| 甘露聚糖结合凝集素(MBL)对3T3-L1前脂肪细胞成脂分化的调节作用及机制 |
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| 引用本文： | 杨泳慧,杨婧文,李志欣,孙彦威,杨易孟,张伟,王凡平,于莉莉,王明永.甘露聚糖结合凝集素(MBL)对3T3-L1前脂肪细胞成脂分化的调节作用及机制[J].中华微生物学和免疫学杂志,2020(2):122-128. |
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| 作者姓名： | 杨泳慧 杨婧文 李志欣 孙彦威 杨易孟 张伟 王凡平 于莉莉 王明永 |
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| 作者单位： | 新乡医学院医学检验学院;新乡医学院国际教育学院;新乡医学院基础医学院;河南省分子诊断与医学检验技术协同创新中心;新乡市免疫调控与分子诊断重点实验室 |
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| 基金项目： | 国家自然科学基金资助项目(81373120,31100645);河南省科技攻关计划项目(192102310303,202102310032);河南省高校科技创新团队支持计划(20IRTSTHN030);河南省教育厅科学技术研究重点项目(19A310002);河南省高等学校青年骨干教师培养计划(2018GGJS101)。 |
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| 摘 要： | 目的研究甘露聚糖结合凝集素(mannan-binding lectin,MBL)对3T3-L1前脂肪细胞成脂分化的调节及其机制。方法体外诱导3T3-L1前脂肪细胞成脂分化,同时给予不同浓度的MBL(0、1、10、20μg/ml)干预。CCK-8法检测细胞增殖能力变化,油红O染色和细胞内甘油三酯含量测定法分析脂质积累情况。Western blot及qRT-PCR检测脂肪细胞成脂分化相关因子PPARγ及C/EBPα的蛋白质及mRNA表达水平。Western blot分析脂肪合成调控信号分子Akt的表达及磷酸化。结果实验组各浓度MBL(0、1、10、20μg/ml)对3T3-L1前脂肪细胞增殖都无影响。3T3-L1前脂肪细胞诱导分化3 d,甘油三酯检测发现MBL处理组细胞内甘油三酯水平下降,并呈剂量依赖关系;油红O染色结果进一步显示,MBL处理组的脂滴数量显著减少,吸光度值也显著降低,同样呈现浓度依赖关系。Western blot及qRT-PCR检测结果证实,MBL处理组PPARγ和C/EBPα的蛋白质及mRNA表达水平均显著下降,呈剂量依赖性。在MBL干预下,Akt的磷酸化水平也明显下调。结论MBL通过Akt信号通路调控3T3-L1前脂肪细胞的成脂分化。
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| 关 键 词： | 甘露聚糖结合凝集素 3T3-L1前脂肪细胞 AKT 成脂分化 |
Regulation and mechanism of mannan-binding lectin(MBL)on adipogenic differentiation of 3T3-L1 preadipocytes |
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| Yang Yonghui,Yang Jingwen,Li Zhixin,Sun Yanwei,Yang Yimeng,Zhang Wei,Wang Fanping,Yu Lili,Wang Mingyong.Regulation and mechanism of mannan-binding lectin(MBL)on adipogenic differentiation of 3T3-L1 preadipocytes[J].Chinese Journal of Microbiology and Immunology,2020(2):122-128. |
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| Authors: | Yang Yonghui Yang Jingwen Li Zhixin Sun Yanwei Yang Yimeng Zhang Wei Wang Fanping Yu Lili Wang Mingyong |
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| Institution: | (School of Laboratory Medicine,Xinxiang Medical University,Xinxiang 453003,China;School of International Education,Xinxiang Medical University,Xinxiang 453003,China;School of Basic Medicine,Xinxiang Medical University,Xinxiang 453003,China;Henan Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine,Xinxiang 453003,China;Xinxiang Key Laboratory of Immunoregulation and Molecular Diagnostics,Xinxiang 453003,China) |
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| Abstract: | Objective To investigate the regulatory effects and mechanism of mannan-binding lectin(MBL)on adipogenic differentiation of 3T3-L1 preadipocytes.Methods 3T3-L1 preadipocytes were induced to differentiate into adipocytes in vitro,and stimulated with different concentrations of MBL(0,1,10,20μg/ml).Firstly,changes in cell proliferation ability were detected by CCK-8.Then lipid accumulation was analyzed by Oil red O staining and intracellular triglyceride content determination.Further,the expression of adipogenic differentiation-related factors PPARγand C/EBPαat protein and mRNA levels were detected by Western blot and qRT-PCR,respectively.Finally,Western blot was used to analyze the expression and phosphorylation of Akt,a signal molecule related to adipogenic differentiation.Results MBL at the concentrations of 0,1,10 and 20μg/ml had no effect on the proliferation of 3T3-L1 preadipocytes.The level of triglyceride in MBL treatment groups decreased in a dose-dependent manner on 3 d after 3T3-L1 preadipocyte differentiation.Results of Oil red O staining showed that the number of lipid droplets in MBL treatment groups reduced significantly,and the absorbance values also decreased significantly in a concentration-dependent manner.Western blot and qRT-PCR results showed that the expression of PPARγand C/EBPαat both protein and mRNA levels in MBL treatment groups decreased significantly in a dose-dependent manner,and the phosphorylation level of Akt was significantly down-regulated as well.Conclusions MBL regulates the adipogenic differentiation of 3T3-L1 preadipocytes via Akt signaling pathway. |
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| Keywords: | Mannan-binding lectin 3T3-L1 preadipocyte Akt Adipogenic differentiation |
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