软肝饮对HBV转基因小鼠HBV-DNA、肝纤维化四项指标及炎症细胞因子表达的影响
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国家自然科学基金项目(81874451)


Effect of Ruangan Decoction on HBV-DNA, Four Markers of Liver Fibrosis, and Expression of Inflammatory Cytokines in HBV Transgenic Mice
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    摘要:

    目的 探讨中药软肝饮对乙型肝炎病毒(hepatitis B virus,HBV)转基因小鼠HBV-DNA、肝纤维化四项指标及炎症细胞因子表达的影响。方法 将50只雌雄各半的HBV转基因小鼠随机分为中药软肝饮治疗组(高剂量组、中剂量组、低剂量组)、恩替卡韦组、模型组和空白对照组,各组分别连续给药4周后取材。采用实时荧光定量PCR法检测外周血及肝组织中HBV-DNA表达水平,ELISA法检测血清中肝纤维化四项指标[层粘连蛋白(laminin,LN)、透明质酸(haluronic acid,HA)、Ⅳ型胶原(collage type Ⅳ,Ⅳ-C)、Ⅲ型前胶原(procollagen type Ⅲ,PCⅢ)]、炎症细胞因子[白细胞介素2(interleukin-2,IL-2)、白细胞介素4 (interleukin-4,IL-4)、白细胞介素10 (interleukin-10,IL-10)、干扰素γ(interferon γ,IFN-γ)]水平。结果 与模型组比较,软肝饮高、中、低剂量组和恩替卡韦组小鼠血清和肝组织中HBV-DNA表达水平明显降低,差异有统计学意义(P<0.05);软肝饮的作用呈明显的剂量依赖性(P<0.05)。与空白对照组比较,模型组小鼠血清LN、HA、Ⅳ-C、PCⅢ表达水平显著升高(P<0.05);与模型组比较,软肝饮高剂量组和恩替卡韦组LN、HA、Ⅳ-C、PCⅢ表达水平显著降低(P<0.05);软肝饮降低血清LN、HA、Ⅳ-C的作用呈剂量依赖性(P<0.05)。与空白对照组比较,模型组IL-2、IL-10、IFN-γ水平显著降低(P<0.05),IL-4水平显著升高(P<0.05);与模型组比较,软肝饮高、中剂量组和恩替卡韦组IL-4水平显著降低(P<0.05),IL-2、IL-10、IFN-γ水平显著升高(P<0.05);与软肝饮低剂量组比较,软肝饮高、中剂量组IL-4水平显著降低(P<0.05),IL-10水平显著升高(P<0.05)。结论 软肝饮治疗HBV转基因小鼠发挥抗病毒、抗纤维化作用机制可能与其调控炎症细胞因子水平有关。

    Abstract:

    Objective To investigate the effect of the traditional Chinese medicine Ruangan Decoction on hepatitis B virus (HBV)-DNA, four markers of liver fibrosis, and expression of inflammatory cytokines in HBV transgenic mice. Methods A total of 50 HBV transgenic mice (25 male mice and 25 female mice) were randomly divided into high-, middle-, and low-dose Ruangan Decoction treatment groups, entecavir group, model group, and blank control group, and related samples were collected after 4 consecutive weeks of administration. Quantitative real-time PCR was used to measure the level of HBV-DNA in peripheral blood and liver tissue; ELISA was used to measure the serum levels of the four markers of liver fibrosis [laminin (LN), hyaluronic acid (HA), collage type Ⅳ (Ⅳ-C), and procollagen type Ⅲ (PCⅢ)] and inflammatory cytokines [interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-10 (IL-10), and interferon-γ (IFN-γ)]. Results Compared with the model group, the high-, middle-, and low-dose Ruangan Decoction treatment groups and the entecavir group had a significant reduction in the level of HBV-DNA in serum and liver tissue (P<0.05), and the effect of Ruangan Decoction was dose-dependent (P<0.05). Compared with the blank control group, the model group had significant increases in the serum levels of LN, HA, Ⅳ-C, and PCⅢ (P<0.05); compared with the model group, the high-dose Ruangan Decoction treatment group and the entecavir group had significant reductions in the levels of LN, HA, Ⅳ-C, and PCⅢ (P<0.05); Ruangan decoction reduced the serum levels of LN, HA, and Ⅳ-C in a dose-dependent manner (P<0.05). Compared with the blank control group, the model group had significant reductions in the levels of IL-2, IL-10, and IFN-γ (P<0.05) and a significant increase in the level of IL-4 (P<0.05); compared with the model group, the high- and middle-dose Ruangan Decoction treatment groups and the entecavir group had a significant reduction in the level of IL-4 (P<0.05) and significant increases in the levels of IL-2, IL-10, and IFN-γ (P<0.05); compared with the low-dose Ruangan Decoction treatment group, the high- and middle-dose Ruangan Decoction treatment groups had a significant reduction in the level of IL-4 (P<0.05) and a significant increase in the level of IL-10 (P<0.05). Conclusion Ruangan Decoction exerts antiviral and anti-fibrotic effects on HBV transgenic mice possibly by regulating the levels of inflammatory cytokines.

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施 美,侯 勇,周 灏,刘丽丽,张先姚,张国梁.软肝饮对HBV转基因小鼠HBV-DNA、肝纤维化四项指标及炎症细胞因子表达的影响[J].安徽中医药大学学报,2021,40(4):74-78

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  • 在线发布日期: 2021-08-14