钠-葡萄糖共转运蛋白2抑制剂缓解精母细胞氧化应激改善2型糖尿病小鼠生精功能的研究  

Sodium-glucose co-transporter 2 inhibitor improves type 2 diabetes mellitus-induced testicular dysfunction by inhibiting oxidative stress of spermatocytes in mice

作　　者：夏利 张哲[2] 金滋润 姜雅菲 魏蕊[1] Xia Li;Zhang Zhe;Jin Zirun;Jiang Yafei;Wei Rui(Department of Endocrinology,State Key Laboratory of Female Fertility Promotion,Peking University Third Hospital,Beijing 100191,China;Department of Urology,Peking University Third Hospital,Beijing 100191,China)

机构地区：[1]北京大学第三医院内分泌科,女性生育力促进全国重点实验室,北京100191 [2]北京大学第三医院泌尿外科,北京100191

出　　处：《中华糖尿病杂志》2024年第8期889-897,共9页CHINESE JOURNAL OF DIABETES MELLITUS

基　　金：国家重点研发计划(2022YFC2702600)。

摘　　要：目的探讨钠-葡萄糖共转运蛋白2抑制剂(SGLT2i)对2型糖尿病(T2DM)小鼠睾丸生精功能的影响。方法将21只8周龄雄性db/db小鼠按随机数表法随机分为3组(每组7只),分别为db/db组[溶媒(水)干预]、达格列净(Dapa)组[达格列净(1 mg·kg^(-1)·d^(-1))]和卡格列净(Cana)组[卡格列净(10 mg·kg^(-1)·d^(-1))],每日灌胃,干预5周,另选5只同窝db/m小鼠作正常对照。监测所有小鼠的体重和血糖,干预结束后使用计算机辅助精子分析系统评价小鼠精液质量、HE染色评估睾丸组织形态学、脱氧核糖核苷酸末端转移酶介导的脱氧尿嘧啶核苷酸缺口末端标记(TUNEL)和免疫组化染色分析睾丸细胞凋亡情况。将小鼠精母细胞GC-2细胞系分为3组,分别为正常对照组(对照溶剂)、PA组(100μmol/L棕榈酸)、PA+Dapa组(100μmol/L棕榈酸联合30μmol/L达格列净),干预48 h,使用免疫印迹法、流式细胞术及试剂盒检测细胞氧化应激及凋亡情况。采用单因素方差分析或Brown-Forsythe/Welch进行组间比较。结果与db/db组相比,Dapa组和Cana组小鼠的空腹血糖、糖负荷后各时间点血糖水平均降低(均P<0.05);精子存活率和前向运动精子率均增加,曲线速度和平均路径速度均提升(均P<0.05);睾丸组织活化Caspase-3表达、TUNEL阳性生精小管比例和TUNEL阳性细胞比例均降低(均P<0.05)。GC-2细胞中,与PA组相比,PA+Dapa组细胞内活性氧水平、丙二醛含量、细胞凋亡率均降低,超氧化物歧化酶活性、B细胞淋巴瘤2(BCL2)表达均增加(均P<0.05)。结论SGLT2i改善T2DM小鼠生精功能障碍和精液质量下降,并通过减少睾丸组织氧化应激和凋亡发挥降糖非依赖的生殖系统直接保护作用。Objective To investigate the effect of sodium-glucose co-transporter 2 inhibitor(SGLT2i)on spermatogenesis and the quality of sperm in the mice with type 2 diabetes mellitus(T2DM).Methods Twenty-one 8-week-old male db/db mice were randomized into three groups(seven mice in each group)using a random number table method:db/db group(received vehicle),Dapa group[received dapagliflozin(1 mg·kg^(-1)·d^(-1))],and Cana group[received canagliflozin(10 mg·kg^(-1)·d^(-1))],orally daily for five weeks.Five littermate db/m mice were chosen as the normal control group.Body weight and the blood glucose levels were monitored in all mice.Sperm concentration and motility were analyzed using a computer-assisted semen analysis system.Testicular morphology was evaluated using HE staining,and testicular cell apoptosis was examined using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)and immunohistochemistry staining.GC-2 spermatogonia cells,were divided into three groups:normal control(vehicle),PA group(100μmol/L palmitic acid intervention),and PA+Dapa group(100μmol/L palmitic acid combined with 30μmol/L dapagliflozin intervention)and treated for 48 h.Oxidative stress levels and apoptosis-related indices were assessed using Western blotting,flow cytometry and test kits.One-way analysis of variance or Brown-Forsythe/Welch was used to compare between groups.Results Compared with db/db mice,dapagliflozin and canagliflozin significantly reduced fasting and postprandial blood glucose levels and the area under the glucose curve(all P<0.05).Both interventions increased sperm viability and progressive motility,as well as curve velocity and mean path velocity(all P<0.05).They also decreased activated caspase-3 expression,TUNEL-positive spermatogenic tubule ratio,and cell ratio(all P<0.05).In GC-2 cells,intervention with dapagliflozin reduced intracellular reactive oxygen species levels,malondialdehyde content,increased superoxide dismutase activity,decreased apoptosis rate,and increased B-cell lymphoma 2(BCL

关 键 词：糖尿病 2型 钠-葡萄糖共转运蛋白2抑制剂 精液质量 氧化应激 

分 类 号：R587.1[医药卫生—内分泌]