苦杏仁苷对糖尿病肾病大鼠肾小管上皮细胞损伤的影响  

Effects of amygdalin on renal tubular epithelial cell injury in diabetes nephropathy rats

作　　者：闫萌萌 徐林丽 刘岩岩[3] 张艳[4] YAN Meng-meng;XU Lin-li;LIU Yan-yan;ZHANG Yan(Internal Medicine Teaching and Research Office,Cangzhou Medical College,Cangzhou 061000,Hebei Province,China;Surgery Teaching and Research Office,Cangzhou Medical College,Cangzhou 061000,Hebei Province,China;Pediatric Teaching and Research Office,Cangzhou Medical College,Cangzhou 061000,Hebei Province,China;Science and Technology Experimental Center,Cangzhou Medical College,Cangzhou 061000,Hebei Province,China)

机构地区：[1]沧州医学高等专科学校内科教研室,河北沧州061000 [2]沧州医学高等专科学校外科教研室,河北沧州061000 [3]沧州医学高等专科学校儿科教研室,河北沧州061000 [4]沧州医学高等专科学校科技实验中心,河北沧州061000

出　　处：《中国临床药理学杂志》2024年第7期1019-1023,共5页The Chinese Journal of Clinical Pharmacology

基　　金：河北省沧州市重点研发指导基金资助项目(192106021)。

摘　　要：目的研究苦杏仁苷对糖尿病肾病(DKD)大鼠肾小管上皮细胞损伤及腺苷酸活化蛋白激酶/哺乳动物西罗莫司靶蛋白(AMPK/mTOR)通路的影响。方法将SD大鼠分为空白组、模型组、苦杏仁苷低剂量组(3mg·kg^(-1))、苦杏仁苷高剂量组(10mg·kg^(-1))、CompoundC组(2.5mg·kg^(-1)AMPK抑制药CompoundC)、苦杏仁苷高剂量+CompoundC组(10mg·kg^(-1)苦杏仁苷+2.5mg·kg^(-1)/CompoundC),每组10只。除空白组外,其余各组大鼠用连续喂养高脂高糖饲料+腹腔注射60mg·kg^(-1)1%链脲佐菌素(STZ)方法进行DKD造模,造模成功后第2天给予相应药物处理,每天1次,持续给予8周,空白组、模型组均给予等体积0.9%NaCl。用二喹啉甲酸(BCA)法检测大鼠24h尿蛋白含量,用血糖仪检测大鼠空腹血糖,用全自动生化分析仪检测大鼠血肌酸酐和血尿素氮水平,用原位末端标记法(TUNEL)染色检测大鼠肾组织细胞凋亡情况,用酶联免疫吸附测定(ELISA)法检测大鼠血清中肿瘤坏死因子-α(TNF-α)、白细胞介素1β(IL-1β)水平,用蛋白质印迹(WesternBlot)法测定大鼠肾组织AMPK/mTOR通路、自噬及凋亡蛋白的表达水平。结果空白组、模型组、苦杏仁苷低剂量组、苦杏仁苷高剂量组、Compound C组和苦杏仁苷高剂量+CompoundC组的肾小管损伤评分分别为(0.43±0.05)、(3.26±0.24)、(1.92±0.14)、(0.73±0.06)、(3.81±0.29)和(1.78±0.12)分,细胞凋亡率分别为(5.23±0.41)%、(26.35±2.17)%、(16.76±1.42)%、(6.13±0.53)%(35.68±3.08)%和(15.92±1.15)%,磷酸化mTOR(p-mTOR)/mTOR分别为0.31±0.03、0.80±0.08、0.60±0.06、0.38±0.04,0.92±0.08和0.69±0.07,磷酸化AMPK(p-AMPK)/AMPK分别为0.91±0.08、0.39±0.04、0.56±0.06、0.88±0.07、0.23±0.02和0.52±0.05。模型组的上述指标与空白组、苦杏仁苷低剂量组、苦杏仁苷高剂量组相比,在统计学上差异均有统计学意义(均P<0.05);苦杏仁苷高剂量组的上述指标与苦杏仁苷高剂量+CompoundC组相比,在统计学上�Objective To study the effect of amygdalin on renal tubular epithelial cell injury and adenosine monophosphate activated protein kinase/mammalian target of rapamycin(AMPK/mTOR)pathway in diabetic kidney disease(DKD)rats.Methods SD rats were grouped into blank group,model group,low dose amygdalin group(3 mg·kg^(-1)),high dose amygdalin group(10 mg·kg^(-1)),Compound C group(AMPK inhibitor Compound C 2.5 mg·kg^(-1)),and high dose amygdalin+Compound C group(10 mg·kg^(-1)amygdalin+2.5 mg·kg^(-1)Compound C),with 10 rats in each group.Except for blank group,the rats in other groups were fed with high-fat and high-glucose diet continuously+intraperitoneal injection of 60 mg·kg^(-1)1%streptozotocin(STZ)to construct the rat model of DKD.After the success of the model,the rats were given corresponding drug treatment the second day,once a day for 8 weeks,and the blank group and the model group were given equal volume physiological saline.Bicinchoninic acid(BCA)method was applied to detect 24-hour urine protein content in rats;glucose meter was applied to detect fasting blood glucose of rats;automatic biochemical analyzer was applied to detect the serum creatinine level and blood urea nitrogen level of rats;terminal-deoxynucleoitidyl transferase mediated nick end labeling(TUNEL)staining was applied to detect apoptosis in kidney tissue cells;enzyme linked immunosorbent assay(ELISA)was applied to detect the levels of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)in serum;and Western Blot was used to measure the AMPK/mTOR pathway,autophagy,and apoptotic protein expression levels in kidney tissue.Results The renal tubule injury scores in blank group,model group,low dose amygdalin group,high dose amygdalin group,0.73±0.06,3.81±0.29 and 1.78±0.12,respectively;the apoptosis rates were(5.23±0.41)%,(26.35±2.17)%,(16.76±1.42)%,(6.13±0.53)%,(35.68±3.08)%and(15.92±1.15)%,respectively;phosphorylated mTOR(p-mTOR)/mTOR were 0.31±0.03,0.80±0.08,0.60±0.06,0.38±0.04,0.92±0.08 and 0.69±0.07,respectively;phospho

关 键 词：苦杏仁苷 腺苷酸活化蛋白激酶/哺乳动物西罗莫司靶蛋白 糖尿病肾病 肾小管上皮细胞 

分 类 号：R28[医药卫生—中药学]