摘要
目的探究环磷酰胺调控过氧化物酶体增殖物激活受体-γ(peroxisome proliferators-activated receptor-γ,PPAR-γ)介导足细胞自噬减轻糖尿病肾病(diabetic nephropathy,DN)小鼠肾损伤的作用及机制。方法60只小鼠分为对照组、模型组、环磷酰胺组、环磷酰胺+PPAR-γ抑制剂组,每组15只小鼠。除对照组外,其余各组小鼠给予高脂饲料喂养8周,一次性腹腔注射35 mg/kg链脲佐菌素构建DN小鼠模型;成模后,环磷酰胺组小鼠腹腔注射30 mg/kg环磷酰胺,环磷酰胺+PPAR-γ抑制剂组小鼠腹腔注射30 mg/kg环磷酰胺和1 mg/kg PPAR-γ抑制剂GW9662,连续10周。测量小鼠体质量与肾脏质量;全自动生化仪检测小鼠24 h尿蛋白(24 h-urinary proteins,24 h UP)与血清中血糖(blood glucose,BG)、甘油三酯(triglyceride,TRIG)、胆固醇(cholesterol,CHOL)、血肌酐(serum creatinine,SCr)和尿素氮(blood urea nitrogen,BUN)的水平;HE染色和Masson染色检测肾组织病理形态学变化与纤维化程度;透射电子显微镜下观察肾组织足细胞自噬情况;ROS荧光探针-二氢乙啶(DHE)法检测肾组织ROS水平;Western blot检测肾组织PPAR-γ蛋白、自噬相关蛋白及AMPK/mTOR信号通路相关蛋白的表达水平。结果与对照组比较,模型组小鼠体质量和肾脏质量均显著减少(P<0.05);24 h UP水平显著升高,血清BG、TRIG、CHOL、SCr和BUN水平也显著升高(P<0.05);肾小管萎缩、坏死与空泡样,肾小球基底膜增厚,肾组织明显纤维化;足细胞自噬体与凋亡小体减少,Beclin-1蛋白表达与LC3Ⅱ/LC3Ⅰ比值显著下调,P62蛋白表达显著上调(P<0.05);肾组织PPAR-γ蛋白表达下调(P<0.05);p-AMPK/AMPK比值显著下降,p-mTOR/mTOR比值则显著升高(P<0.05)。与模型组比较,环磷酰胺组小鼠体质量和肾脏质量有所增加(P<0.05);24 h UP水平与血清BG、TRIG、CHOL、SCr及BUN水平均显著下降(P<0.05);肾组织病理形态学变化得到明显改善,纤维化程度减小;足细胞自噬体与凋亡小体�
Objective To explore the effect of cyclophosphamide regulating peroxisome proliferators-activated receptors-γ(PPAR-γ)-mediated autophagy on kidney injury of diabetic nephropathy(DN)mice and its mechanism.Methods Sixty mice were divided into control group,model group,cyclophosphamide group,cyclophosphamide+PPAR-γinhibitor group,with 15 mice in each group.Except for control group,the mice in the other groups were fed with high-fat diet for 8 weeks,and intraperitoneally injected with 35 mg/kg streptozotocin to construct DN mouse models.After modeling,mice in cyclophosphamide group were injected intraperitoneally with 30 mg/kg cyclophosphamide,and mice in cyclophosphamide+PPAR-γinhibitor group were injected intraperitoneally with 30 mg/kg cyclophosphamide and 1 mg/kg PPAR-γinhibitor GW9662 for 10 consecutive weeks.The body mass and the kidney mass of the mice were measured.Levels of 24 h-urinary proteins(24 h UP)and serum blood glucose(BG),triglyceride(TRIG),cholesterol(CHOL),serum creatinine(SCr)and blood urea nitrogen(BUN)were detected by an automatic biochemical analyzer.The pathological changes and fibrosis degree of kidney tissues were detected by HE staining and Masson staining.The autophagy of kidney tissue podocytes was observed under transmission electron microscope.The level of ROS in kidney tissue was detected by the ROS fluorescent probe-dihydroethiidine(DHE)method.The expression levels of PPAR-γprotein,autophagy-related proteins and AMPK/mTOR signaling pathway-related proteins in kidney tissues were determined by Western blot.Results Compared with control group,the body weight and the kidney weight were significantly reduced in model group(P<0.05),while 24 h UP level and serum BG,TRIG,CHOL,SCr,BUN levels were significantly increased(P<0.05).Compared with control group,the renal tubules were found atrophy,necrosis and vacuole-like phenomena in model group,the glomerular basement membrane was thickened,and the kidney tissue was obviously fibrotic.Compared with control group,the podocyte autophagosome
作者
李燕
李安琪
余晓洋
张文静
吕佳
王志刚
张亚莉
陈蕾
LI Yan;LI Anqi;YU Xiaoyang;ZHANG Wenjing;L Jia;WANG Zhigang;ZHANG Yali;CHEN Lei(Department of Kidney Medicine,First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,China;Department of Respiration,First Affiliated Hospital of Xi’an Jiaotong University;Department of Blood Purification,First Affiliated Hospital of Xi’an Jiaotong University)
出处
《山西医科大学学报》
CAS
2021年第8期1021-1028,共8页
Journal of Shanxi Medical University
基金
国家自然科学基金资助项目(81900675)。
关键词
糖尿病肾病
环磷酰胺
过氧化物酶体增殖物激活受体-Γ
自噬
活性氧
diabetic nephropathy
cyclophosphamide
peroxisome proliferator-activated receptor-γ
autophagy
reactive oxygen species
