[关键词]
[摘要]
目的:探讨白术(Atractylodes macrocephala)水提物抑制胃癌SGC-7901细胞活性的潜在机制。方法:分别使用蒸馏水(对照)和白术水提物(白术治疗组)灌胃SD大鼠后,采集静脉血后分离其血清、过滤并分别命名为对照组血清(CON-S)和白术组血清(AM-S)。将胃癌SGC7901细胞分为对照组、10%AM-S组和20%AM-S组,其中两个AM-S组细胞分别在相应浓度的AM-S血清中培养24 h,对照组细胞用正常培养基培养相同时间,收取SGC7901细胞和上清液用于进一步分析。使用MTT法检测各组细胞活力,通过商业试剂盒测定乳酸脱氢酶(LDH)、丙二醛(MDA)和超氧化物歧化酶(SOD)的水平,采用ELISA试剂盒检测各组细胞中IL-6和TNF-α的含量,采用WB法评估各组细胞中PI3K-Akt-NF-κB信号通路相关蛋白的表达。结果:10%AM-S组和20%AM-S组的SGC7901胃癌细胞增殖活力相较于对照组分别降低48.9%和53.25%(P<0.05或P<0.01);胃癌细胞上清液中,相较于对照组,10%AM-S组和20%AM-S组LDH水平分别升高29.25%和123%、SOD活性分别升高18%和54.60%、MDA水平分别降低27.8%和40.0%,IL-6水平分别降低15%和17.5%、TNF-α水平分别降低29.71%和40.16%(P<0.05或P<0.01)。相较于对照组,AM-S组中PI3K-Akt-NF-κB信号相关蛋白的水平显著下降(P<0.05或P<0.01)。结论:白术水提物可以通过抑制癌细胞增殖活力、促进凋亡、抑制肿瘤微环境中的促炎因子分泌以及改变细胞内的氧化应激水平等方式抑制胃癌,其机制可能是通过抑制PI3K-Akt-NF-κB通路来实现这些抗癌作用的。
[Key word]
[Abstract]
Objective: To investigate the potential mechanism of Atractylodes macrocephala aqueous extract inhibiting gastric cancer SGC7901 cells. Methods: Gastric perfusion was performed on SD rats using distilled water and Atractylodes macrocephala aqueous extract respectively. Venous blood was collected and insolated to get serum, which was then filtered and named as control group serum (CON-S) and Atractylodes macrocephala group serum (AM-S), respectively. SGC7901 cells were divided into control group, 10% AM-S group and 20% AM-S group. The two AM-S group cells were cultured in AM-S serum of corresponding density for 24 h while the control group cells were cultured with normal medium for the same time. The SGC7901 cells and supernatant were collected for further analysis. MTT assay was used to detect cell viability. Lactate dehydrogenase (LDH), malondialdehyde (MDA) and superoxide dismutase (SOD) were determined by commercial kits. Enzyme-linked immuno sorbent assay (ELISA) kits were applied to detect thecontents of interleukin IL-6 and tumor necrosis factor TNF- α in the cells of all the groups. The expressions of phosphatidylinositol 3-kinase PI3K-Akt-NF- κB signaling pathway-related proteins were evaluated by western blot. Results: The proliferative activity of SGC7901 gastric cancer cells in 10% AM-S group and 20% AM-S group decreased by 48.9% and 53.25% respectively compared with the control group (P<0.05 or P<0.01). In the supernatant of gastric cancer cells, compared with the control group, the LDH level of 10% AM-S group and 20% AM-S group increased by 29.25% and 123%; the SOD activity increased by 18% and 54.60%; the MDA leveldecreased by 27.8% and 40.0%; the IL-6 level decreased by 15% and 17.5%, and the TNF-α αlevel decreased by 29.71% and 40.16% respectively (P<0.05 or P<0.01). Compared with the control group, the levels of PI3K-Akt-NF-κB signaling pathway-related protein in AM-S group were significantly reduced (P<0.05 or P<0.01). Conclusion: Atractylodes macrocephala aqueous extract can inhibit gastric cancer by inhibiting the proliferation activity of cancer cells, promoting apoptosis, inhibiting the production of pro-inflammatory factors in tumor microenvironment and changing the level of intracellular oxidative stress, the mechanism of which might be that these anti-cancerous effects are achieved by inhibiting PI3K-Akt-NF-κB signaling pathway.
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[基金项目]
江苏省老年健康科研面上项目(No. LKM2023026)