刺芒柄花苷对肺癌放疗诱发肠道炎症的影响及机制研究

The effects of ononin on radiation-induced intestinal inflammation in lung cancer and its underlying mechanisms

关键词:  刺芒柄花苷  肺癌  放疗  肠道炎症

Key words:  ononin  lung cancer  radiotherapy  intestinal inflammation

摘要:

目的 探究刺芒柄花苷对放疗诱发荷瘤小鼠肠道炎症的作用以及相关作用机制。方法 采用C57BL/6小鼠(18-22g,雄性)40只,随机分为空白组、肿瘤组、辐射组、刺芒柄花苷组。除空白组外,其余小鼠左侧腋窝皮下,接种小鼠肺癌Lewis细胞。辐射组和刺芒柄花苷组待肺部肿瘤长到可以触及皮下直径约3-6 mm肿块后进行X射线4Gy辐照其瘤体2min,建立肺癌荷瘤辐照小鼠模型。模型制备完成后,次日刺芒柄花苷组予以刺芒柄花苷连续腹腔注射3天。取材后,苏木素-伊红(HE)染色观察结肠组织病理学形态,ELISA检测IL-33、TNF-α、IL-1β、IL-6含量,免疫组化检测ST2L的表达,免疫荧光检测TRPA1的表达,蛋白免疫印迹法(Western blot,WB)检测ST2L、TRPA1、P-PLCγ、IP3R蛋白的表达。结果 与放疗后相比,刺芒柄花苷给药后结肠组织损伤明显改善(P＜0.01),肺组织中IL-33水平降低(P＜0.05),ST2L、TRPA1、P-PLCγ、IP3R蛋白表达显著下调(P＜0.01),血清中炎症因子TNF-α、IL-1β、IL-6水平降低(P＜0.01)。结论 刺芒柄花苷通过抑制放疗后肺部IL-33的表达进而抑制TRPA1通道的激活,从而缓解肺癌放疗诱发的肠道炎症。

Abstract:

ABSTRACT: OBJECTIVE The purpose of this study was to investigate the effect of cynandione A on radiation-induced intestinal inflammation in tumor-bearing mice and the underlying mechanisms. METHODS Forty C57BL/6 mice (18-22g, male) were randomly divided into blank group, tumor group, radiation group, and cynandione A group. Except for the normal group, the remaining mice were subcutaneously injected with Lewis lung cancer cells in the left axilla. The radiation group and the cynandione A group were irradiated with X-ray at a dose of 4Gy for 2 minutes after the lung tumors reached a diameter of approximately 3-6 mm. After the model was established, the cynandione A group was given intraperitoneal injections of cynandione A for 3 days. After sampling, hematoxylin-eosin (HE) staining was performed to observe the histopathological morphology of the colon tissue, ELISA was used to detect the levels of IL-33, TNF-α, IL-1β, and IL-6, immunohistochemistry was performed to detect the expression of ST2L, immunofluorescence was performed to detect the expression of TRPA1, and Western blot was used to detect the expression of ST2L, TRPA1, P-PLCγ, and IP3R proteins. RESULTS Compared with post-irradiation, cynandione A administration significantly improved colon tissue damage (P<0.01), decreased IL-33 levels in lung tissue (P<0.05), downregulated the expression of ST2L, TRPA1, P-PLCγ, and IP3R proteins (P<0.01), and reduced the levels of inflammatory factors TNF-α, IL-1β, and IL-6 in the serum (P<0.01). CONCLUSION Cynandione A alleviates radiation-induced intestinal inflammation in lung cancer by inhibiting the expression of IL-33 and subsequent activation of TRPA1 channels.