益肺消瘤颗粒延缓免疫衰老防治肺癌的实验研究

Experimental Study of Yifeixiaoliu Granulesto Delays Immunosenescence and Prevents Senile Lung Cancer

目的:探讨益肺消瘤颗粒对免疫衰老小鼠肺癌移植瘤的防治作用。方法:C57BL/6J雄性小鼠随机分为空白组,生理盐水组和益肺消瘤组,益肺消瘤颗粒组、生理盐水组小鼠予以D-半乳糖500 mg/(kg∙d)皮下注射建立免疫衰老模型,连续42天,同时益肺消瘤组小鼠每天灌服益肺消瘤颗粒52 g/kg/d (终体积0.2 mL),空白组及生理盐水组小鼠每天灌服等体积的生理盐水。检测小鼠血清超氧化物歧化酶活力和丙二醛的含量,摘取小鼠胸腺,计算器官指数;流式细胞术检测小鼠胸腺T细胞免疫衰老相关膜分子表达。再次C57BL/6J雄性建立免疫衰老小鼠模型,左侧腋下前部接种肺癌细胞LL2-Luc-M38建立肺癌皮下移植瘤模型,并随机分为生理盐水组、益肺消瘤颗粒预防组和益肺消瘤颗粒防治组进行干预,每组取10只小鼠予以移植瘤造模,14 d后处死,检测瘤体质量;其余小鼠观察成瘤时间、计算生存期。结果:1) 与空白组比较,生理盐水组小鼠胸腺指数下降(P < 0.01),血清SOD活力明显下降(P < 0.01),MDA含量明显上升(P < 0.01),CD3+CD25+、CD3+CD28+、CD3+CD45RA+的表达下降(P < 0.001或P < 0.01),CD3+CD196+、CD4+CD25+的表达上升(P < 0.01或0.05);益肺消瘤颗粒干预后,与生理盐水组比较,益肺消瘤颗粒组小鼠胸腺指数增加(P < 0.05),血清SOD活力明显增加(P < 0.01),MDA含量明显下降(P < 0.01),CD3+CD25+、CD3+CD28+、CD3+CD45RA+的表达上升(P < 0.01或P < 0.05),CD3+CD196+、CD4+CD25+的表达下降(P < 0.05);2) 免疫衰老小鼠皮下接种肺癌细胞后,益肺消瘤颗粒预防组小鼠的成瘤时间长于生理盐水组(P < 0.05),益肺消瘤颗粒防治组的成瘤时间、生存期延长更为明显(P < 0.01或P < 0.05),益肺消瘤颗粒预防组移植瘤体质量明显小于生理盐水组(P < 0.05)和防治组(P < 0.01),益肺消瘤颗粒预防组的成瘤时间小于防治组(P < 0.05)、瘤重大于防治组(P < 0.05)。结论:D-半乳糖可诱导小鼠免疫衰老发生;益肺消瘤颗粒可延缓小鼠免疫衰老,从而防治肺癌的发生发展及延长小鼠生存期。

Objective: To study the preventive and therapeutic effect of Yifeixiaoliu Granules on lung carcinoma in immunosenescent miceinduced. Methods: C57BL/6J male mice were randomly divided into blank group, normal saline solutiongroup and Yifeixiaoliu Granules group, C57BL/6J male mice in Yifeixiaoliu granules group and normal saline solution group were subcutaneously injected with 500 mg/(kg∙d) D-galactose to establish an immuneosenescent mice for 42 consecutive days. At the same time, C57BL/6J male mice in Yifeixiaoliu granules group were given 52 g/kg/d (final volume 0.2 mL) of YifeiXiaooma granule every day, while C57BL/6J male mice in blank group and normal saline group were given the same volume of normal saline every day. The activity of SOD and the content of malondialdehyde MDA in serum of mice were detected, mouse thymus was extracted and organ index was calculated. Flow cytometry was used to detect the expression of immunosenescence related membrane molecules in mouse thymus T cells. Again, C57BL/6J male mice model of immunosenescence was established, and LL2-Luc-M38 was inoculated in the left anterior armpit to establish the subcutaneous lung carcinoma transplant tumor model, and they were randomly divided into normal saline solution group, Yifeixiaoliu granules prevention group and Yifexiaoma granule prevention treatment group, 10 mice from each group were taken for tumor transplantation model 14 days later, they were killed. Tumor mass was detected. Tumor formation time and survival time of other mice were observed. Results: 1) Compared with blank group, the thymus index of mice in normal saline solutiongroup was decreased (P < 0.01), the activity of SOD in serum was decreased (P < 0.01), and the content of MDA was increased (P < 0.01). The expressions of CD3+CD25+, CD3+CD28+ and CD3+CD45RA+ were decreased (P < 0.001 or P < 0.01), while the expressions of CD3+CD196+ and CD4+CD25+ were increased (P < 0.01 or 0.05). After an intervention of Yifeixiaoliu Granules, compared with normal saline group, the thymus index of mice in Yifei Xiaoliu granules group was increased (P < 0.05), the activity of SOD in serum was significantly increased (P < 0.01), and the content of MDA was significantly decreased (P < 0.01). The expressions of CD3+CD25+, CD3+CD28+ and CD3+CD45RA+ were increased (P < 0.01 or P < 0.05), while the expressions of CD3+CD196+ and CD4+CD25+ were decreased (P < 0.05). 2) After subcutaneous inoculation of lung carcinoma cells in immunosenescent mice, the tumor formation time of mice in Yifeixiaoliu granules prevention group was longer than that in normal saline solution group (P < 0.05). The tumor formation time and survival time of Yifeixiaoliu granules prevention treatment group were more significantly extended (P < 0.01 or P < 0.05). The tumor transplantation weight of Yifeixiaoliu granules prevention group was significantly lower than that of normal saline solution group (P < 0.05) and prevention group (P < 0.01). The tumor formation time of Yifeixiaoliu granules prevention group was smaller than that of prevention treatment group (P < 0.05), and the carcinoma weight was greater than that of prevention treatment group (P < 0.05). Conclusion: The immunosenescence of mouse can be induced by D-galactose;can time-dependently delay the immunosenescence of mouse, by which Yifeixiaoliu Granules can prevent the occurrence and development of lung cancer, and can prolong survival period of the mice bearing lung tumor.