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西地那非对Caco-2细胞增殖及对NCM460细胞炎症反应的影响

单静博, 郭沛艳, 陈特长, 汪云阳, 李晓祺, 周飒, 马文建

单静博, 郭沛艳, 陈特长, 汪云阳, 李晓祺, 周飒, 马文建. 西地那非对Caco-2细胞增殖及对NCM460细胞炎症反应的影响[J]. 中国药科大学学报, 2020, 51(1): 52-59. DOI: 10.11665/j.issn.1000-5048.20200109
引用本文: 单静博, 郭沛艳, 陈特长, 汪云阳, 李晓祺, 周飒, 马文建. 西地那非对Caco-2细胞增殖及对NCM460细胞炎症反应的影响[J]. 中国药科大学学报, 2020, 51(1): 52-59. DOI: 10.11665/j.issn.1000-5048.20200109
SHAN Jingbo, GUO Peiyan, CHEN Techang, WANG Yunyang, LI Xiaoqi, ZHOU Sa, MA Wenjian. Effects of sildenafil on the proliferation of Caco-2 cells and inflammatory response in NCM460 cells[J]. Journal of China Pharmaceutical University, 2020, 51(1): 52-59. DOI: 10.11665/j.issn.1000-5048.20200109
Citation: SHAN Jingbo, GUO Peiyan, CHEN Techang, WANG Yunyang, LI Xiaoqi, ZHOU Sa, MA Wenjian. Effects of sildenafil on the proliferation of Caco-2 cells and inflammatory response in NCM460 cells[J]. Journal of China Pharmaceutical University, 2020, 51(1): 52-59. DOI: 10.11665/j.issn.1000-5048.20200109

西地那非对Caco-2细胞增殖及对NCM460细胞炎症反应的影响

基金项目: 国家重点研发计划资助项目(No.2017YFD0400304);山东省重点研发计划资助项目(No.2018GSF121016)

Effects of sildenafil on the proliferation of Caco-2 cells and inflammatory response in NCM460 cells

  • 摘要: 为了探究西地那非对Caco-2细胞增殖的抑制作用及其对甲萘醌诱导的NCM460细胞炎症模型的抗炎作用,采用MTT法测定细胞的增殖能力;荧光探针法测定细胞内活性氧(ROS)及一氧化氮(NO)水平;Western blot检测Caco-2细胞中eNOS/ERK/JNK通路相关蛋白表达水平及NCM460细胞中相关炎症因子的蛋白表达水平;分光光度法测定西地那非对干酪乳杆菌及鼠李糖杆菌两种益生菌生长的影响。结果显示:西地那非可显著抑制Caco-2细胞的增殖,上调Caco-2细胞内eNOS、p-eNOS、p-JNK1/2及p-ERK1/2蛋白表达水平。西地那非与NG-L-硝基精氨酸甲酯(L-NAME)联用后,eNOS、p-eNOS、p-JNK1/2及p-ERK1/2蛋白表达水平较西地那非组明显降低。与甲萘醌组相比,西地那非可明显降低NCM460细胞内ROS水平,下调IL-6、IL-1β、p62、TNF-α蛋白表达水平。且高浓度西地那非对干酪乳杆菌及鼠李糖乳杆菌并无明显毒性。研究结果表明:西地那非在不破坏菌群稳态平衡的情况下,有效地抑制了肠道细胞炎症反应,通过抵抗外界条件引起的肠道细胞内的氧化应激反应,预防结直肠癌的发生。
    Abstract: To investigate the inhibitory effect of sildenafil on Caco-2 cell proliferation and its anti-inflammatory effect on menadione-induced NCM460 cell inflammation model, MTT assay was used to determine cell proliferation. Intracellular reactive oxygen species(ROS)and nitric oxide(NO)levels were detected by fluorescent probe. Western blot was used to detect the expression of eNOS/ERK/JNK pathway related proteins in Caco-2 cells and correlated inflammatory cytokines in NCM460 cells. The effect of sildenafil on the growth of two probiotics was determined by spectrophotometry. Results showed that sildenafil signi-ficantly inhibited the proliferation of Caco-2 cells and enhanced the expression levels of eNOS, p-eNOS, p-JNK1/2 and p-ERK1/2 proteins in Caco-2 cells; while after adding NG-nitro-L-arginine methyl ester(L-NAME), the expression levels of eNOS, p-eNOS, p-JNK1/2 and p-ERK1/2 proteins were significantly lower than those of the sildenafil group. Compared with the menadione group, sildenafil significantly reduced ROS levels in NCM460 cells and inhibited the expression levels of IL-6, IL-1β, p62, and TNF-α. Moreover, high concentrations of sildenafil had no obvious toxic effects on Lactobacillus casei and Lactobacillus rhamnosus. Thus, the results indicated that sildenafil could effectively inhibit the intestinal inflammatory response without affecting the balance of the intestinal flora, and prevent colorectal cancer by reducing the oxidative stress responses in the intestinal cells.
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  • 刊出日期:  2020-02-24

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