基于NF-κB信号通路研究当归芍药散对糖尿病肾病大鼠肾脏的保护作用及其机制

Protective Effect and Mechanism of Danggui Shaoyao Pulvis on Kidney of DiabeticNephropathy Rats Based on NF-κB Signaling Pathway

目的基于核因子(NF)-κB信号通路研究当归芍药散对糖尿病肾病(DN)大鼠肾脏的保护作用及其可能的通路机制。方法选择SPF级健康雄性SD大鼠,采用二联法(高脂高糖饮食+腹腔注射链脲佐菌素)诱导建立早期DN大鼠模型,随机分为模型对照组、当归芍药散组、厄贝沙坦组,同时设置空白组作为对照,各组给予相应的药物灌胃,分别干预4周。实验结束后记录大鼠体质量、随机血糖(GLU)、24 h饮水量及24 h尿量,分别对大鼠24 h尿蛋白、血肌酐值(Scr)、尿素氮(BUN)、肾脏肥大指数(KW/BW)等各项生理生化指标进行检测;对肾脏行苏木精-伊红(HE)、过碘酸希夫(PAS)染色,光镜下观察肾组织病理形态改变;酶联免疫吸附测定(ELISA)法检测血清中TNF-ɑ、IL-1β、IL-6炎症因子变化;免疫组化法检测肾脏组织中NF-κB蛋白的阳性表达。结果与空白对照组比较,模型对照组大鼠肾脏体积、KW/BW、GLU、24 h尿蛋白定量、24 h饮水量、24 h尿量、Scr、BUN、血清中TNF-ɑ、IL-1β、IL-6水平及肾脏组织NF-κB阳性表达升高(P<0.05),而体质量下降(P<0.05),且肾脏组织病理损害明显;与模型对照组比较,当归芍药散组大鼠肾脏体积、KW/BW、GLU、24 h尿蛋白定量、24 h饮水量、24 h尿量、Scr、血清中TNF-ɑ、IL-1β、IL-6水平及肾脏组织NF-κB阳性表达显著降低(P<0.05),而体质量增加(P<0.05),且肾脏组织病理损害有所缓解。结论当归芍药散可通过下调肾脏组织中炎症信号通路NF-κB蛋白的表达,减少TNF-ɑ、IL-1β、IL-6炎性细胞因子的释放,发挥对DN大鼠的肾脏保护作用,这可能是当归芍药散治疗DN的重要机制。

Objective This study aimed to investigate the protective effect of Danggui Shaoyao Pulvis on diabetic nephropathy(DN)in rats by examining the NF-κB signaling pathway and its potential mechanism.Methods Healthy male SD rats of SPF grade were selected and induced with an early DN rat model using a dual method(high-fat and high-sugar diet+intraperitoneal injection of Streptozotocin STZ);the rats were randomly divided into the model control group,Danggui Shaoyao Pulvis group,and irbesartan group,while a blank group was set as the control,Each group received the corresponding drugs via gavage for a 4-week intervention period.Various parameters such as body mass,random blood glucose(GLU),24 h water intake,24 h urine volume,24 h urine protein,blood creatinine(Scr),urea nitrogen(BUN),and kidney hypertrophy index(KW/BW)were recorded and measured.Kidney samples were stained with HE and PAS for observation under a light microscope.The serum levels of TNF-ɑ,IL-1β,and IL-6 inflammatory factors were detected using Elisa.The positive expression of NF-κB protein in renal tissues was determined by immunohistochemistry.Results Compared to the blank control group,the model control group exhibited increased kidney volume,KW/BW,GLU,24 h urinary protein quantity,24 h water intake,24 h urine volume,Scr,BUN,serum levels of TNF-ɑ,IL-1β,IL-6,and positive expression of NF-κB in kidney tissue(P<0.05),while body weight decreased(P<0.05).The kidney tissue also showed significant pathological injury.In contrast,the Danggui Shaoyao Pulvis group demonstrated significantly decreased kidney volume,KW/BW,GLU,24 h urinary protein quantity,24 h water intake,24 h urine volume,Scr,serum levels of TNF-ɑ,IL-1β,IL-6,and positive expression of NF-κB in kidney tissue(P<0.05),along with increased body mass(P<0.05).The pathological damage of kidney tissue was alleviated.Conclusions The findings of this study suggest that Danggui Shaoyao Pulvis can effectively reduce the release of TNF-ɑ,IL-1β,and IL-6 inflammatory cytokines in DN rats.This reduction is achieved by down-regulating the expression of NF-κB protein in the inflammatory signaling pathway of kidney tissue.These results indicate that the modulation of NF-κB protein expression may be a crucial mechanism by which Danggui Shaoyao Pulvis exerts its therapeutic effects in treating DN.