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| Study of the mechanism of mulberry leaf alkaloids on abnormal glucose tolerance and insulin resistance in mice with gestational diabetes mellitus based on AMPK-GLUT4 axis |
| QIAO Yanhua1 BAI Zhangying2▲ WANG Junfang3 LI Xiaomin4 TIAN Ying5 |
1.Department of Health, Handan Maternal and Child Health Hospital, Hebei Province, Handan 056000, China;
2.Department of Obstetrics, Handan Maternal and Child Health Hospital, Hebei Province, Handan 056000, China;
3.Laboratory Medicine, Handan Maternal and Child Health Hospital, Hebei Province, Handan 056000, China;
4.Department of Pediatrics, Handan Maternal and Child Health Hospital, Hebei Province, Handan 056000, China;
5.Department of Obstetrics and Gynecology, Affiliated Hospital of Hebei University of Engineering, Hebei Province, Handan 056001, China |
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Abstract Objective To explore the mechanism of mulberry leaf alkaloids on abnormal glucose tolerance and insulin resistance (IR) in mice with gestational diabetes mellitus (GDM) based on the AMP-activated protein kinase (AMPK)- glucose transporter 4 (GLUT4) axis. Methods Sixty SPF C57BL/6 female mice and 30 male mice with body weight of (20±3) g and age of six weeks were selected. According to the random number table method, 50 female mice in estrus were selected, after being fed a high-glycemic diet for six weeks, 25 male mice were caged overnight. After five days of fasting for 12 h, 45 female mice with successful pregnancy were injected with 35 mg/kg streptozotocin solution peritoneally to induce GDM model. After the modeling, 25% glucose was given to the stomach with 1 ml, and the 2-hour postpranpranal blood glucose (2hPBG) was measured, if 7.8 mmol/L≤2hPBG＜11.1 mmol/L, and the modeling lasted for one week, the modeling was successful. According to random number table method, 40 female mice were divided into model group, mulberry leaf alkaloids low-dose group, mulberry leaf alkaloids high-dose group, and positive drug group, with ten mice in each group; the remaining ten female mice who did not participate in modeling and had successful pregnancy were selected as control group. Mulberry leaf alkaloids low-, high-dose groups and positive drug groups were injected with 100, 200 mg/kg, and 100 mg/kg drug solution through tail vein, respectively; control group, and model group were given equal volume sterile saline. Once a day for eight weeks. Oral glucose tolerance test was performed at the 7th week of administration, and blood glucose levels were detected at 0.5, 1 h, and 2 h; the expression levels of homeostasis model assessment for insulin resistance (HOMA-IR), homeostasis model assessment-pancreatic β-cell function (HOMA-β), C-reactive protein (CRP), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), AMPK and GLUT4 mRNA in liver tissues, and AMPK, phosphorylated adenylate protein kinase (p-AMPK), and GLUT4 protein in liver tissues of mice in all groups were compared at the 8th week of administration. Results Overall analysis showed that there were significant differences in blood glucose time, intergroup, and interaction between control group and model group (P＜0.05). Intra-group comparison: the level of blood glucose at 2 h in control group was lower than that at 0.5 and 1 h, and the differences were statistically significant (P＜0.05); the pairwise comparison of the levels of blood glucose at different time points in model group had statistical significance (P＜0.05). Comparison between groups: the levels of blood glucose at 0.5, 1 h, and 2 h in model group were higher than those in control group, and the differences were statistically significant (P＜0.05). Overall analysis showed that there were significant differences in blood glucose time, intergroup, and interaction between model group and drug administration group (P＜0.05). Intra-group comparison: the pairwise comparison of the levels of blood glucose in mulberry leaf alkaloids low-, high-dose groups at different time points showed statistical significance (P＜0.05). Comparison between groups: the levels of blood glucose in mulberry leaf alkaloids low-, high-dose groups at 0.5, 1 h, and 2 h were lower than those in model group; the levels of blood glucose at 0.5, 1 h, and 2 h in mulberry leaf alkaloids high-dose group were lower than those in mulberry leaf alkaloids low-dose group, and the differences were statistically significant (P＜0.05). HOMA-IR and the levels of CRP, TNF-α, IL-6 in model group were higher than those in control group; HOMA-β, GLUT4 mRNA， and the levels of p-AMPK and GLUT4 protein in liver tissues were lower than those in control group, and the differences were statistically significant (P＜0.05). HOMA-IR and the levels of CRP, TNF-α, IL-6 in mulberry leaf alkaloids low-, high-dose groups were lower than those in model group; HOMA-β, GLUT4 mRNA, and the levels of p-AMPK and GLUT4 protein in liver tissue weres higher than those in control group, and the differences were statistically significant (P＜0.05). HOMA-IR and the levels of CRP, TNF-α, IL-6 of mulberry leaf alkaloids high-dose group were lower than those of mulberry leaf alkaloids low-dose group; HOMA-β, GLUT4 mRNA，and the levels of p-AMPK and GLUT4 protein in liver tissues were higher than that of mulberry leaf alkaloids low-dose group, and the differences were statistically significant (P＜0.05). There were no significant differences in AMPK mRNA and protein in liver tissues among all groups (P＞0.05). Conclusion Mulberry leaf alkaloids can improve blood glucose level and inflammatory response, reduce IR and promote glucose metabolism in a dose-dependent manner in female GDM mice. The improvement may be effected by activation of the AMPK-GLUT4 axis.
