摘要
背景糖尿病肾病是糖尿病最严重的并发症之一,长期高血糖会导致全身性的氧化应激和低度炎症状态。山柰酚是一种天然的黄酮类化合物,具有出众的抗炎和抗氧化的能力,可能会对糖尿病肾病的治疗有一定作用。目的研究山柰酚对糖尿病肾病小鼠肾损伤的治疗作用及其机制。方法18只6~8周龄FVB小鼠随机分为对照组、糖尿病模型组和山柰酚灌胃组,每组6只,并通过对模型组和山柰酚组腹腔注射链脲佐菌素构建1型糖尿病小鼠模型。模型建立后,治疗组灌胃山柰酚[70 mg/(kg·d)],对照组与模型组灌胃等量对照溶剂CMC-Na,16周后处死小鼠。病理染色观察小鼠肾病理损伤;qPCR和Western blot检测小鼠肾组织内炎症因子mRNA和氧化应激相关蛋白的表达;通过流式细胞术检测小鼠肾内巨噬细胞的数量和种类。体外建立高糖诱导肾小管上皮细胞(HK2)损伤模型,使用CCK-8试剂盒检测不同浓度山柰酚对HK2的活性影响,检测肾小管上皮细胞活性氧生成;qPCR和Western blot检测HK2内炎症因子mRNA和氧化应激相关蛋白以及炎症激活通路蛋白p38的表达。结果动物实验结果显示,与糖尿病模型组相比,经过灌胃山柰酚治疗后,肾病理损伤得到改善,肾小球系膜增生减少,足突融合减少;肾组织内白细胞介素(interleukin,IL)-1β、IL-6、肿瘤坏死因子α等炎症因子转录水平降低(P<0.01);NADPH氧化酶4表达降低(P<0.01);肾内炎性巨噬细胞数量减少,肾炎症微环境得到改善。细胞实验结果显示,山柰酚能够抑制高糖诱导下HK2的活性氧产生,改善氧化应激相关蛋白酶的表达(P<0.05);降低相关炎症因子mRNA的表达(P<0.01),且降低了p38的磷酸化(P<0.05)。结论山柰酚可能通过HO-1/p38通路减轻糖尿病肾小管上皮细胞炎症因子的分泌,增加相关氧化还原酶的表达,进而减轻糖尿病导致的肾损伤,保护肾功能。
Background Diabetic kidney disease(DKD)is one of the most serious vascular complications of diabetes mellitus and the main cause of end-stage renal disease.Long-term hyperglycemia leads to systemic oxidative stress and low-grade inflammation.Kaempferol,a natural flavonoid,has excellent anti-inflammatory and anti-oxidation abilities,and may play a role in the treatment of diabetic nephropathy.Objective To investigate the effects of kaempferol on renal injury and renal dysfunction in diabetic mice by reducing oxidative stress and inflammation.Methods Eighteen FVB mice aged 6-8 weeks were randomly divided into control group,diabetic model group and kaempferol gavage group.Type 1 diabetic mouse model was established by intraperitoneal injection of Streptozotocin(STZ).Blood glucose was measured every two weeks,and blood glucose over 16.6 mmol/L for two consecutive weeks was considered to be a successful diabetic model.After the model was established,the treatment group was given kaempferol(70mg/kg/d)by gavage,and the control group and the model group were given the same amount of CMC-Na by gavage.After 16 weeks for establishment of the model,the mice were sacrificed and the kidneys were obtained.The pathological injury of kidney was observed by pathological staining.qPCR and western blot were used to detect the mRNA expression of inflammatory factors and the expression of oxidative stress-related proteins in the kidney tissue of mice.The number and type of macrophages in the kidney were detected by flow cytometry.High glucose stimulated renal tubular epithelial cells(HK2)were used to establish a model of renal tubular injury in high glucose environment in vitro.The effect of different concentrations of kaempferol on the activity of HK2 was detected by CCK-8 kit.After high glucose stimulation and kaempferol treatment,ROS production in renal tubular epithelial cells was detected.The mRNA expression of inflammatory factors and the expression of oxidative stressrelated proteins in HK2 were detected by qPCR and western blot.Results The results of animal experiments showed that compared with the diabetic model group,after gavage of kaempferol,the renal pathological damage was significantly improved,the glomerular mesangial proliferation and foot process fusion were reduced.The transcription levels of inflammatory factors such as IL-1β,IL-6 and TNF-αin kidney tissue decreased significantly.The expression of NADPH oxidase 4 was significantly decreased.The number of inflammatory macrophages in the kidney was significantly reduced,and the renal inflammatory microenvironment was improved.The results of cell experiments showed that kaempferol could inhibit the production of reactive oxygen species(ROS)in HK2 induced by high glucose,and it significantly improved the expression of oxidative stress-related protease,while the mRNA expression of related inflammatory factors was significantly reduced.Conclusion Kaempferol can significantly reduce oxidative stress and inflammation in diabetic kidney tissue,reduce the secretion of inflammatory factors induced by high glucose,and increase the expression of related oxidoreductase in HK2,which may alleviate diabetic kidney injury through HO-1/p38 pathway.
作者
王超
魏翠婷
李润
佟琰
王雪
吴娇
欧阳清
陈香美
WANG Chao;WEI Cuiting;LI Run;TONG Yan;WANG Xue;WU Jiao;OUYANG Qing;CHEN Xiangmei(School of Traditional Chinese Medicine,Guangdong Pharmaceutical University,Guangzhou 510006,Guangdong Province,China;Department of Nephrology,the First Medical Center,Chinese PLA General Hospital,National Key Laboratory of Kidney Diseases,National Clinical Research Center for Kidney Diseases,Beijing Key Laboratory of Kidney Diseases Research,Beijing 100853,China;Department of Endocrinology,the First Medical Center,Chinese PLA General Hospital,Beijing 100853,China)
出处
《解放军医学院学报》
CAS
2024年第3期261-269,共9页
Academic Journal of Chinese PLA Medical School
基金
国家自然科学基金项目(32141005)。
