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白藜芦醇抑制肠癌细胞焦亡的作用*
引用本文:任彩佩,张一楠,吴亚俐,都新新,崔香丽.白藜芦醇抑制肠癌细胞焦亡的作用*[J].中国应用生理学杂志,2022,38(4):326-331.
作者姓名:任彩佩  张一楠  吴亚俐  都新新  崔香丽
作者单位:山西医科大学生理学系, 太原 030001
基金项目:*山西省“1331工程”重点学科建设计划经费(1331KSC); 山西省自然科学基金(201801D121311); 细胞生理学教育部重点实验室运行经费(1C012019054)
摘    要:目的: 研究白藜芦醇(Res)对肠癌细胞焦亡的影响。方法: ①葡聚糖硫酸钠(DSS)诱发小鼠结肠癌(CRC)实验:30只C57BL/6小鼠随机分为正常对照组(Contro组),氧化偶氮甲烷(AOM)组,AOM/DSS组,AOM/DSS+Res组和Res组,每组6只,造模周期共70 d。第1周第1日对AOM组、AOM/DSS组和AOM/DSS+Res组小鼠AOM(10 mg/kg)腹腔注射一次,无菌水饮水,1% DSS水供AOM/DSS组和AOM/DSS+Res组饮用,对AOM/DSS+Res组和Res组小鼠灌胃给予Res(50 mg/kg),造模结束后,取小鼠结肠组织固定、包埋、切片; IHC和Western blot检测小鼠结肠组织NLRP3、Caspase-1、IL-18蛋白表达情况。②离体实验:HCT 116细胞给予Res(2.4 μg/L)以及转染miR-31,加Res实验分为4组,分别标记0 h、12 h、24 h、48 h组;细胞转染分组为5组,即control组、miR-31 mimic组、miR-31 mimic+Res组、miR-31inhibitor组、miR-31inhibitor+Res组,48 h后收集细胞,每组设置三个复孔,并通过Western blot检测细胞NLRP3、Caspase-1、GSDMD-N、IL-18和IL-1β蛋白表达情况。结果: 动物实验:与control组相比较,AOM/DSS组NLRP3、Caspase-1、IL-18蛋白表达显著升高(P<0.01),AOM/DSS+Res组NLRP3、Caspase-1、IL-18蛋白表达水平相较于AOM/DSS组有显著下降(P<0.01);细胞实验:与control组相比,miR-31 mimic组NLRP3(P<0.01)、GSDMD-N(P<0.05)、IL-18(P<0.01)蛋白表达显著升高, miR-31 inhibitor组NLRP3、GSDMD-N、IL-18蛋白表达显著降低(P<0.05)。结论: Res可通过细胞焦亡抑制结肠癌。

关 键 词:结肠癌  细胞焦亡  白藜芦醇  小鼠  炎症小体  HCT116细胞  细胞培养  
收稿时间:2022-03-03

Effects of resveratrol on inhibiting pyroptosis of intestinal cancer cells
REN Cai-pei,ZHANG Yi-nan,WU Ya-li,DU Xin-xin,CUI Xiang-li.Effects of resveratrol on inhibiting pyroptosis of intestinal cancer cells[J].Chinese Journal of Applied Physiology,2022,38(4):326-331.
Authors:REN Cai-pei  ZHANG Yi-nan  WU Ya-li  DU Xin-xin  CUI Xiang-li
Affiliation:Department of Physiology, Shanxi Medical University, Taiyuan 030001, China
Abstract:Objective: To study the effects of resveratrol (Res) on pyroptosis of colorectal cancer cells . Methods: ①The experiment of dextran sodium sulfate (DSS) induced colon cancer (CRC) in mice: 30 C57BL/6 mice were randomly divided into control group, Azoxymethane (AOM) group, AOM/DSS group, AOM/DSS+Res group and Res group, with 6 mice in each group, the modeling cycle was 70 days in total. Mice in AOM group, AOM/DSS group and AOM/DSS+Res group, at the first day of the first week, were intraperitoneally injected with AOM (10 mg/kg) once, and the ordinary chaw was replaced with high iron feed, and sterile water was given, 1% DSS water was given to AOM/DSS group and AOM/DSS+Res group. The mice in AOM/DSS+Res and Res groups were given resveratrol (50 mg/kg) by oral gavage, When the mold was finished, colon tissue of mice was fixed, embedded and sectionalized. The expressions of NLRP3, Caspase-1 and IL-18 in colon tissues of mice were detected by IHC and Western blot. ②In vitro experiment: HCT 116 cells were given Res (2.4 μg/L) and transfected with miR-31. The Res was divided into 4 groups and labeled with 0 h, 12 h, 24 h and 48 h respectively. The transfected cells were divided into 5 groups: Control group, miR-31 mimic group, miR-31 mimic + Res group, miR-31 inhibitor group, miR-31 inhibitor + Res group. The protein expressions of NLRP3, Caspase-1, GSDMD-N, IL-18 and IL-1β were detected by Western blot. Results: Animal experiments: Compared with control group, the protein expressions of NLRP3, Caspase-1 and IL-18 in AOM/DSS group were increased significantly (P<0.01). The protein expression levels of NLRP3, Caspase-1 and IL-18 in AOM/DSS+Res group were significantly lower than those in AOM/DSS group (P<0.01). Cell experiments: Compared with the control group, the protein expressions of NLRP3 (P<0.01), GSDMD-N (P<0.05) and IL-18 (P< 0.01) in miR-31 mimic group were increased significantly. The protein expressions of NLRP3, GSDMD-N and IL-18 in miR-31 inhibitor group were decreased significantly (P<0.05). Conclusion: Res inhibited the pyroptosis of colorectal cancer cells through pyroptosis.
Keywords:colon cancer  resveratrol  pyroptosis  mice  NLRP3  HCT116 cells  cell culture  
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