1-脱氧野尻霉素减轻2型糖尿病小鼠的肝纤维化

摘要：目的 探讨 1-脱氧野霉素（DNJ）对糖尿病所致的肝纤维化的改善作用及其内在机制。方法 建立 2 型糖尿病小鼠（T2D）模 型，随机分为3组，5只/组：（1）对照组：腹膜内注射柠檬酸盐缓冲液（50 mg/kg）；（2）糖尿病组（DM）：采用高脂饮食饲养8周，静脉 注射 STZ（50 mg/kg），持续 3 d，空腹血糖浓度≥16.7 mmol/L；（3）DNJ 组：给予高脂饮食和 10%DNJ（200 mg·kg-1·d-1）饮水，持续 8 周。测定体质量（BW）、血糖、血清总胆固醇（TC）、甘油三酯（TG）、超氧化物歧化酶（SOD）。制作肝组织切片行苏木精伊红染色 （H&E）和天狼星红染色。此外，检测肝组织中胶原蛋白的溶解度。应用实时定量 PCR 法检测 MCP-1、TNF-α、IL-1β、TGF-β 1mRNA相对表达水平。Western-blot法测定α-SMA和Collagen2（ColA2）蛋白水平。将L929小鼠成纤维细胞用DNJ（10 μg/mL） 或 PBS 预处理 30 min，然后在含高浓度葡萄糖的 MEM 培养基培养 24 h。使用二氢乙锭（DHE）染色测定 ROS 的产生。结果 DNJ 可显著降低糖尿病小鼠血清中的血糖、TC、TG 和 SOD 水平（P<0.05），并显著减轻肝组织胶原交联程度，尤其以酶溶型胶原 （PSC）以及总胶原含量的减低为主要表现（P<0.05）。同时，DNJ 显著降低由糖尿病引起的促炎因子以及纤维化相关细胞因子 的高表达（P<0.05）。用 DNJ 预处理后培养的 L929 细胞在 DHE 染色中显示出较低的红色荧光强度（P<0.05）。结论 DNJ 作为 一种潜在的天然功能性营养物质，可以通过降血糖，抗炎以及抗氧化作用减轻 T2D 诱导的肝纤维化。 关键词：二型糖尿病；肝纤维化；1-脱氧野尻霉素；胶原交联；抗氧化应激
1-deoxynojirimycin alleviates liver fibrosis induced by type 2 diabetes in mice
HE Xiaoqiao2, SUN Zhiyuan3, MA Kaiyuan2, MEI Yingwu1 1Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou 450001, China; 2School of Basic Medical Sciences, Zhengzhou University, Zhengzhou 450001, China; 3School of Stomatology, Zhengzhou University, Zhengzhou 450052, China
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doi 10.12122/j.issn.1673-4254.2021.09.08
J South Med Univ, 2021, 41(9): 1342-1349
1-脱氧野尻霉素减轻 2 型糖尿病小鼠的肝纤维化
赫小乔 2，孙志远 3，马凯元 2，梅英武 1 1 郑州大学基础医学院生物化学与分子生物学系，河南 郑州 450001；2 郑州大学基础医学院，河南 郑州 450001；3郑州大学口腔医学院，河南 郑州 450052
ቤተ መጻሕፍቲ ባይዱ
Abstract: Objective To investigate the effect of 1-deoxynojirimycin (DNJ) for improving diabetic liver fibrosis and explore the underlying mechanism. Methods Mouse models of type 2 diabetes were established in 10 Kunming mice by high-fat diet feeding for 8 weeks and intraperitoneal injection of STZ, with 5 mice receiving intraperitoneal injection of citrate buffer solution with normal feeding as the control group. The mouse models were randomized into two groups (n=5) for further highfat feeding (model group) and additional treatment with 10% DNJ in drinking water (200 mg · kg-1 per day; DNJ group) for 8 weeks. The mice were monitored for changes in body weight (BW), blood glucose, serum total cholesterol (TC), triglyceride (TG) and superoxide dismutase (SOD) levels. The pathological changes in the liver tissue were observed using HE and Sirius Red staining, and the solubility of collagens in the liver tissues was determined. The expression levels of MCP-1, TNF-α, IL-1β and TGF-β1 mRNA were detected with real-time PCR, and the protein expressions of α-SMA and collagen2 (ColA2) were determined with Western blotting. In the in vitro experiment, mouse fibroblasts L929 cells were pretreated with DNJ (10 μg/ mL) or PBS for 30 min followed by culture in high-glucose medium for 24 h, and the level of ROS production was measured using dihydroethidium (DHE) staining. Results In the mouse model of type 2 diabetes, DNJ treatment significantly lowered serum level of glucose, TC, and TG (P<0.05) and increased serum SOD activity (P<0.05). DNJ obviously attenuated liver fibrosis in the diabetic mice, as shown by alleviated cross-linking of collagens and reduced contents of pepsin-solubilized collagen (PSC) and total collagen (P<0.05). DNJ treatment also significantly reduced the overexpression of the proinflammatory cytokines and fibrosis-related cytokines induced by diabetes (P<0.05). In L929 cells exposed to high glucose, pretreatment with DNJ significantly lowered the intensity of red fluorescence in DHE staining. Conclusion DNJ can attenuate type 2 diabetes-induced liver fibrosis in mice through its hypoglycemic, anti-inflammatory and anti-oxidative effects. Keywords: type2 diabetes; hepatic fibrosis; 1-deoxynojirimycin; collagen cross-linking; oxidative stress