小檗胺对糖尿病肾病大鼠肾损伤和炎性细胞因子产生的影响

Effects of berbamine on renal injury and inflammatory cytokines production in diabetic nephropathy rats

目的研究小檗胺(berbamine,BBM)对糖尿病肾病(diabetic nephropathy,DN)大鼠的保护作用及其机制。方法将大鼠分为对照组组、模型组、DN+BBM组(50、100、200 mg/kg),采用高脂喂食结合化学诱导法建立DN大鼠模型。检测24 h尿蛋白含量,检测血清肌酸酐(SCr)和尿素氮(BUN);苏木精-伊红(HE)和TUNEL染色观察肾脏病理情况;Western blot检测凋亡、炎症通路相关蛋白表达;ELISA检测血清中炎症细胞因子表达。建立LPS+DN大鼠模型,检测指标同上。结果与模型组相比,DN+BBM组大鼠尿蛋白、血清肌酐和尿素氮含量、肾组织病理损伤、凋亡率、cl-caspase-3、cl-caspase-9和Bax蛋白表达、炎症细胞因子IL-1β、IL-6和TNF-α含量、炎症通路蛋白p-P65、MIP-2表达均降低;Bcl-2表达增加。此外,BBM显著减弱LPS对DN大鼠肾脏功能、凋亡率及相关蛋白表达、血清炎性细胞因子及炎症通路蛋白表达的调节作用。结论小檗胺能有效缓解糖尿病肾病大鼠肾损伤和炎症反应,其机制与抑制NF-κB p65活化有关。

Diabetic nephropathy(DN)is one of the most important complications of diabetes mellitus and has become the main cause of end-stage renal disease.This study was designed to investigate the protective effect of berbamine(BBM)on diabetic nephropathy(DN)rats,and its mechanism.Rats were randomly divided into control group,DN model group,and DN+BBM(50,100 and 200 mg/kg)groups.DN rat model was established by high-fat diet combined with chemical induction.24 hours urine was collected to detect urinary protein content,while blood was taken to detect serum creatinine(SCr)and urea nitrogen(BUN);Hematoxylin-eosin(HE)and TUNEL staining were used to observe renal pathology;Western blot was used to detect the expression of apoptosis-and inflammatory pathway-related proteins.ELISA was used to detect the expression of inflammatory cytokines in serum.Furthermore,LPS+DN rat model was established,and all indexes mentioned above were detected and analyzed.Data showed that as compared with DN group,rats in DN+BBM group demonstrated lower levels of urinary protein,serum creatinine and urea nitrogen,alleviated pathological damage of kidney tissue,lower levels of apoptotic rate,cl-caspase-3,cl-caspase-9 and Bax proteins expression,and lower levels of IL-1β,IL-6,TNF-α,and inflammatory pathway proteins,but higher level of Bcl-2.In additional,BBM significantly attenuated the regulation of LPS on renal function,apoptotic rate and related protein expression,serum inflammatory cytokines and inflammatory pathway protein expression in DN rats.In conclusion,Berbamine can effectively alleviate renal injury and inflammation in diabetic nephropathy rats,and its mechanism is related to the inhibition of NF-κB p65 activation.