差异均有统计学意义(P<0.05或P<0.01)。【结论】冰香散挥发油滴鼻免疫能够有效地诱导黏膜免疫反应,引起体液免疫和细胞免疫,发挥抗病毒作用。
【Objective】 To evaluate the anti-influenza virus mucosal immunity of Bingxiao San volatile oil. 【Methods】 Thirty BABL / c mice were divided into normal control group, placebo group, and Bingxian San volatile oil group, with 10 mice in each group. The rats in the normal control group were fed with normal saline for 7 days and the normal control group was given normal saline (1 m L / d) in the placebo group (1% Tween-80) mice were infected intranasally with 5LD50 of influenza virus A / FM / 1/47 (H1N1) by intraperitoneal injection of 5x50 of Ml / d. The mouse model of viral pneumonia was established. After continuous observation for 5 days, the eyes were removed for blood collection. Nasal mucosa and bronchoalveolar lavage fluid were collected and the contents of Ig A, Ig G and s-Ig A were detected by enzyme linked immunosorbent assay (ELISA) Nasal lymphoid tissue was isolated and the expression of CD4 and CD8 was detected by immunohistochemistry. Splenic lymphocytes were collected and stimulated with Con A and LPS. Cell proliferation and activity kit (CCK-8) The proliferation of splenic lymphocytes was detected. The levels of IFN-γ and IL-4 secreted by splenic lymphocytes were detected by ELISA. [Result] Bingxiao San volatile oil group can improve the level of s-Ig A and IgA in serum, increase the content of serum Ig G, stimulate the differentiation and maturation of CD4 + and CD8 + T cells, stimulate the proliferation of splenic lymphocytes and stimulate the secretion of cytokines IFN -γ, IL-4 secretion compared with placebo group, the difference was statistically significant (P <0.05 or P <0.01). 【Conclusion】 Bingxiao San volatile oil intranasal immunization can effectively induce mucosal immune response, causing humoral and cellular immunity, play an antiviral effect.