| 摘要: |
| 目的：观察抗肝纤维化细胞因子白介素10（IL-10）、肝细胞生长因子（HGF）和干扰素γ（IFN-γ）对含不同-509C>T基因型的转化生长因子β1（TGF-β1）基因上游启动子转录活性的影响，探讨细胞因子可能的抗肝纤维化机制。方法：以TGF-β1基因上游5′侧翼区启动子（-1328 ～ + 812）含有-509C>T 位点的片段作为靶序列,将其与氯霉素乙酰基转移酶（CAT）报告基因组成重组体phTGF2.14C、phTGF2.14T，脂质体法转染至人肝癌细胞系HepG2，应用IL-10（4 ng/ml）、HGF（10 ng/ml）和IFN-γ（20 ng/ml）作用于转染后HepG2细胞，ELISA法测定报告基因的表达。结果：转染phTGF2.14C的HepG2细胞报告基因活性明显高于转染phTGF2.14T者（P＜0.01）。IFN-γ对转染phTGF2.14C、phTGF2.14T的HepG2细胞TGF-β1启动子转录活性均具有明显的抑制作用（P＜0.05），HGF对转染phTGF2.14C HepG2细胞的TGF-β1启动子转录活性具有促进作用（P＜0.05），IL-10对两种情况下HepG2细胞TGF-β1启动子转录活性调控作用均不显著。结论：C等位基因可明显增强TGF-β1基因启动子转录活性；IFN-γ可能通过抑制TGF-β1基因启动子转录活性抑制肝纤维化，而HGF、IL-10的抗纤维化作用可能与此途径无关。 |
| 关键词: 多态性，单核苷酸 白细胞介素10 肝细胞生长因子 干扰素γ 转化生长因子β 转录，遗传 肝硬化 |
| DOI：10.3724/SP.J.1008.2007.01292 |
| 投稿时间：2007-09-18 |
| 基金项目:国家自然科学基金（30270605）. |
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| Regulatory effects of 3 kinds of antifibrotic cytokines on activity of transforming growth factor-β1 gene promoter |
| HOU Li-na, GAO Chun-fang*, ZHAO Yun-peng, SUN Xiao-juan, CHEN Jian-dong，JIANG Xiao-jiong |
| (Department of Laboratory Medicine, Eastern Hepatobiliary Hospital, Second Military Medical University,Shanghai 200438, China) |
| Abstract: |
| Objective：To study the regulatory effects of antifibrotic cytokines, interleukin 10 (IL-10), hepatocyte growth factor (HGF), and interferon-gamma (IFN-γ) on activity of transforming growth factor-β1(TGF-β1 ) gene promoter, so as to assess the antifibrotic mechanism of cytokines. Methods: Sequence -1328-+812 of TGF-β1 gene, which contains the -509 C>T polymorphism, was selected as putative promoter. The recombinant constructions containing -1328-+812 of TGF-β1 gene and CAT reporter gene (phTGF2.14T, phTGF2.14C) were constructed and transfected into HepG2 cells with liposomal transfection method, then the transfected HepG2 cells were treated with IL-10（4 ng/ml）, HGF（10 ng/ml） or IFN-γ（20 ng/ml）. Reporter gene activity was analyzed by ELISA. Results: Reporter gene activity in cells transfected with phTGF2.14C was significantly higher than those transfected with phTGF2.14T (P＜0.01). IFN-γ significantly inhibited the reporter gene activity in HepG2 cells transfected with phTGF2.14C or phTGF2.14T(P＜0.05); HGF significantly increased the reporter gene activity in cells transfected with phTGF2.14C (P＜0.05). IL-10 had no effects on the activities of cells transfected with phTGF2.14C or phTGF2.14T.Conclusion: C allele at -509 can increase the promoter activity of TGF-β1 gene in HepG2 cells. The antifibrotic effect of IFN-γ might be related to its inhibitory effect on the putative promoter activity of TGF-β1 gene; the antifibrotic effects of HGF and IL-10 may not be through regulation of TGF-beta1 gene transcription. |
| Key words: polymorphism,single nucleotide interleukin-10 hepatocyte growth factor interferon-gamma transforming growth factor beta transcription,genetic liver cirrhosis |
