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目的:研究痰热清注射液对Lewis肺癌化疗模型小鼠免疫抑制作用的影响,以探讨其对荷肺癌小鼠的免疫调节作用。方法:构建C57 BL/6小鼠Lewis肺癌模型,40只模型小鼠随机分为模型对照组、痰热清组、化疗组、痰热清联合化疗组及正常对照组,每组8只。治疗后摘小鼠眼球取血,取脾脏、外周血和胸腺,并分离腹腔巨噬细胞(Mφ),检测各组小鼠胸腺指数及外周血淋巴细胞数量,MTT法检测各组小鼠脾T、B淋巴细胞增殖水平,流式细胞术检测胸腺细胞的凋率和外周血内CD3+、CD3-NK1.1+细胞及CD3+NK1.1+细胞比例,RT-PCR检测胸腺组织Bcl-2和Fas mRNA表达水平,乳酸脱氢酶(LDH)释放法和ELISA法检测Mφ的杀伤活性及及其分泌IL-12能力,免疫透射比浊法检测小鼠外周血清中Ig G及补体C3的含量。结果:痰热清联合化疗组小鼠的胸腺指数、外周血淋巴细胞计数及T、B细胞增殖活性均明显高于单独化疗组(P<0.01),同时胸腺细胞的凋亡水平明显低于单独化疗组(P<0.01)。痰热清联合化疗组小鼠外周血CD3+T细胞、CD3-NK1.1+细胞及CD3+NK1.1+细胞比例均明显高于化疗组(P<0.01)。与化疗组相比,痰热清联合化疗组小鼠胸腺组织Bcl-2 mRNA水平明显增高,而Fas mRNA水平明显减低(均P<0.01)。痰热清组小鼠Mφ杀伤活性及分泌IL-12水平均明显高于化疗组(均P<0.01),同时外周血Ig G及补体C3含量明显高于化疗组(均P<0.01)。结论:痰热清注射液对肺癌化疗小鼠的免疫系统有明显保护作用,其对抗肿瘤免疫力有明显的促进作用。
Objective: To study the effect of Tanreqing injection on the immunosuppressive effect of Lewis lung cancer model mice to investigate its immunomodulatory effect on mice bearing lung cancer. Methods: Lewis lung carcinoma model was established in C57 BL / 6 mice. Forty mice were randomly divided into model control group, Tanreqing group, chemotherapy group, Tanreqing combined chemotherapy group and normal control group, with 8 mice in each group. After treatment, the eyeballs of mice were taken for blood, spleens, peripheral blood and thymus were taken, and the peritoneal macrophages (Mφ) were isolated. The thymus index and the number of peripheral blood lymphocytes in each group were detected. , The proliferation of B lymphocytes, the apoptosis of thymocytes and the proportion of CD3 +, CD3-NK1.1 + cells and CD3 + NK1.1 + cells in peripheral blood were measured by flow cytometry. The levels of Bcl-2 and Fas mRNA expression, lactate dehydrogenase (LDH) release assay and ELISA assay were used to detect the Mφ cytotoxicity and IL-12 secretion ability. The levels of Ig G and complement C3 in peripheral blood were detected by immunoturbidimetry. Results: The thymus index, peripheral blood lymphocyte count and T, B cell proliferative activity in Tanreqing combined with chemotherapy group were significantly higher than those in chemotherapy alone group (P <0.01), while the thymocyte apoptosis level in Tanreqing combined chemotherapy group was significantly lower than that in the chemotherapy alone group Chemotherapy group (P <0.01). The proportion of CD3 + T cells, CD3-NK1.1 + cells and CD3 + NK1.1 + cells in the Tanreqing combined with chemotherapy group were significantly higher than those in the chemotherapy group (P <0.01). Compared with chemotherapy group, the levels of Bcl-2 mRNA and the level of Fas mRNA in thymus tissue of Tanreqing combined with chemotherapy group were significantly decreased (all P <0.01). The Mφ cytotoxic activity and IL-12 secretion of Tanreqing group were significantly higher than that of chemotherapy group (all P <0.01), while the contents of Ig G and complement C3 in peripheral blood were significantly higher than those in chemotherapy group (all P <0.01). Conclusion: Tanreqing injection has obvious protective effect on the immune system of mice with chemotherapy of lung cancer, which has a significant promotion on anti-tumor immunity.