| MCHR1基因沉默对3T3-L1细胞诱导分化的影响 |
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| 引用本文： | 赵志武1,师磊1,陈显久2,王君实1,马敏1,燕炯1. MCHR1基因沉默对3T3-L1细胞诱导分化的影响[J]. 现代预防医学, 2015, 0(15): 2799-2802 |
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| 作者姓名： | 赵志武1 师磊1 陈显久2 王君实1 马敏1 燕炯1 |
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| 作者单位： | 1.山西医科大学公共卫生学院营养与食品卫生学教研室，山西 太原 030001；2.山西医科大学基础医学院生物化学与分子生物学教研室，山西 太原 030001 |
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| 摘 要： | 摘要:目的 运用RNA干扰技术沉默黑色素浓集激素受体1（Melanin-Concentrating Hormone Receptor 1，MCHR1）基因的表达，观察其对3T3-L1细胞诱导分化的影响，为肥胖症的基因治疗提供新思路。方法 用含绿色荧光蛋白的重组载体sh-MCHR1，通过脂质体法转染3T3-L1细胞。细胞诱导分化的同时用黑色素浓集激素（Melanin-Concentrating Hormone，MCH）干预，并在不同时点对脂滴进行油红O染色。采用RT-PCR以及Western blot分别检测过氧化物酶体增殖物激活受体γ（Peroxisome Proliferator-Activated Receptor γ，PPAR γ）、CCAAT/增强子结合蛋白-α（CCAAT/ Enhancer-Binding Protein α，C/EBPα）和脂肪酸结合蛋白2（adipocyte protein 2，ap2）mRNA和蛋白的表达。结果 重组载体sh-MCHR1成功转染；与阴性转染组相比，sh-MCHR1转染组d 10后，油红O 染液相对OD510值显著下降（P＜0.05）；PPARγ、C/EBPα和ap2 mRNA的表达量分别在d 6、d 8和d 8后显著下降（P＜0.05）；3者蛋白的表达量均在d 8后显著下降（P＜0.05）。结论 shRNA沉默MCHR1基因可减缓3T3-L1细胞诱导分化，其可能通过抑制PPARγ、C/EBPα和ap2的表达而实现。
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| 关 键 词： | 关键词:RNA干扰 MCHR1 3T3-L1细胞 细胞分化 |
Effect of silencing MCHR1 gene on the induction differentiation of 3T3-L1 adipocytes |
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| ZHAO Zhi-wu,SHI Lei,CHEN Xian-jiu,WANG Jun-shi,MA Min,YAN Jiong. Effect of silencing MCHR1 gene on the induction differentiation of 3T3-L1 adipocytes[J]. Modern Preventive Medicine, 2015, 0(15): 2799-2802 |
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| Authors: | ZHAO Zhi-wu SHI Lei CHEN Xian-jiu WANG Jun-shi MA Min YAN Jiong |
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| Affiliation: | Department of Nutrition and Food Hygiene, School of Public Health, Shanxi Medical University, Taiyuan, Shanxi 030001, China |
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| Abstract: | Abstract: Objective To block the melanin-concentrating hormone receptor 1 (MCHR1) expression with RNA interference and investigate the effect on the induction differentiation of 3T3-L1 preadipocytes. Methods The sh-MCHR1 was recombined with green fluorescence protein, after which 3T3-L1 preadipocytes was transfected by lipofectamine. Melanin-concentrating hormone (MCH) was applied at the same time to induce the differentiation of 3T3-L1 preadipocytes, and the lipid droplets were stained by red oil O at different time. The mRNA and protein expression of peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer-binding protein α (C/EBPα) and adipocyte protein 2 (ap2) were individually detected by RT-PCR and Western blots. Results Sh-MCHR1 recombinant vector was transfected into 3T3-L1 preadipocytes successfully. Compared to the negative control group, OD510 value dropped obviously after the 10th day in sh-MCHR1 group (P<0.05). Besides, the mRNA and protein expression of PPARγ, C/EBPα and ap2 were significantly inhibited on the 6th, 8th and 12th day (P<0.05). Conclusion Blocking MCHR1 gene could slow down the induction differentiation of 3T3-L1 preadipocytes by downregulating the expression of PPARγ, C/EBPα and ap2. |
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| Keywords: | Keywords: RNA interference MCHR1 3T3-L1 preadipocytes induction differentiation |
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