梓醇调控糖酵解改善KK-Ay自发性糖尿病小鼠睾丸病变的作用及机制  

Improvement effects and its mechanism of catalpol on testicular lesions in KK-Ay spontaneous diabetic mice by regulating glycolysis

作　　者：陈玉萍 束安梅 许惠琴[2] 蒋鸣 朱逸晖 CHEN Yuping;SHU Anmei;XU Huiqin;JIANG Ming;ZHU Yihui(Center of Basic Medicine,Jiangsu Vocational College of Medicine,Jiangsu Yancheng 224005,China;College of Pharmacy,Nanjing University of Traditional Chinese Medicine,Nanjing 210023,China)

机构地区：[1]江苏医药职业学院基础医学部,江苏盐城224005 [2]南京中医药大学药学院,南京210023

出　　处：《中国药房》2023年第7期784-789,795,共7页China Pharmacy

基　　金：国家自然科学基金资助项目(No.81903879);江苏高校“青蓝工程”中青年学术带头人项目(No.苏教师函2021〔11〕号);江苏医药职业学院教育科学研究立项课题(No.Y202101)。

摘　　要：目的基于晚期糖基化终末产物(AGEs)及其受体(RAGE)介导的糖酵解过程,研究梓醇对KK-Ay自发性糖尿病小鼠睾丸病变的改善作用及机制。方法以高脂饲料喂养的KK-Ay自发性糖尿病小鼠作为糖尿病模型,随机分为模型组、梓醇组(100 mg/kg)、氨基胍组(AGEs抑制剂,100 mg/kg)和FPS-ZM1组(RAGE抑制剂,1 mg/kg),并将同期饲养的C57BL/6J小鼠作为正常组,每组6只。梓醇组和氨基胍组小鼠灌胃相应药物,正常组和模型组小鼠灌胃等量生理盐水,FPS-ZM1组小鼠以1 mL/g腹腔注射相应药物,连续8周。末次给药后,检测小鼠体质量、空腹血糖和24 h摄食量、饮水量、尿量以及睾丸脏器系数、精子活力;观察小鼠睾丸组织病理形态学和超微结构;检测小鼠睾丸组织中还原型谷胱甘肽(GSH)、超氧化物歧化酶(SOD)、乳酸脱氢酶(LDH)和糖代谢产物水平,并进行通路的富集分析;检测小鼠血清和睾丸组织中AGEs水平,睾丸组织中RAGE、B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)表达水平以及糖酵解关键限速酶[己糖激酶(HK)、磷酸果糖激酶(PFK)、丙酮酸激酶(PK)、LDH]相应mRNA表达水平。结果经梓醇干预后,KK-Ay自发性糖尿病小鼠的一般体征、睾丸脏器系数及精子活力均显著改善(P<0.05或P<0.01);生精小管中各层生精细胞的形态以及超微结构均有所改善;睾丸组织中GSH、SOD、LDH水平,糖代谢产物果糖-1,6-二磷酸、3-磷酸甘油酸、3-磷酸甘油醛、乳酸、丙酮酸水平,以及HK、PFK、PK、LDH mRNA表达水平均显著升高(P<0.05或P<0.01);血清和睾丸组织中AGEs水平、睾丸组织中RAGE蛋白表达水平和Bax/Bcl-2均显著降低(P<0.05或P<0.01)。经氨基胍和FPS-ZM1干预后,小鼠上述大部分指标均显著改善(P<0.05或P<0.01)。结论梓醇对KK-Ay自发性糖尿病小鼠睾丸病变有明显的改善作用,其作用机制可能与上调AGEs/RAGE信号通路介导的糖酵解有关。OBJECTIVE To study the improvement effects and its mechanism of catalpol on testicular lesions in KK-Ay spontaneous diabetic mice on the basis of glycolysis process mediated by advanced glycation end products(AGEs)and their receptors(RAGE).METHODS KK-Ay spontaneous diabetic mice fed with high-fat diet were used as diabetic model,and then randomly divided into model group,catalpol group(100 mg/kg),aminoguanidine group(AGEs inhibitor,100 mg/kg)and FPSZM1 group(RAGE inhibitor,1 mg/kg),and C57BL/6J mice fed in the same period were set as normal group,with 6 mice in each group.The catalpol group and aminoguanidine group mice were given relevant medicine intragastrically,normal group and model group mice were given constant volume of normal saline intragastrically,and FPS-ZM1 group mice were given relevant medicine 1 mL/g intraperitoneally,for consecutive 8 weeks.After the last administration,the body mass,fasting blood glucose,24-hour food intake,water consumption,urine volume,testicular organ coefficient,and sperm motility of the mice were measured;pathological morphology and ultrastructural structure of testicular tissue were observed;the levels of reduced glutathione(GSH),superoxide dismutase(SOD),lactate dehydrogenase(LDH)and sugar metabolites in testicular tissue of mice were detected;pathway enrichment analysis was performed;the level of AGEs in serum and testicular tissue,protein expressions of RAGE,B-cell lymphoma-2(Bcl-2)and Bcl-2-associated X protein(Bax),and mRNA expressions of key rate-limiting enzymes[hexokinase(HK),phosphofructose kinase(PFK),pyruvate kinase(PK),LDH]in testicular tissue were all detected.RESULTS Catalpol could significantly improve the general symptoms,testicular organ coefficients and motility of sperm in KK-Ay spontaneous diabetic mice(P<0.05 or P<0.01).The morphology and ultrastructure of spermatogenic cells in each layer of the seminiferous tubules were all improved.The levels of GSH,SOD and LDH in testicular tissue,the levels of the metabolic product glucose fructose-1,6-diphosphate

关 键 词：梓醇 糖酵解 糖尿病 睾丸病变 晚期糖基化终末产物 晚期糖基化终末产物受体 

分 类 号：R965[医药卫生—药理学]