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黄芩水提液对3T3-L1脂肪细胞增殖、诱导分化及脂联素启动子荧光素酶活性的影响
引用本文:崔琳,路玲玲,李强,宰军华,刘卫红,王小晓. 黄芩水提液对3T3-L1脂肪细胞增殖、诱导分化及脂联素启动子荧光素酶活性的影响[J]. 世界科学技术-中医药现代化, 2015, 17(11): 2360-2366
作者姓名:崔琳  路玲玲  李强  宰军华  刘卫红  王小晓
作者单位:河南中医学院第一附属医院中心实验室 郑州 450000,聊城市中医院康复医学科 聊城 252000,河南中医学院第一附属医院中心实验室 郑州 450000;河南省病毒性疾病中医药防治重点实验室 郑州 450000,河南中医学院第一附属医院中心实验室 郑州 450000,河南中医学院第一附属医院中心实验室 郑州 450000,河南中医学院第一附属医院中心实验室 郑州 450000
基金项目:河南省教育厅自然科学研究资助计划项目(2010B360009):基于对脂肪细胞脂联素调节的减肥中药的初步筛选,负责人:李强。郑州市科技公关计划项目(0910SGYS33391-1):以脂肪细胞分泌因子为靶点的减肥中药的初步筛选,负责人:宰军华。
摘    要:
目的:本研究旨在观察黄芩水提液(Scutellaria Baicalensis Water Extract,SBWE)对3T3-L1前体细胞增殖、分化,对脂肪细胞因子脂联素表达以及脂联素(Adiponectin,ADP)启动子荧光素酶活性的影响,从分子生物学角度阐述SBWE降脂作用的可能机理。方法:通过体外培养3T3-L1细胞,采用MTT法检测SBWE对3T3-L1细胞增殖能力的影响;通过诱导脂肪细胞分化成为成熟脂肪细胞,观察SBWE对脂肪形成的影响;化学发光法检测脂联素启动子双荧光素酶报告基因活性;荧光定量PCR法检测脂联素mRNA(Adipoq)表达。结果: 与正常组相比,给予3T3-L1细胞0.01、0.1、1 mg?mL-1浓度的SBWE 24 h,可显著抑制细胞的增殖活性(P<0.05);0.1、1 mg?mL-1浓度的SBWE能够降低3T3-L1细胞分化为脂肪细胞的数量,并减少细胞内脂滴聚集,但无明显剂量依赖性;0.01、0.1 mg?mL-1浓度SBWE能显著提高脂联素基因启动子荧光素酶活性,与空载体比较差异有统计学意义(P<0.05);与正常组相比,给予3T3-L1细胞0.1 mg?mL-1 SBWE 24 h,诱导前后的脂肪细胞Adipoq表达均明显增加 (P<0.05)。结论: SBWE可有效抑制3T3-L1脂肪细胞的增殖、分化,同时增加脂联素基因表达,这可能是通过增强脂联素基因启动子荧光素酶活性实现,这些为黄芩水提液减肥的作用机制提供一定的基础。

关 键 词:黄芩 水提液 3T3-L1细胞 荧光定量PCR 细胞因子 脂联素 荧光素酶
收稿时间:2015-05-25
修稿时间:2015-11-05

The Effect of Scutellaria Baicalensis Water Extract on Proliferation, Cytokines mRNA Expressions and Promoter Activity of 3T3-L1 Cells
Cui Lin,Lu Lingling,Li Qiang,Zai Junhu,Liu Weihong and Wang Xiaoxiao. The Effect of Scutellaria Baicalensis Water Extract on Proliferation, Cytokines mRNA Expressions and Promoter Activity of 3T3-L1 Cells[J]. World Science and Technology—Modernization of Traditional Chinese Medicine and Materia Medica, 2015, 17(11): 2360-2366
Authors:Cui Lin  Lu Lingling  Li Qiang  Zai Junhu  Liu Weihong  Wang Xiaoxiao
Affiliation:The First Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou 450000, China,Department of Rehabilitation, Liaocheng Hospital of Traditional Chinese Medicine, Liaocheng 252000, China,The First Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou 450000, China; Henan Key Laboratory of Traditional Chinese Medicine Control Viral Diseases, Zhengzhou 450000, China,The First Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou 450000, China
Abstract:
The study was designed to measure the effect of S. baicalensis water extract (SBWE) on 3T3-L1 cells and its adiponectin (ADP) mRNA (Adipoq) and promoter luciferase activity. Cell survival rate was determined by MTT assay. The expression of Adipoq was measured by real-time PCR, while the luciferase report systems of Adipoq were used to transfer 3T3-L1 cells. The luciferase activities of the transferred cells were compared by luciferase assay. It was found that the mRNA expression of Adipoq was decreased in comparison with the control group. The luciferase activity showed a stronger ADP promoter activity in 3T3-L1 cells in SBWE treated group than that of control one. In conclusion, SBWE treatment improves the cykomine expression and luciferase reporter gene activity which will be an essential method for further studies of obesity therapy in traditional Chinese medicine.
Keywords:Scutellaria baicalensis   3T3-L1 cells   real-time PCR   cytokines   luciferase activity
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