GPR120对脂多糖诱导的胰岛β细胞炎症损伤及TLR4/MyD88/NF-κB p65信号通路的影响-东南大学学报医学版

GPR120对脂多糖诱导的胰岛β细胞炎症损伤及TLR4/MyD88/NF-κB p65信号通路的影响

单位：秦皇岛市第一医院内分泌科, 河北秦皇岛 066000

关键词：G蛋白偶联受体120 脂多糖胰岛β细胞炎症 TLR4/MyD88/NF-κ B p65信号通路

分类号：R587

出版年·卷·期（页码）：2022·41·第一期（82-88）

摘要：

目的：探讨过表达G蛋白偶联受体120（GPR120）对脂多糖（LPS）诱导的胰岛β细胞炎症损伤及Toll样受体4（TLR4）/髓样分化因子（MyD88）/核转录因子-κB （NF-κB）信号通路的影响。方法：将MIN6细胞分为control组（细胞用无血清培养液进行培养）、LPS组（细胞用含10 μg·ml^-1 LPS的培养液进行培养）、LPS+control组（感染阴性对照慢病毒的细胞用10 μg·ml^-1 LPS培养液处理）、LPS+GPR120组（感染pcDNA3.1-GPR120慢病毒的细胞用10 μg·ml^-1 LPS培养液处理）4组。CCK-8法检测细胞增殖；Annexin-V-FITC/PI流式细胞术检测细胞凋亡；ELISA检测IL-6、IL-1β和TNF-α水平；蛋白质印迹法检测GPR120、TLR4、MyD88、NF-κB p65蛋白的表达。结果：LPS组GPR120蛋白相对表达量明显低于control组（P<0.05）；LPS+GPR120组GPR120蛋白相对表达量明显高于LPS+control组（P<0.05）。LPS组细胞增殖活力明显低于control组（P<0.001）；LPS+GPR120组细胞增殖活力明显高于LPS+control组（P<0.001）。LPS组细胞凋亡率明显高于control组（P<0.001）；LPS+GPR120组细胞凋亡率明显低于LPS+control组（P<0.001）。LPS组细胞上清液中IL-6、IL-1β和TNF-α水平均高于control组（均P<0.001）；LPS+GPR120组IL-6、IL-1β和TNF-α水平均低于LPS+control组（均P<0.001）。LPS组细胞TLR4、MyD88、NF-κB p65蛋白相对表达量均高于control组（均P<0.001），LPS+GPR120组TLR4、MyD88、NF-κB p65相对表达量均低于LPS+control组（均P<0.001）。结论：GPR120可能通过抑制TLR4/MyD88/NF-κB p65信号通路在LPS诱导的胰岛β细胞炎症损伤中发挥保护作用。

Objective: To investigate the effects of overexpression of G-protein coupled receptor 120(GPR120) on lipopolysaccharide(LPS) induced inflammation of islet beta cells and the Toll like receptor 4(TLR4)/myeloid differentiation factor(MyD88)/nuclear factor-κB(NF-κB) signaling pathway. Methods: MIN6 cells were divided into four groups:control group(cells were cultured in serum-free medium), LPS group(cells were cultured in medium containing 10 μg·ml^-1 LPS), LPS+control group(cells infected with negative control lentivirus were treated with 10 μg·ml^-1 LPS medium), and LPS+GPR120 group(cells infected with pcDNA3.1-GPR120 lentivirus were treated with 10 μg·ml^-1 LPS medium). CCK-8 method was used to detect cell proliferation; Annexin-V-FITC/PI flow cytometry was used to detect apoptosis; ELISA was used to detect IL-6, IL-1 β and TNF-α; Western blot was used to detect the expression of GPR120, TLR4, MyD88 and NF-κB p65. Results: The relative expression of GPR120 protein in LPS group was significantly lower than that in control group(P<0.05); the relative expression of GPR120 protein in LPS+GPR120 group was significantly higher than that in LPS+control group(P<0.05). The cell proliferation activity of LPS group was significantly lower than that of control group(P<0.001); the cell proliferation activity of LPS+GPR120 group was significantly higher than that of LPS+control group(P<0.001). The apoptosis rate of LPS group was significantly higher than that of control group(P<0.001); the apoptosis rate of LPS+GPR120 group was significantly lower than that of LPS+control group(P<0.001). The levels of IL-6, IL-1 β and TNF-α in the supernatant of LPS group were higher than those of control group(all P<0.001); the levels of IL-6, IL-1 β and TNF-α in LPS+GPR120 group were lower than those in LPS+control group(all P<0.001). The relative expression levels of TLR4, MyD88 and NF-κB p65 in LPS group were higher than those in control group(all P<0.001). The relative expression levels of TLR4, MyD88 and NF-κB p65 in LPS+GPR120 group were lower than those in LPS+control group(all P<0.001). Conclusion: GPR120 may play a protective role in LPS induced inflammatory injury of islet beta cells by inhibiting TLR4/MyD88/NF-κB p65 signaling pathway.

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