التفاصيل البيبلوغرافية
| العنوان: |
miR-144-3p 参与高蛋氨酸饮食诱导 Cbs+/- 小鼠的肝细胞自噬. (Chinese) |
| العنوان البديل: |
Involvement of miR-144-3p in Cbs+/- mouse hepatocyte autophagy induced by high-methionine diet. (English) |
| المؤلفون: |
盛思琪, 谢 琳, 赵翔宇, 姜怡邓, 吴 凯, 熊建团, 杨安宁, 郝银菊, 焦 运 |
| المصدر: |
Chinese Journal of Tissue Engineering Research / Zhongguo Zuzhi Gongcheng Yanjiu; 3/18/2024, Vol. 28 Issue 8, p1289-1294, 6p |
| مصطلحات موضوعية: |
FATTY liver, AUTOIMMUNE hepatitis, PEARSON correlation (Statistics), CELL survival, GENE knockout, ASPARTATE aminotransferase |
| الملخص (بالإنجليزية): |
BACKGROUND: High-methionine diet can cause liver injury in Cbs+/- mice, and hyperhomocystinemia is related to the occurrence and progression of various liver-related diseases, such as hepatic steatosis, autoimmune hepatitis, and alcoholic fatty liver disease. MicroRNAs (miRNAs) are involved in various cellular processes including cell survival, differentiation and autophagy, which are of great significance. OBJECTIVE: To investigate the critical role of miR-144-3p on Cbs+/- mouse hepatocyte autophagy induced by high methionine die. METHODS: (1) Ten male cystathione-β-synthase normal (Cbs+/+) mice and another 10 male mice with single gene knockout (Cbs+/-) of similar body mass, 4 weeks of age, were fed a high-methionine diet and executed after 12 weeks to take liver tissue. (2) Human hepatocytes (HL-7702) were cultured in vitro and divided into control [0 μmol/L homocysteine (Hcy)], Hcy (100 μmol/L Hcy), mimic-NC (transfected with mimic-NC), mimic-NC + Hcy (mimic-NC transfecton+100 μmol/L Hcy), miR-144-3p mimic (transfected with miR-144-3p mimic), and miR-144-3p mimic + Hcy (miR-144-3p mimic transfection+100 μ mol/L Hcy), inhibitor-NC (transfected with inhibitor-NC), inhibitor-NC + Hcy (inhibitor-NC transfection + 100 μmol/L Hcy), miR-144-3p inhibitor (transfected with miR-144-3p inhibitor), and miR-144-3p inhibitor + Hcy (miR-144-3p inhibitor transfection + 100 μmol/L Hcy). Quantitative real-time PCR was used to detect the expression of miR144-3p in liver tissue and hepatocytes. After transfection of miR-144-3p mimic or inhibitor, quantitative real-time PCR and western blot were used to detect the transfection efficiency of miR-144-3p and its effect on the expression of autophagy-related proteins LC3B and p62. The levels of alanine transferase and aspartate aminotransferase in hepatocyte supernatants were determined by enzyme linked immunosorbent assay. The correlation between the expression of miR-144-3 in hepatocyte and the levels of alanine transferase and aspartate aminotransferase