目的 探究维生素D（vitamin D ,VD）缺乏加重糖尿病（diabetes mellitus,DM）小鼠心肌损伤的机制。方法 雄性6 w龄db/m小鼠和db/db小鼠按照体重和空腹血糖随机分为4组,每组8只（n=8）：对照组（Con）,VD缺乏对照组（def-Con）,模型组（DM）,VD缺乏模型组（def-DM）,喂养17 w,def-Con和def-DM组给予VD缺乏饲料。检测小鼠血糖、血脂、肌酸激酶同工酶（creatine kinase isoenzyme MB,CK-MB）水平,HE染色检测心肌组织病理变化,超声心动图评估小鼠心功能,Western blot检测心脏NADPH氧化酶4（NADPH oxidase 4,NOX4）-核苷酸结合寡聚化结构域样受体蛋白3（Nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3）炎性小体通路及焦亡相关蛋白表达水平。结果 与Con组相比,DM组小鼠体重、进食量显著增加,心脏重量/体重比值显著降低,血糖、血脂水平显著升高（P<0.05）;与DM组相比,def-DM组小鼠体重、进食量进一步增加,心脏重量/体重之比进一步降低,血脂水平进一步升高（P<0.05）,血糖水平无显著差异（P>0.05）。与Con组相比,DM组小鼠心肌细胞明显肥大,心脏LVEF、LVFS显著降低（P<0.05）;与DM组相比,def-DM组小鼠心肌细胞进一步肥大,心脏LVEF、LVFS进一步降低（P<0.05）。与Con组相比,DM组小鼠心脏NOX4/NLRP3炎性小体和焦亡相关蛋白表达水平显著升高（P<0.05）;与DM组相比,def-DM组小鼠心脏NOX4/NLRP3炎性小体和焦亡相关蛋白表达水平进一步升高（P<0.05）。结论 VD缺乏可能通过激活NOX4/NLRP3炎性小体通路加重糖尿病小鼠心肌焦亡损伤。
Abstract
Objective To investigate the mechanisms through which vitamin D (VD) deficiency exacerbates myocardial injury in diabetic mice. Methods Male 6-week-old db/m and db/db mice were randomly divided into four groups based on body weight and fasting blood glucose levels (n=8 per group): control group (Con), vitamin D deficiency control group (def-Con), model group (DM), and vitamin D deficiency model group (def-DM). All mice were fed for a duration of 17 weeks, with the def-Con and def-DM groups mice receiving a vitamin D-deficient diet. Serum glucose, lipid profiles, and CK-MB levels were measured. Myocardial histopathological changes were detected using HE staining. Cardiac function was assessed via echocardiography. Protein expression levels associated with the NOX4/NLRP3 inflammasome pathway and pyroptosis in cardiac tissue were determined by Western blotting. Results Compared with the Con group, DM mice exhibited significant increases in body weight and food intake, alongside notable reductions in the heart weight/body weight ratio, and elevated serum glucose and lipid levels (P<0.05). Moreover, compared to the DM group, the def-DM group displayed further increases in body weight and food intake, additional reductions in the heart weight/body weight ratio, and higher serum lipid levels (P<0.05), with no significant disparity in serum glucose levels (P>0.05). Cardiomyocyte hypertrophy was significantly pronounced in the DM group when compared to the Con group, accompanied by substantial reductions in cardiac LVEF and LVFS (P<0.05). The def-DM group displayed exacerbated cardiomyocyte hypertrophy and decreased cardiac LVEF and LVFS when compared to the DM group (P<0.05). Notably, the expression levels of NOX4/NLRP3 inflammasome and pyroptosis-related proteins were significantly elevated in the cardiac tissues of mice in the DM group compared to the Con group (P<0.05). Furthermore, the def-DM group presented further elevation in the expression levels of these proteins in cardiac tissues compared to the DM group (P<0.05). Conclusion Vitamin D deficiency may aggravate myocardial pyroptosis injury in diabetic mice by activating the NOX4/NLRP3 inflammasome pathway.
关键词
维生素D缺乏 /
糖尿病小鼠心肌损伤 /
NLRP3炎性小体 /
焦亡
Key words
vitamin D deficiency /
myocardial injury in diabetic mice /
NLRP3 inflammasome /
pyroptosis
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基金
河北省教育厅科学研究项目（No.QN2022016）; 河北省中央引导地方科技发展资金项目（No.246Z7760G）; 河北医科大学自然科学领域青年科技人才支持计划（No.CYQD2023001）
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