论文部分内容阅读
通过研究半枝莲总黄酮(total flavonoids in Scutellaria barbata,TF-SB)对肿瘤细胞自噬以及炎症小体NLRP3的影响,探讨自噬与炎症小体NLRP3激活的关系,为进一步探讨TF-SB的抗肿瘤机制提供实验依据。本研究拟采用人工接种B16-F1细胞株致小鼠黑色素瘤模型,随机均分为模型对照组、阳性对照组(Rap,1.5 mg·kg-1)、TF-SB高、中、低(200,100,50 mg·kg-1)剂量组,另取健康C57BL/6J小黑鼠作为正常组以备血清检测对照,每组10只,每日给药1次,连续2周,末次药后30 min处理动物。采用HE染色检测肿瘤组织病理形态学改变;采用Western blot法检测肿瘤组织内LC3-Ⅱ/LC3-Ⅰ比值以及炎症小体NLRP3/caspase-1/IL-1β,IL-18的蛋白表达水平;采用ELISA法检测血清炎症因子IL-1β,IL-18含量。结果显示:模型组肿瘤细胞呈明显异型性及恶性增殖的特点,阳性对照组及TF-SB各剂量组肿瘤组织向周围侵犯情况明显减轻、肿瘤组织坏死范围明显增大、炎性反应明显减轻。阳性对照组及TF-SB各剂量组LC3-Ⅱ/LC3-Ⅰ比值明显增加,NLRP3/caspase-1/IL-1β,IL-18蛋白表达明显减少。模型组小鼠血清IL-1β,IL-18含量明显升高,与正常对照组比较有极显著性差异(P<0.001),阳性对照组及TF-SB各剂量组血清IL-1β,IL-18含量明显减少,上述指标各药物组与模型组比较均有显著性差异(P<0.05,P<0.01或P<0.001)。实验结果表明半枝莲总黄酮可通过抑制NLRP3炎症小体的表达、改变肿瘤生长微环境,从而产生抗肿瘤作用,其机制可能与其诱导肿瘤细胞发生自噬有关。
To study the effect of total flavonoids in Scutellaria barbata (TF-SB) on autophagy of tumor cells and the expression of NLRP3 in inflammatory cells, and to explore the relationship between autophagy and activation of inflammasome NLRP3. Anti-tumor mechanism to provide experimental evidence. In this study, the mouse melanoma model induced by artificial inoculation of B16-F1 cell line was used and randomly divided into model control group, positive control group (Rap, 1.5 mg · kg -1), high, medium and low TF- , 50 mg · kg-1), healthy C57BL / 6J black mice were taken as normal control group to prepare serum test control group, 10 rats in each group were given once a day for 2 weeks and 30 minutes after the last drug Handling animals. The histopathological changes of the tumor tissues were detected by HE staining. The expression of LC3-Ⅱ / LC3-Ⅰ and the expression of inflammatory factors NLRP3 / caspase-1 / IL-1β and IL-18 were detected by Western blot. Serum inflammatory cytokines IL-1β and IL-18 were detected by ELISA. The results showed that the tumor cells in the model group showed the characteristics of atypia and malignant proliferation. The tumor tissues of the positive control group and each dose of TF-SB were significantly reduced to the surrounding area, the scope of the tumor necrosis was significantly increased, and the inflammatory response was significantly alleviated. The ratio of LC3-Ⅱ / LC3-Ⅰ in the positive control group and TF-SB groups increased significantly, and the expression of NLRP3 / caspase-1 / IL-1β and IL-18 decreased significantly. The levels of IL-1β and IL-18 in the model group were significantly higher than those in the normal control group (P <0.001). The levels of IL-1β, IL- 18 significantly decreased, the above indicators of each drug group compared with the model group were significantly different (P <0.05, P <0.01 or P <0.001). Experimental results show that Scutellaria barbata total flavonoids can inhibit the expression of NLRP3 inflammasome, change the microenvironment of tumor growth, and thus have anti-tumor effect, and its mechanism may be related to its induction of tumor cell autophagy.