[1]邵敬芝,杜珊珊,张莉蓉,等.乳脂肪球表皮生长因子8（MFG-E8）对脂多糖诱导小胶质细胞炎症反应的抑制作用及其机制[J].眼科新进展,2021,41(10):910-914.[doi:10.13389/j.cnki.rao.2021.0191]
　SHAO Jingzhi,DU Shanshan,ZHANG Lirong,et al.Inhibiting effect and mechanism of MFG-E8 on lipopolysaccharide-induced microglial inflammatory response[J].Recent Advances in Ophthalmology,2021,41(10):910-914.[doi:10.13389/j.cnki.rao.2021.0191]
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乳脂肪球表皮生长因子8（MFG-E8）对脂多糖诱导小胶质细胞炎症反应的抑制作用及其机制/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
41卷
期数:
2021年10期
页码:
910-914
栏目:
实验研究
出版日期:
2021-10-05

文章信息/Info

Title:
Inhibiting effect and mechanism of MFG-E8 on lipopolysaccharide-induced microglial inflammatory response
作者:
邵敬芝; 杜珊珊; 张莉蓉; 张凤妍
450052　河南省郑州市，郑州大学第一附属医院眼科（邵敬芝，杜珊珊，张凤妍）；450000　河南省郑州市,郑州大学基础医学院药学部(张莉蓉)
Author(s):
SHAO Jingzhi1; DU Shanshan1; ZHANG Lirong2; ZHANG Fengyan1
1.Department of Ophthalmology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,Henan Province,China
2.Department of Pharmacology,School of Basic Medical Sciences,Zhengzhou University,Zhengzhou 450000,Henan Province,China
关键词:
乳脂肪球表皮生长因子8; 小胶质细胞; 炎症反应; 脂多糖
Keywords:
milk fat globule epidermal growth factor 8;  microglial cell;  inflammatory response;  lipopolysaccharide
分类号:
R774
DOI:
10.13389/j.cnki.rao.2021.0191
文献标志码:
A
摘要:
目的　探讨乳脂肪球表皮生长因子8（MFG-E8）对脂多糖（LPS）诱导小胶质细胞（BV-2细胞）炎症反应的抑制作用，并探讨可能作用机制。方法　CCK-8法检测0~200 mg·L-1 MFG-E8对细胞活力影响。建立LPS诱导BV-2细胞视网膜变性疾病体外模型，分为对照组、LPS组、LPS + MFG-E8组、LPS + LY294002组和LPS + PDTC组。倒置显微镜观察BV-2细胞的形态改变，ELISA检测肿瘤坏死因子-α（TNF-α）的表达变化，Western blot检测TNF-α和白细胞介素-6（IL-6）的蛋白表达变化，及PI3K-Akt和NF-κB相关通路蛋白表达变化。结果　0 mg·L-1、10 mg·L-1、50 mg·L-1、100 mg·L-1和200 mg·L-1的MFG-E8对BV-2细胞的活性无明显影响，细胞活力差异无统计学意义（P>0.05）；MFG-E8对LPS诱导的BV-2细胞形态变化具有一定的逆转作用；ELISA检测结果显示，LPS组BV-2细胞中TNF-α的含量较对照组明显升高（P<0.001），LPS + MFG-E8组BV-2细胞中TNF-α的含量较LPS组显著下降（P<0.001）。Western blot检测结果显示，LPS组BV-2细胞中TNF-α和IL-6蛋白相对表达量均显著高于对照组（均为P<0.05），LPS + MFG-E8组BV-2细胞中TNF-α和IL-6蛋白相对表达量均较LPS组明显降低（均为P<0.05）。LPS组BV-2细胞中NF-κB p-p65蛋白相对表达量明显高于对照组，p-PI3K、p-Akt蛋白相对表达量均明显低于对照组（均为P<0.001）；LPS + MFG-E8组BV-2细胞中NF-κB p-p65蛋白相对表达量较LPS组明显降低，p-PI3K、p-Akt蛋白相对表达量均较LPS组明显升高（均为P<0.001）。结论　MFG-E8可能是通过抑制NF-κB信号通路的激活，从而减轻LPS诱导的小胶质细胞炎症反应。
Abstract:
Objective　To investigate the inhibitory effect and mechanism of milk fat globule epidermal growth factor 8 (MFG-E8) on lipopolysaccharide (LPS)-induced microglia (BV-2 cell) inflammatory response.Methods　The effect of 0-200 mg·L-1 MFG-E8 on cell viability was detected by CCK-8 kits. An in-vitro model of LPS-induced BV-2 cell retinal degeneration was established, dividing into five groups: control group, LPS group, LPS + MFG-E8 group, LPS + LY294002 group and LPS + PDTC group. The morphological change of BV-2 cells was observed by inverted microscope. The expression change of tumor necrosis factor α (TNF-α) was detected by ELISA. The changes in protein expression levels of TNF-α, interleukin-6 (IL-6), PI3K-AKT and NF-κB related pathways associated protein were detected by Western blot. Results　Different concentrations of MFG-E8 (0 mg·L-1, 10 mg·L-1, 50 mg·L-1, 100 mg·L-1 and 200 mg·L-1) had no significant effect on the viability of BV-2 cells (P>0.05). MFG-E8 could reverse the morphological changes of BV-2 cells induced by LPS. ELISA results revealed that the concentration of TNF-α in BV-2 cells in the LPS group was significantly higher than that in control group (P<0.001). Compared with the LPS group, the concentration of TNF-α in BV-2 cells was significantly decreased in LPS + MFG-E8 group (P<0.001). Western blot showed that the relative protein expression levels of TNF-α and IL-6 in the LPS group were significantly higher than those in control group (both P<0.05). Compared with the LPS group, the relative protein expression levels of TNF-α and IL-6 were significantly decreased in LPS+ MFG-E8 group (both P<0.05). The relative protein expression level of NF-κB p-p65 in BV-2 cells in LPS group was significantly higher than that in the control group, while the relative protein expression levels of p-P13K and p-Akt were obviously lower than those in the control group (all P<0.001). The relative protein expression level of p-p65 in BV-2 cells in LPS + MFG-E8 group was lower than that in the LPS group, while the relative protein expression levels of p-P13K and p-Akt were obviously higher than those in the LPS group (all P<0.001). Conclusion　MFG-E8 may alleviate LPS-induced microglial inflammatory response by inhibiting the activation of NF-κB signaling pathway.

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备注/Memo

备注/Memo:
国家自然科学基金面上项目（编号：81970785）；河南省医学科技攻关联合共建项目（编号：2018020065）
更新日期/Last Update: 2021-10-05