摘要
目的基于腺苷酸蛋白激酶(AMPK)-葡萄糖转运体4(GLUT4)轴探讨桑叶生物碱对妊娠糖尿病(GDM)小鼠糖耐量异常及胰岛素抵抗(IR)的作用机制。方法选取SPF级C57BL/6雌性小鼠60只和雄性小鼠30只,体重(20±3)g,6周龄。按照随机数字表法取50只处于发情期雌性小鼠,高糖脂饮食喂养6周后与25只雄性小鼠同笼过夜。受孕成功的45只雌性小鼠5 d后禁食12 h,腹腔注射35 mg/kg链脲佐菌素溶液,诱导GDM模型。造模后,25%葡萄糖1 ml灌胃,测其2 h餐后血糖(2h PBG),若7.8 mmol/L≤2h PBG<11.1 mmol/L,且持续1周即造模成功。将造模成功的40只雌小鼠依据随机数字表法分为模型组、桑叶生物碱低剂量组、桑叶生物碱高剂量组、阳性药物组,每组10只;余下10只未参与造模且妊娠成功的雌小鼠为对照组。桑叶生物碱低、高剂量组及阳性药物组分别尾静脉注射100、200、100 mg/kg药物溶液;对照组及模型组给予等体积无菌生理盐水。1次/d,共8周。给药第7周进行口服葡萄糖耐量试验,检测各组小鼠0.5、1、2 h血糖水平;给药第8周比较各组小鼠胰岛素抵抗指数稳态模型评估法(HOMA-IR),稳态模型胰岛β细胞功能指数(HOMA-β),C反应蛋白(CRP),肿瘤坏死因子-α(TNF-α),白细胞介素-6(IL-6),肝脏组织AMPK、GLUT4 m RNA及AMPK、磷酸化腺苷酸蛋白激酶(p-AMPK)、GLUT4蛋白表达水平。结果整体分析发现:对照组与模型组血糖时间、组间、交互作用比较,差异有统计学意义(P<0.05)。组内比较:对照组2 h血糖低于0.5、1 h,差异有统计学意义(P<0.05);模型组不同时间点血糖两两比较,差异有统计学意义(P<0.05)。组间比较:模型组0.5、1、2 h血糖高于对照组,差异有统计学意义(P<0.05)。整体分析发现:模型组与给药组血糖时间、组间、交互作用比较,差异有统计学意义(P<0.05)。组内比较:桑叶生物碱低、高剂量组不同时间点血糖两两比较,差异有统计学意义(P<0.05)。组间比较:桑叶生物碱低、高剂量组0.5、1、2 h血糖低于模型组;桑叶生物碱高剂量组0.5、1、2 h血糖低于桑叶生物碱低剂量组,差异有统计学意义(P<0.05)。模型组HOMA-IR,CRP、TNF-α、IL-6水平高于对照组;HOMA-β,肝脏组织GLUT4 m RNA及p-AMPK、GLUT4蛋白水平低于对照组,差异有统计学意义(P<0.05)。桑叶生物碱低、高剂量组HOMA-IR,CRP、TNF-α、IL-6水平低于模型组;HOMA-β,肝脏组织GLUT4 m RNA及p-AMPK、GLUT4蛋白水平高于模型组,差异有统计学意义(P<0.05)。桑叶生物碱高剂量组HOMA-IR,CRP、TNF-α、IL-6水平低于桑叶生物碱低剂量组,HOMA-β,肝脏组织GLUT4 m RNA及p-AMPK、GLUT4蛋白水平高于桑叶生物碱低剂量组,差异有统计学意义(P<0.05)。各组肝脏组织AMPK m RNA、蛋白水平比较,差异无统计学意义(P>0.05)。结论桑叶生物碱可改善GDM雌性小鼠血糖水平和炎症反应,减轻IR,促进葡萄糖代谢,且呈剂量依赖性。改善作用可能通过激活AMPK-GLUT4轴发挥作用。
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-pancreaticβ-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.
作者
乔艳华
白章莹
王俊芳
李晓敏
田莹
QIAO Yanhua;BAI Zhangying;WANG Junfang;LI Xiaomin;TIAN Ying(Department of Health,Handan Maternal and Child Health Hospital,Hebei Province,Handan 056000,China;Department of Obstetrics,Handan Maternal and Child Health Hospital,Hebei Province,Handan056000,China;Laboratory Medicine,Handan Maternal and Child Health Hospital,Hebei Province,Handan056000,China;Department of Pediatrics,Handan Maternal and Child Health Hospital,Hebei Province,Handan056000,China;Department of Obstetrics and Gynecology,Affiliated Hospital of Hebei University of Engineering,Hebei Province,Handan056001,China)
出处
《中国医药导报》
CAS
2023年第33期22-27,共6页
China Medical Herald
基金
河北省科技计划项目(19JH006572)。
关键词
妊娠糖尿病
桑叶生物碱
胰岛素抵抗
糖耐量异常
Gestational diabetes mellitus
Mulberry leaf alkaloids
Insulin resistance
Abnormal glucose tolerance
