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脐带间充质干细胞在糖尿病大鼠中的自噬调节及肾脏保护作用

Autophagy regulation and renal protection of umbilical cord mesenchymal stem cells in diabetic rats

摘要:

目的 探讨在糖尿病大鼠中脐带间充质干细胞(UC-MSCs)能否通过调节自噬发挥肾脏保护作用.方法 选取36只8周龄雄性SD大鼠,通过单纯随机抽样分成正常对照组(control组,n=10)及高脂喂养组(n=26).采用高脂喂养联合小剂量链脲佐菌素腹腔注射诱导2型糖尿病大鼠模型,造模成功后继续高脂喂养8周,单纯随机抽样分为糖尿病组(T2DM组,n=10)和UC-MSCs治疗组(MSCs组,n=10).Control组一直予普通饲料喂养.MSCs组每2周给予1次UC-MSCs尾静脉输注(3×106/0.5 ml PBS),共治疗8周;同时T2DM组给予0.5 ml PBS溶液尾静脉输注,Control组不予处理.每2周监测随机血糖和体重.第四次治疗结束后1周所有大鼠行腹腔注射糖耐量实验,随后处死并留取肾脏组织,观察肾脏病理结构改变并检测自噬相关蛋白的表达.统计学分析采用两独立样本均数t检验、单因素方差分析和重复测量方差分析.结果 相较于T2DM组,MSCs组随机血糖下降[(23.9±3.7)比(28.7±2.1) mmol/L,t=3.568,P<0.01],糖耐量改善(t=3.07~6.64,P<0.01).病理结果显示,MSCs组肾小球硬化程度减轻,肾小球基底膜增厚缓解,肾脏间质纤维化程度改善(t=2.83~17.28,P<0.01).自噬相关蛋白检测结果显示:MSCs组LC3表达增加[(78.3±5.3)%比(33.4±8.3)%,t=10.22,P<0.01],P62表达减弱[(24.1±4.2)%比(93.3±4.9)%,t=24.13,P<0.01],提示MSCs输注以后大鼠肾脏自噬水平明显升高.结论 UC-MSCs治疗对糖尿病大鼠能发挥肾脏保护功能并且延缓糖尿病肾病的发生发展,其机制可能与UC-MSCs增强肾脏组织的自噬有关.

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abstracts:

Objective To investigate whether umbilical cord mesenchymal stem cells (UC-MSCs) can exert kidney protection in diabetes rats through regulating autophagy.Methods Type 2 diabetes rats model was induced by a high-fat diet combined with a low dosage of streptozotocin (STZ) intraperitoneal injection.After successful modeling,those rats were fed with a high-fat diet for another eight weeks,and the control group (10) was given a normal chow diet.The treatment group (MSCs group,10) was infused UC-MSCs (3 × 106 suspended in 0.5 ml PBS) via tail vein once every two weeks,for a total of eight weeks of treatment.The diabetes control group (T2DM group,10) was infused 0.5 ml PBS solution via tail vein and the control group did not receive any treatment at the same time.Random blood glucose and body weight were monitored every two weeks.One week after the fourth treatment,intraperitoneal glucose tolerance test (IPGTT) was performed in all rats,and later on they were sacrificed and their kidneys were collected for histopathological examinations and autophagy-related protein examinations.Data was measured using two independent sample mean t tests,one-way ANOVA and repeated measures analysis of variance.Results Compared to T2DM group,random blood glucose decreased [(23.9±3.7) vs (28.7±2.1) mmol/L,t=3.568,P<0.01],and glucose tolerance improved (t=3.07-6.64,P<0.01) in MSCs group.The pathological results showed that in the MSCs group the degree of glomerular sclerosis was reduced (P<0.01,t=17.28),the glomerular basement membrane thickening was relieved (P<0.01,t=12.00),and the degree of renal interstitial fibrosis was improved (P<0.01,t=2.83-17.28).Examinations of autophagy-related proteins showed that the MSCs group exhibited increased expression of LC3[(78.3±5.3)% vs(33.4±8.3)%,t=10.22,P<0.01] and reduced expression of P62[(24.1 ±4.2)% vs (93.3±4.9)%,t=24.13,P<0.01],which indicated that the level of autophagy in rat kidneys were significantly increased after MSCs infusion.Conclusion UC-MSCs therapy has the function of kidney protection and delay the development of diabetic nephropathy,which may be related to the enhancement of autophagy in renal tissues.

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作者: 龚正媛 [1] 李冰 [1] 张瀶曦 [1] 邹俊彦 [1] 于松岩 [1] 尹雅琪 [1] 薛婧 [1] 程愈 [1] 母义明 [1]
期刊: 《中华糖尿病杂志》2019年11卷6期 404-411页 ISTICPKUCSCD
栏目名称: 论著
DOI: 10.3760/cma.j.issn.1674-5809.2019.06.007
发布时间: 2019-08-13
基金项目:
国家自然科学基金面上项目(81870578)National Natural Science Foundation of China
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