in hepatocyte supernatants were analyzed by Pearson correlation analysis. RESULTS AND CONCLUSION: Compared with the Cbs+/+ group and control group, the expression of miR-144-3p in the liver tissue of the Cbs+/- group and in hepatocytes of the Hcy group was decreased (P < 0.01). The expression of LC3B-II/I was decreased in hepatocyte after transfection of miR-144-3p mimic, while the protein expression of p62 was increased (P < 0.01). The opposite results were obtained after transfection of miR-144-3p inhibitor (P < 0.01). Compared with the mimic-NC group, the levels of alanine transferase and aspartate aminotransferase were decreased in the miR-144-3p mimic group (P < 0.01), while the opposite results were obtained in the inhibitor-NC group (P < 0.01). The expression of miR-144-3p in hepatocytes was negatively correlated with the levels of alanine transferase (P < 0.01, r=-0.887 6) and aspartate aminotransferase (P < 0.01, r=-0.829 9) in the supernatant of hepatocytes. To conclude, Hcy promotes hepatocyte autophagy by inhibiting the expression of miR-144-3p, which subsequently aggravates liver injury. [ABSTRACT FROM AUTHOR] |
| Abstract (Chinese): |
背景：高蛋氨酸饮食可导致Cbs+/- 小鼠发生肝损伤, 高同型半胱氨酸血症与肝脂肪变性、自身免疫性肝炎、酒精性脂肪肝等多种肝脏相关 疾病的发生和进展有关。微小RNA (miRNAs)参与细胞存活、分化和细胞自噬等各种细胞过程, 具有重要意义。 目的：探讨miR-144-3p在高蛋氨酸饮食诱导Cbs+/- 小鼠肝细胞自噬中的关键作用。 方法：①选取4周龄体质量相近的雄性胱硫醚β-合成酶基因正常(Cbs+/+)小鼠和单基因敲除(Cbs+/- )小鼠各10只, 均饲以高蛋氨酸饮食, 12 周后处死, 留取肝脏组织。②体外培养人源肝细胞(HL-7702), 分为对照组(0 μmol/L同型半胱氨酸)、同型半胱氨酸组(100 μmol/L同型半 胱氨酸)、mimic-NC组(转染mimic-NC)、mimic-NC+同型半胱氨酸组(转染mimic-NC+100 μmol/L同型半胱氨酸)、miR-144-3p mimic组(转染 miR-144-3p mimic)、miR-144-3p mimic+同型半胱氨酸组(转染miR-144-3p mimic+100 μmol/L同型半胱氨酸)、inhibitor-NC组(转染inhibitor-NC)、 inhibitor-NC+同型半胱氨酸组(转染inhibitor-NC+100 μmol/L同型半胱氨酸)、miR-144-3p inhibitor组(转染miR-144-3p inhibitor)、miR-144-3p inhibitor+同型半胱氨酸组(转染miR-144-3p inhibitor+100 μmol/L同型半胱氨酸)。采用荧光定量PCR检测肝组织和肝细胞中miR-144-3p的表达 水平；转染miR-144-3p模拟物或抑制剂后, 采用荧光定量PCR和Western blot分别检测miR-144-3p的转染效率及其对LC3B和p62蛋白表达的影 响；酶联免疫法检测肝细胞上清液中丙氨酸氨基转移酶和门冬氨酸氨基转移酶的表达情况；Pearson相关性分析肝细胞miR-144-3p表达与肝 细胞上清液中丙氨酸氨基转移酶和门冬氨酸氨基转移酶含量的相关性。 结果与结论：①与Cbs+/+组比较, Cbs+/- 组小鼠肝组织和同型半胱氨酸组肝细胞中miR-144-3p的表达水平降低(P < 0.01)；②转染miR-144-3p 模拟物后, 与mimic-NC比较, miR-144-3p mimic组中LC3B-Ⅱ/Ⅰ蛋白的表达水平降低, p62蛋白的表达水平升高(P < 0.01)；转染miR-144-3p inhibitor后, 得到相反的结果(P < 0.01)；③与mimic-NC组相比, miR-144-3p mimic组肝细胞上清液中丙氨酸氨基转移酶和门冬氨酸氨基转移 酶的含量降低(P < 0.01)；与inhibitor-NC组比较, 得到相反的结果(P < 0.01)；④肝细胞中miR-144-3p的表达与肝细胞上清液中丙氨酸氨基转 移酶(P < 0.01, r=-0.887 6)和门冬氨酸氨基转移酶(P < 0.01, r=-0.829 9)的含量呈负相关；⑤结果表明, 同型半胱氨酸通过抑制miR-144-3p 的表达促进肝细胞的自噬, 进而加重肝损伤。 [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |