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[1] Plows JF，Stanley JL，Baker PN，et al. The pathophysiology of gestational diabetes mellitus [J]. Int J Mol Sci，2018，19（11）：3342.
[2] 张立雯，季涛，宿树兰，等.桑叶黄酮类和生物碱类成分在正常和糖尿病大鼠体内的药代动力学研究[J].中国中药杂志，2017，42（21）：4218-4225.
[3] 石婧婧.桑叶活性成分及药理作用研究进展[J].生物化工，2023，9（1）：179-183.
[4] 郝麒麟，彭晓蝶，杨敏，等.桑叶生物碱对高脂饮食小鼠的抗氧化作用[J].现代食品科技，2019， 35（6）：39-47.
[5] 阴新负，彭效明，李翠清，等.桑叶生物碱和多糖的分离纯化研究进展[J].中医药导报，2016，22（14）：108-110.
[6] 杨忠敏，沈以红，王祖文，等.桑叶生物碱对D-半乳糖诱导氧化损伤模塑小鼠肾脏的改善作用[J].食品科学，2020， 41（19）：198-203.
[7] 王祖文，沈以红，黄先智，等.桑叶生物碱对肝纤维化小鼠抗氧化能力及炎症因子水平的影响[J].食品科学，2020， 41（17）：202-207.
[8] Gvazava IG，Kosykh AV，Rogovaya OS，et al. A simplified streptozotocin-induced diabetes model in nude mice [J]. Acta Naturae，2020，12（4）：98-104.
[9] 徐文婷，汤梦雨，王彩红，等.葛根素对妊娠期糖尿病大鼠糖脂代谢作用的研究[J].南京中医药大学学报，2019， 35（4）：448-452.
[10] 应巧，何斐，张伟，等.基于IRS-1信号通路探讨虎杖甙对2型糖尿病大鼠胰岛素抵抗的改善作用[J].新中医，2020，52（17）：4-8.
[11] White SL，Pasupathy D，Sattar N，et al. Metabolic profiling of gestational diabetes in obese women during pregnancy [J]. Diabetologia，2017，60（10）：1903-1912.
[12] Skorzynska-dziduszko KE，Kimber-trojnar ■，Patro-mal- ysza J，et al. An interplay between obesity and inflammation in gestational diabetes mellitus [J]. Curr Pharm Biotechnol，2016，17（7）：603-613.
[13] 中华医学会糖尿病学分会.中国2型糖尿病防治指南（2020年版）[J].中华糖尿病杂志，2021，13（4）：315-409.
[14] Gagne-Ouellet V，Houde AA，Guay SP，et al. Placental lipoprotein lipase DNA methylation alterations are associated with gestational diabetes and body composition at 5 years of age [J]. Epigenetics，2017，12（8）：616-625.
[15] Zambon M，Mando C，Lissoni A，et al. Inflammatory and oxidative responses in pregnancies with obesity and periodontal disease [J]. Reprod Sci，2018，25（10）：1474-1484.
[16] Sproston NR，Ashworth JJ. Role of C-reactive protein at sites of inflammation and infection [J]. Front Immunol，2018， 9：754.
[17] Garcia D，Shaw RJ. AMPK：mechanisms of cellular energy sensing and restoration of metabolic balance [J]. Mol Cell，2017，66（6）：789-800.
[18] 刘凡铭，王琪，钱昱臻，等.腺苷酸活化蛋白激酶在糖脂代谢调控中的研究进展[J].生物工程学报，2019，35（6）： 1021-1028.
[19] Herzig S，Shaw RJ. AMPK：guardian of metabolism and mitochondrial homeostasis [J]. Nat Rev Mol Cell Biol，2018，9（2）：121-135.
[20] 张珊，徐梦珠，郝娟，等.基于磷脂酰肌醇-3-激酶/蛋白激酶B/葡萄糖转运蛋白4通路的中药治疗糖尿病新进展[J].基层中药，2022， 1（4）：82-88.
[21] Kim J，Yang G，Kim Y，et al. AMPK activators：mechanisms ofaction and physiological activities [J]. Exp Mol Med，2016， 48（4）：e224.
[22] 王晓萌，杨鹏，李晓晨，等.基于AMPK信号通路改善T2DM 胰岛素抵抗中药有效成分研究进展[J/OL].中国实验方剂学杂志:1-10[2023-10-24].https://doi.org/10.13422/j.cnki.syfjx.20231204.
[23] Cai S，Sun W，Fan Y，et al. Effect of mulberry leaf （Folium Mori） on insulin resistance via IRS-1/PI3K/Glut-4 signalling pathway in type 2 diabetes mellitus rats [J]. Pharm Biol，2016，54（11）：2685-2691.
[24] Bae UJ，Jung ES，Jung SJ，et al. Mulberry leaf extract displays antidiabetic activity in db/db mice via Akt and AMP- activated protein kinase phosphorylation [J]. Food Nutr Res，2018，22：62.
[25] 高迎春，岳颖，周珺，等.桑叶提取物通过调节肝线粒体融合分裂相关因子改善2型糖尿病大鼠的胰岛素抵抗[J].中国药理学与毒理学杂志，2019，33（9）：743. |
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