丹酚酸B抑制HMGB1/TLR4/NF-κB通路改善顺铂诱导的急性肾损伤

Effect of Salvianolic Acid B on Cisplatin Induced Acute Kidney Injury in Mice by Inhibiting HMGB1/TLR4/NF-κB Pathway

目的:探讨丹酚酸B对顺铂诱导小鼠急性肾损伤的影响,并探索其作用机制。方法:C57BL/6小鼠随机分为正常对照组、模型组及丹酚酸B低(50 mg/kg)、中(100 mg/kg)、高(200 mg/kg)剂量组。各组分别灌胃相应剂量丹酚酸B或生理盐水,1次/d,共5 d。第3天时除正常对照组腹腔注射生理盐水外,其余各组小鼠均以20 mg/kg腹腔注射顺铂建立小鼠急性肾损伤模型,48 h后处死小鼠。收集血清及肾组织,检测血清肌酐、尿素氮水平,HE染色观察肾组织病理学改变,TUNEL染色观察肾组织细胞凋亡情况,免疫组化检测肾组织HMGB1表达,Western Blot检测肾组织Bcl-2、Bax、HMGB1、TLR4、p-NF-κB p65蛋白表达。结果:与正常对照组比较,模型组血清肌酐、尿素氮水平显著升高,肾小管上皮细胞出现空泡样变性、小管管腔内多见管型,TUNEL染色可见较多阳性细胞,肾组织Bax/Bcl-2比值及HMGB1、TLR4、p-NF-κB p65蛋白表达显著升高(P<0.01)。与模型组比较,丹酚酸B中、高剂量组血清肌酐、尿素氮水平显著降低(P<0.05或P<0.01),肾小管管型减少,TUNEL染色阳性细胞较少,肾组织Bax/Bcl-2比值及HMGB1、TLR4、p-NF-κB p65蛋白表达显著降低(P<0.05或P<0.01)。结论:丹酚酸B可减轻顺铂诱导的小鼠急性肾损伤,其机制可能与抑制HMGB1/TLR4/NF-κB通路有关。

Objective:To investigate the effect of salvianolic acid B on cisplatin induced acute renal injury in mice and explore its mechanism.Methods:C57BL/6 mice were randomly divided into normal control group,model group and salvianolic acid B low(50 mg/kg),medium(100 mg/kg),high(200 mg/kg)dose groups.The mice in each group were given the corresponding dose of salvianolic acid B or normal saline once a day for a total of 5 days.On the third day,except for normal control group,mice in other groups were given 20 mg/kg cisplatin intraperitoneally injected to establish mice acute kidney injury model,and the mice were killed 48 hours later.The serum and kidney tissues were collected,serum creatinine and urea nitrogen levels were detected,renal histopathological changes were observed by HE staining,apoptosis of cells in kidney tissues was evaluated by TUNEL staining,HMGB1 expression in kidney tissue was detected by immunohistochemistry,and Bcl-2,Bax,HMGB1,TLR4,p-NF-κB p65 protein expression in kidney tissue were detected by Western Blot.Results:Compared with the normal control group,the levels of serum creatinine and urea nitrogen in the model group were significantly increased,vacuole-like degeneration of renal tubule epithelial cells,more tubule types were seen in the lumen of the tubule,more positive cells were detected by TUNEL staining,and the ratio of Bax/Bcl-2 and the expressions of HMGB1,TLR4 and p-NF-κB p65 proteins in kidney tissues were significantly increased(P<0.01).Compared with model group,the levels of serum creatinine and urea nitrogen in salvianolic acid B medium and high dose groups were significantly decreased(P<0.05 or P<0.01),the tubular type of renal tubules was decreased,there were fewer TUNEL positive cells,the ratio of Bax/Bcl-2 and the expressions of HMGB1,TLR4 and p-NF-κB p65 protein were significantly decreased(P<0.05 or P<0.01).Conclusion:Salvianolic acid B can alleviate cisplatin induced acute kidney injury in mice,and its mechanism may be related to inhibiting HMGB1/TLR4/NF-κB pathway.