内毒素耐受对小鼠腹腔巨噬细胞分泌MMP⁃1、MMP⁃7及细胞迁移能力的影响
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国家自然科学基金(81771075);江苏省自然科学基金(BK20161565);江苏省高校优势学科建设工程资助项目(2018?87)


Effects of endotoxin tolerance on the secretion of MMP⁃1,MMP⁃7 in mice peritoneal macrophages and the migration of L929 cells
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    摘要:

    目的:观察牙龈卟啉单胞菌(Porphyromonas gingivalis,P.gingivalis)脂多糖(lipopolysaccharide,LPS)诱导的耐受对小鼠腹腔巨噬细胞分泌基质金属蛋白酶(matrix metalloproteinase,MMP)-1和MMP-7的影响,以及耐受巨噬细胞对小鼠成纤维细胞L929迁移能力的影响。方法:采用1 μg/mL P.gingivalis LPS重复刺激小鼠腹腔巨噬细胞,构建内毒素耐受模型,并以1 μg/mL大肠杆菌(Escherichia coli,E.coli)LPS作为阳性对照。收集细胞条件培养液上清,采用ELISA技术检测MMP-1和MMP-7表达水平。观察耐受巨噬细胞条件培养液对L929细胞划痕创伤构建后细胞迁移能力的影响。结果:P. gingivalis LPS重复刺激后,巨噬细胞MMP-1分泌水平较单次刺激组降低(P < 0.05),MMP-7分泌水平无明显变化(P > 0.05)。P. gingivalis LPS诱导的耐受巨噬细胞培养液上清刺激12、24 h后,L929细胞划痕修复面积大于单次刺激组培养液上清刺激后(P < 0.05)。结论:P. gingivalis LPS诱导的耐受能抑制小鼠腹腔巨噬细胞分泌MMP-1,促进L929细胞迁移。

    Abstract:

    Objective:To observe the effects of endotoxin tolerance induced by Porphyromonas gingivalis(P.gingivalis)on the secretion of metalloproteinase(MMP)-1 and MMP-7 in peritoneal macrophages of mice and the wound-healing abilities of L929 cells. Methods:Endotoxin tolerance in murine peritoneal macrophages was triggered by repeated 1 μg/mL P.gingivalis lipopolysaccharide(LPS)stimulations,and 1 μg/mL Escherichia coli(E.coli) LPS was served as positive control. Supernatants from tolerized macrophages were collected and levels of MMP-1 and MMP-7 were determined by ELISA. In addition,L929 cells were stimulated by supernatants from tolerized macrophages and their wound-healing abilities were evaluated by repair areas after scratch. Results:Secretion of MMP-1 in macrophages decreased significantly after repeated stimulation by P.gingivalis LPS compared with that treated by single P.gingivalis LPS stimulation(P < 0.05),while there were no differences in MMP-7 levels between the two groups(P > 0.05). Repaired areas after scratch(12 h,24 h)in L929 cells cultured with supernatants from P.gingivalis LPS-tolerized macrophages were bigger than those from P.gingivalis LPS non-tolerized macrophages(P < 0.05). Conclusion:Endotoxin tolerance triggered by P.gingivalis LPS contributes to the decreased secretion of MMP-1 in murine peritoneal macrophages and the enhanced migration of L929 cells.

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汪 悦,顾建雨,朱奕名,张尧尧,孙 颖.内毒素耐受对小鼠腹腔巨噬细胞分泌MMP⁃1、MMP⁃7及细胞迁移能力的影响[J].南京医科大学学报(自然科学版),2020,(1):21-25

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  • 收稿日期:2019-06-14
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  • 在线发布日期: 2020-02-09
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