基于内质网应激诱导的细胞凋亡探究黄蜀葵花总黄酮减轻糖尿病肾小管病的作用和机制  

Effects and mechanisms of total flavones of Abelmoschus manihot in attenuating diabetic tubulopathy by targeting endoplasmic reticulum stress-induced cell apoptosis

作　　者：万冰莹 曹东维 万毅刚[4] 陈岱[2] 吴薇[4] 房其军[4] 刘思逸 涂玥 王玉[1] WAN Zi-yue WAN Bing-ying;CAO Dong-wei;WAN Yi-gang;CHEN Dai;WU Wei;FANG Qi-jun;LIU Si-yi;TU Yue;WANG Yu;WAN Zi-yue(Department of Traditional Chinese Medicine,Nanjing Drum Tower Hospital Clinical College of Nanjing University of Chinese Medicine,Nanjing 210008,China;Changzhou Hospital of Traditional Chinese Medicine Affiliated to Nanjing University of Chinese Medicine,Changzhou 213000,China;Department of Nephrology,Shanghai Municipal Hospital of Traditional Chinese Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200071,China;Department of Traditional Chinese Medicine,Nanjing Drum Tower Hospital,the Affiliated Hospital of Nanjing University Medical School,Nanjing 210008,China;Department of Traditional Chinese Medicine Health Preservation,Acupuncture,Moxibustion and Massage College,Health Preservation and Rehabilitation College,Nanjing University of Chinese Medicine,Nanjing 210023,China;Graduate School of Social Sciences,Faculty of Social Sciences,Hitotsubashi University,Tokyo 186-8601,Japan)

机构地区：[1]南京中医药大学鼓楼临床医学院中医科,江苏南京210008 [2]南京中医药大学常州附属医院,江苏常州213000 [3]上海中医药大学附属市中医医院肾病科,上海200071 [4]南京大学医学院附属鼓楼医院中医科,江苏南京210008 [5]南京中医药大学针灸推拿学院·养生康复学院,江苏南京210023 [6]Graduate School of Social Sciences,Faculty of Social Sciences,Hitotsubashi University,Tokyo 186-8601

出　　处：《中国中药杂志》2023年第10期2657-2666,共10页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金面上项目(81573903,82174472);江苏省自然科学基金面上项目(BK20211298);江苏省中医药科技发展计划项目(MS2022071,MS2021037);江苏省研究生科研与实践创新计划项目(KYCX22_2065);南京中医药大学自然科学基金项目(XZR2021087);南京市名中医专家工作室建设项目;南京鼓楼医院中西医协同特色技术发展项目(CZXM2022098)。

摘　　要：糖尿病肾脏疾病(diabetic kidney disease, DKD)患者的肾小管损伤可能是与肾小球和微血管病变同时出现的,并且在其肾损伤进展中发挥着重要的作用,目前被称之为“糖尿病肾小管病(diabetic tubulopathy, DT)”。为了探究传统治肾中药黄蜀葵花的提取物——黄蜀葵花总黄酮(total flavones of Abelmoschus manihot,TFA)在体内减轻DT的多靶点治疗作用和药理机制,笔者将全部大鼠随机分为4组,即正常对照组(正常组)、DT模型组(模型组)、DT模型+TFA组(TFA组)以及DT模型+罗格列酮(rosiglitazone, ROS)组(ROS组)。通过综合措施建立基于DKD的大鼠DT模型。在DT模型鼠成模后,每天用双蒸水、TFA混悬液、ROS混悬液分别给4组大鼠连续灌胃。在给药6周后,处死全部大鼠,分别收集其尿液、血液以及肾组织等标本而进行各项指标的检测,并观察TFA和ROS对DT模型鼠血液、尿液生化指标,肾小管损伤指标,肾小管上皮细胞凋亡和内质网应激(endoplasmic reticulum stress, ERS)指标以及肾组织蛋白激酶样内质网激酶(protein kinase-like endoplasmic reticulum kinase, PERK)-真核翻译起始因子2α(eukaryotic translation initiation factor 2α,eIF2α)-激活转录因子4(activating transcription factor 4,ATF4)-同源蛋白(C/EBP homologous protein C/EBP,CHOP)信号通路活性的影响。结果表明,DT模型鼠出现了肾小管上皮细胞肥大,肾小管增生、闭塞以及肾间质细胞外基质和胶原沉积;同时,还出现了明显的肾小管损伤标志分子表达程度和蛋白表达水平的变化;此外,出现了肾小管性尿蛋白的异常升高。经TFA和ROS干预,模型鼠尿蛋白、肾小管损伤特征、肾小管上皮细胞凋亡和ERS特征以及肾组织PERK-eIF2α-ATF4-CHOP信号通路活性均得到不同程度的改善;其中,对于肾小管/间质病理性改变,TFA的作用优于ROS。简言之,借助DT模型鼠,该研究阐明,TFA在体内能通过抑制肾小管ERS诱导的细胞凋亡而多靶点�Renal tubular injury in patients with diabetic kidney disease(DKD)may be accompanied by glomerular and microvascular diseases.It plays a critical role in the progression of renal damage in DKD,and is now known as diabetic tubulopathy(DT).To explore the multi-targeted therapeutic effects and pharmacological mechanisms in vivo of total flavones of Abelmoschus manihot(TFA),an extract from traditional Chinese medicine for treating kidney disease,in attenuating DT,the authors randomly divided all rats into four groups:a normal control group(normal group),a DT model group(model group),a DT model+TFA-treated group(TFA group)and a DT model+rosiglitazone(ROS)-treated group(ROS group).The DT rat model was established based on the DKD rat model by means of integrated measures.After successful modeling,the rats in the four groups were continuously given double-distilled water,TFA suspension,and ROS suspension,respectively by gavage every day.After 6 weeks of treatment,all rats were sacrificed,and the samples of their urine,blood,and kidneys were collected.The effects of TFA and ROS on various indicators related to urine and blood biochemistry,renal tubular injury,renal tubular epithelial cell apoptosis and endoplasmic reticulum stress(ERS),as well as the activation of the protein kinase R-like endoplasmic reticulum kinase(PERK)-eukaryotic translation initiation factor 2α(eIF2α)-activating transcription factor 4(ATF4)-C/EBP homologous protein(CHOP)signaling pathway in the kidney of the DT model rats were investigated.The results indicated that hypertrophy of renal tubular epithelial cells,renal tubular hyperplasia and occlusion,as well as interstitial extracellular matrix and collagen deposition occurred in the DT model rats.Moreover,significant changes were found in the expression degree and the protein expression level of renal tubular injury markers.In addition,there was an abnormal increase in tubular urine proteins.After TFA or ROS treatment,urine protein,the characteristics of renal tubular injury,renal tubular epithe

关 键 词：糖尿病肾小管病 黄蜀葵花总黄酮 内质网应激 细胞凋亡 PERK-eIF2α-ATF4-CHOP信号通路 

分 类 号：R285.5[医药卫生—中药学]