壳聚糖纳米颗粒负载落新妇苷通过调控MAPK14/HSP27影响高糖诱导的肾小管上皮细胞铁死亡  

Chitosan nanoparticles loaded withastilbin affect high glucose-induced ferroptosis of renal tubular epithelial cells by regulating MAPK14/HSP27

作　　者：陈毅君[1] 李湘[2] 胡剑卓[1] Chen Yijun;Li Xiang;Hu Jianzhuo(Dept of Nephrology and Endocrinology,2Dept of Emergency,Second Affiliated Hospital of Hunan University of Traditional Chinese Medicine,Changsha 410005)

机构地区：[1]湖南中医药大学第二附属医院肾病内分泌科,长沙410005 [2]湖南中医药大学第二附属医院急诊科,长沙410005

出　　处：《安徽医科大学学报》2024年第9期1610-1620,共11页Acta Universitatis Medicinalis Anhui

基　　金：湖南省自然科学基金(编号:2022JJ70026)。

摘　　要：目的探讨壳聚糖纳米颗粒(CS NPs)负载落新妇苷(落新妇苷-CS-NPs)通过调控丝裂原活化蛋白激酶14(MAPK14)/热休克蛋白27(HSP27)对高糖(HG)诱导的肾小管上皮细胞铁死亡的影响。方法将HK-2细胞分组如下:正常糖(NG)组、HG组、HG+落新妇苷-CS-NPs(0、5、10、20 mg/L)组,以及HG条件下的pcDNA3.1-MAPK14组、pcDNA3.1-MAPK14+落新妇苷-CS-NP组、si-HSP27组、pcDNA3.1-MAPK14+si-HSP27组、pcDNA3.1-MAPK14+si-HSP27+落新妇苷-CS-NPs组。CCK-8法检测细胞活力,TUNEL法检测细胞凋亡。同时通过试剂盒测定铁离子水平、乳酸脱氢酶(LDH)活性、谷胱甘肽(GSH)水平、活性氧(ROS)水平。构建糖尿病肾脏病(DKD)大鼠模型,并使用落新妇苷-CS-NPs干预,探究其对体内DKD的影响。结果与NG组相比,HG组HK-2细胞活力降低、凋亡增加,铁离子水平、LDH活性、ROS水平升高,而GSH水平明显降低(P<0.05)。经过落新妇苷-CS-NPs处理组明显逆转了HG对HK-2细胞生物学行为及铁死亡相关指标的影响。此外,相比于pcDNA3.1组,pcDNA3.1-MAPK14组铁死亡增加,而敲减HSP27或者联合落新妇苷-CS-NPs干预则改善了这一情况。体内实验结果表明,落新妇苷-CS-NPs能够改善DKD大鼠肾损伤、抑制铁离子水平以及MAPK14/HSP27的表达。结论落新妇苷-CS-NPs可能通过抑制MAPK14与HSP27表达,改善HG诱导的肾小管上皮细胞铁死亡。Objective To explore the effect of astilbin(astilbin-CS-NPs)-loaded chitosan nanoparticles(CS-NPs)on high glucose(HG)-induced ferroptosis of renal tubular epithelial cells via regulating mitogen-activated protein kinase 14(MAPK14)/heat shock protein 27(HSP27).Methods HK-2 cells were divided into the following groups:normal glucose(NG)group,HG group,HG+astilbin-CS-NPs(0,5,10,20 mg/L)group,and under HG condition,pcDNA3.1-MAPK14 group,pcDNA3.1-MAPK14+astilbin-CS-NP group,si-HSP27 group,pcDNA3.1-MAPK14+si-HSP27 group,and pcDNA3.1-MAPK14+si-HSP27+astilbin-CS-NPs group.Cell viability was detected using CCK-8 assay,cell apoptosis was detected via Tunel assay.Meanwhile,iron ion levels,lactate dehydrogenase(LDH)activity,glutathione(GSH)levels,and reactive oxygen species(ROS)levels were measured using assay kits.Rat model of diabetic kidney disease(DKD)was constructed and intervened with astilbin-CS-NPs to explore the effects of astilbin-CS-NPs on DKD in vivo.Results Compared with the NG group,the HK-2 cell viability in the HG group was significantly reduced,apoptosis increased,iron ion levels,LDH activity,and ROS levels were significantly elevated,while GSH levels significantly decreased(all P<0.05).Treatment with astilbin-CS-NPs significantly reversed the effects of HG on the biological behavior of HK-2 cells and ferroptosis-related indicators.Additionally,compared to the pcDNA3.1 group,the pcDNA3.1-MAPK14 group showed increased ferroptosis,which was improved by knocking down HSP27 or co-intervention with astilbin-CS-NPs.In vivo experimental results showed that astilbin-CS-NPs could improve DKD rat kidney injury,inhibit iron ion levels and the expression of MAPK14/HSP27.Conclusion Astilbin-CS-NPs may improve HG-induced ferroptosis of renal tubular epithelial cells via inhibition of MAPK14 and HSP27 expression.

关 键 词：落新妇苷 壳聚糖纳米颗粒 丝裂原活化蛋白激酶14 热休克蛋白27 铁死亡 糖尿病肾病 

分 类 号：R692.9[医药卫生—泌尿科学]