| 肝豆灵通过lncRNA Meg3调控自噬减轻铜负荷Wilson病大鼠肝纤维化 |
| 投稿时间：2024-07-21 修订日期：2024-08-28 点此下载全文 |
| 引用本文：殷馨,汪瀚,孙兰婷,花代平,宣巧玉,杨文明.肝豆灵通过lncRNA Meg3调控自噬减轻铜负荷Wilson病大鼠肝纤维化[J].医学研究杂志,2025,54(1):37-42 |
| DOI：
10.11969/j.issn.1673-548X.2025.01.008 |
| 摘要点击次数: 109 |
| 全文下载次数: 98 |
| 作者 | 单位 | | 殷馨 | 安徽中医药大学第一临床医学院,合肥,230031 | | 汪瀚 | 安徽中医药大学第一附属医院神经内科,合肥,230031 | | 孙兰婷 | 安徽中医药大学第一附属医院神经内科,合肥,230031 | | 花代平 | 安徽中医药大学第一附属医院神经内科,合肥,230031 | | 宣巧玉 | 安徽中医药大学第一附属医院神经内科,合肥,230031 | | 杨文明 | 安徽中医药大学第一附属医院神经内科,合肥,230031 |
|
| 基金项目:国家自然科学基金区域创新发展联合基金资助项目(U22A20366)；安徽省中医药科技攻关专项(202303a07020004)；安徽省卫生健康科研项目(AHWJ2022b036)；安徽省自然科学基金资助项目(2208085MH266) |
|
| 中文摘要:目的 研究肝豆灵(gandouling, GDL)对铜负荷大鼠肝脏中长链非编码核糖核酸母系表达基因3(long noncoding RNA maternally expressed gene 3, lncRNA MEG3)和自噬的表达,并探讨其减轻肝纤维化的作用机制。方法 取10只SD大鼠作为正常组,另取50只分为模型组、青霉胺组、GDL低剂量组、GDL中剂量组、GDL高剂量组,每组10只。通过灌胃五水硫酸铜构建铜负荷Wilson病大鼠肝纤维化模型。采用苏木精-伊红(epithelial-mesenchymal transition, HE)、Masson染色观察肝脏病理形态；ELISA检测血清透明质酸(hyaluronic acid, HA)、层粘连蛋白(laminin, LN)、Ⅲ型前胶原肽(type Ⅲ procollagen, PC-Ⅲ)、Ⅳ型胶原(collagen Ⅳ, C-Ⅳ)水平；生化试剂盒检测谷丙转氨酶(alanine aminotransferase, ALT)、谷草转氨酶(aspartate aminotransferase, AST)活力；蛋白免疫印迹(Western blot)法检测α-平滑肌肌动蛋白(α-smooth muscle actin, α-SMA)、Ⅰ 型胶原(collagen type Ⅰ, Col-Ⅰ)蛋白表达；实时荧光定量聚合酶链式反应(real-time PCR,RT-qPCR)检测Beclin-1、lncRNA Meg3mRNA表达；免疫荧光法检测Beclin-1和微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3-Ⅱ,LC3-Ⅱ)表达。结果 与正常组比较,模型组大鼠肝组织出现胶原纤维沉积,AST、ALT、HA、LN、PC-Ⅲ、C-Ⅳ、α-SMA、Col-Ⅰ水平升高,lncRNA Meg3表达降低,自噬关键标志物Beclin-1和LC3-Ⅱ水平升高；给药GDL和青霉胺后,对比模型组,AST、LN、Col-Ⅰ、Beclin-1水平在 GDL低中高剂量组和青霉胺组都呈现降低趋势(P<0.01),C-Ⅳ、α-SMA水平在GDL低中高剂量组和青霉胺组中均下降,其中GDL中高剂量组降低显著(GDL低剂量组P<0.05,GDL中高剂量组P<0.01),而lncRNA Meg3降低表达被逆转,GDL中高剂量组和青霉胺组中lncRNA Meg3水平升高(P<0.01),病理切片结果显示,GDL干预后的大鼠肝纤维化程度减轻,GDL高剂量组效果明显。结论 肝豆灵可以减缓肝纤维化,其机制可能与lncRNA Meg3调控自噬表达有关。 |
| 中文关键词:肝豆灵 lncRNA Meg3 自噬 肝纤维化 |
| |
| GDL Alleviate Liver Fibrosis in Rats with Copper-Loaded Wilson′s Disease By Regulating Autophagy Through LncRNA Meg3. |
|
|
| Abstract:Objective To investigate the expression of long non-coding RNA maternally expressed gene 3 (lncRNA MEG3) and autophagy in the liver of copper-loaded rats treated with Gandouling (GDL) and to explore its mechanism of action in attenuating liver fibrosis. Methods Ten SD rats were taken as the normal group, and another 50 were divided into model groups, penicillamine group, GDL low dose group, GDL medium dose group, and GDL high dose group, with 10 rats in each group. The liver fibrosis model of rats with copper-loaded Wilson′s Disease was established by gavage of copper sulphate pentahydrate. HE and Masson′s staining were used to observe liver pathological morphology. ELISA was used to detect the levels of hyaluronic acid (HA), and laminin (LN), type Ⅲ procollagen (PC-Ⅲ), and type Ⅳ collagen (C-Ⅳ). A biochemical kit was used to detect the activity of alanine aminotransferase (ALT) and azelaic acid aminotransferase (AST). Western blot was used to detect the expression of alpha-smooth muscle actin (α-SMA) and collagen type Ⅰ (Col-Ⅰ). RT-qPCR was used to detect the expression of Beclin-1 and lncRNA Meg3mRNA. Immunofluorescence was used to detect the expression of Beclin-1 and LC3-Ⅱ. Results Compared with the normal group, rats in the model group showed collagen fiber deposition in liver tissue, increased levels of HA, LN, PC-Ⅲ, C-Ⅳ, α-SMA, Col-Ⅰ, liver injury, decreased expression of lncRNA Meg3, and increased levels of key markers of autophagy, Beclin-1 and microtubule-associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ). After administration of GDL and penicillamine, comparing to the model groups, the levels of AST, LN, Col-Ⅰ, and Beclin-1showed a trend of reduction in both GDL low, medium, and high dose groups and penicillamine group (P<0.01). The levels of C-Ⅳ, and α-SMA were decreased in the GDL medium, low, and high dose group and the penicillamine group, with a significant reduction in the GDL medium and high dose group (GDL low dose group:P<0.05, GDL middle-high dose group:P<0.01). Moreover, the reduced expression of lncRNA Meg3 was reversed, and the level of lncRNA Meg3 was increased in the GDL middle-high dose group and the penicillamine group (P<0.01). Pathological section results indicated that the degree of liver fibrosis was reduced in rats after GDL intervention, and the effect of GDL was obvious in the high dose group. Conclusion GDL alleviated liver fibrosis, and the mechanism may be related to the regulation of autophagy expression by lncRNA Meg3. |
| keywords:GDL LncRNA Meg3 Autophagy Hepatic fibrosis |
| 查看全文 查看/发表评论 下载PDF阅读器 |
|
|
|
