HAmLET诱导小鼠前脂肪细胞3T3-L1成脂分化  

HAmLET Induces Adipogenic Differentiation of Mouse Preadipocytes 3T3-L1 Cells

作　　者：张轶博 钟悦 曾瑞霞 Zhang Yibo;Zhong Yue;Zeng Ruixia(Department of Pathogenic Biology,Jinzhou Medical University,Jinzhou 121000 China;Department of Ophthalmology,Central Hospital of Yingkou Development Zone,Yingkou 115007 China;Department of Human Anatomy,Jinzhou Medical University,Jinzhou 121000 China)

机构地区：[1]锦州医科大学病原生物学教研室,辽宁锦州121000 [2]营口经济开发区中心医院眼科,辽宁营口115007 [3]锦州医科大学人体解剖学教研室,辽宁锦州121000

出　　处：《锦州医科大学学报》2022年第4期14-19,共6页Journal of Jinzhou Medical University

基　　金：辽宁省科技厅自然基金项目,项目编号:2019-MS-143;辽宁省科技厅自然科学博士启动基金项目,项目编号:2020-BS-253。

摘　　要：目的肿瘤细胞致死性人α-乳清蛋白(humanα-lactalbumin made lethal to tumor cells,HAmLET)是一种可以选择性杀伤各种肿瘤细胞的人α-乳清蛋白与油酸的脂蛋白复合物,但其对未分化细胞的作用研究较少,本研究探讨HAmLET对小鼠前脂肪细胞3T3-L1成脂分化的作用及其机制。方法使用不同浓度HAmLET作用3T3-L1细胞和肿瘤细胞U87MG 24 h或7 d后通过CCk8实验检测细胞活性,选择合适浓度HAmLET与胰岛素、IBMX、地塞米松三联法诱导3T3-L1细胞成脂分化,12 d后通过油红O染色观察脂肪细胞内脂质聚集及脂滴大小、异丙醇脂质萃取检测脂质含量、Western Blot检测PPARγ的蛋白表达。结果50、100、200、500μg/mL HAmLET作用3T3-L1细胞24 h后,3T3-L1的细胞活性均显著高于U87MG细胞(P<0.01),而3T3-L1的细胞活性在100和200μg/mL之间显著降低(P<0.01)。100μg/mL和200μg/mL HAmLET作用3T3-L1细胞7 d后细胞存活率没有明显变化,而100μg/mL组镜下观察到少数3T3-L1细胞体积增大、变圆,胞质内可见脂滴。油红O染色,相对脂质含量及PPARγ蛋白检测结果显示100μg/mL HAmLET诱导3T3-L1细胞分化12 d后脂质含量水平和PPARγ蛋白表达水平与经典三联法诱导组差异无统计学意义(P>0.05)另外,HAmLET可显著提高诱导细胞中。结论使用能杀伤肿瘤细胞U87MG细胞浓度的100μg/mL HAmLET可通过激活3T3-L1细胞中PPARγ蛋白表达诱导细胞分化成脂。Objective Humanα-lactalbumin made lethal to tumor cells(HAmLET)is a lipoprotein of humanα-lactalbumin and oleic acid that is able to selectively kill various tumor cells.Until now,there are few studies on undifferentiated cells.This study was to investigate the effect of HAmLET on the adipogenic differentiation of mouse preadipocytes 3T3-L1 and its mechanism.Methods Cell viability of 3T3-L1 cells and tumor cells treated with U87MG for 24 h or 7 d were detected by CCk8 assay.After 12 days,lipid accumulation and lipid droplet size in adipocytes were observed by oil red O staining,lipid content was detected by isopropanol lipid extraction,and protein expression of PPARγwas detected by Western Blot.Results After treated with 50,100,200,500μg/mL HAmLET,the cell activity of 3T3-L1 cells was significantly higher than that of U87MG cells(P<0.01),while the cell activity of 3T3-L1 cells decreased significantly between 100 and 200μg/mL(P<0.01).The survival rate of 3T3-L1 cells treated with 100μg/mL and 200μg/mL HAmLET did not change significantly after 7 days,while the volume of a few 3T3-L1 cells increased and became round and lipid droplets were observed in the cytoplasm of 100μg/mL group.The results of oil red O staining,relative lipid content and PPARγprotein detection showed that there were no significant differences in lipid content and PPARγprotein expression between classical triad induction group and the group in which 100μg/mL HamLET induced 3T3-L1 cells differentiation for 12 days(P>0.05).Furthermore,HAmLET could significantly enhance the induction of cells.Conclusion The use of HAmLET at a concentration of 100μg/mL that can kill tumor cells U87MG cells is able to induce cell differentiation into adipocytes by activating the expression of PPARγprotein in 3T3-L1 cells.

关 键 词：HAMLET 3T3-L1前脂肪细胞 脂肪分化 PPARΓ OA 

分 类 号：R363.1[医药卫生—病理学]