芍药苷抗急性坏死性胰腺炎相关性肾损伤的作用与机制研究

Protective effect of paeoniflorin against renal injury induced by acute necrotizing pancreatitis and its mechanism

背景与目的:芍药苷是传统中药抗炎的重要药物,其具有广谱抗炎作用,而在急性坏死性胰腺炎(ANP)相关肾损伤中其是否发挥抗炎作用从而保护肾功能尚不明确,因此,本研究探讨芍药苷对ANP相关肾损伤中的影响及作用机制。方法:将SD大鼠随机分为假手术组、ANP模型组(ANP组)、ANP模型+芍药苷处理组(芍药苷组),ANP模型采用胆胰管逆行注射5%的牛磺胆酸钠诱导,芍药苷在造模后通过股静脉注射。首先观察不同剂量芍药苷(50、100、150 mg/kg)对ANP大鼠血清淀粉酶(AMY)、脂肪酶(LIPA)及肝功能指标的影响,分析芍药苷的最适剂量。随后采用最适剂量的芍药苷进行干预,检测造模后不同时间点(3、6、12 h)各组大鼠胰腺与肾脏组织病理变化、血清炎性指标水平、AMY与LIPA及尿素氮(BUN)与肌酐(Cr)水平。在各组造模后12 h肾脏组织中,用免疫组化检测NF-κB与caspase-3的表达、TUNEL法检测细胞凋亡、Western blot检测p38与磷酸化p38(p-p38)的表达。结果:量效关系分析结果,100 mg/kg为芍药苷的最适剂量。后续实验结果显示,与假手术组比较,ANP组与芍药苷组造模后均出现不同程度的ANP与肾损伤病理变化(病理评分增加)、炎性指标(TNF-α、IL-1β、IL-6)水平升高、血清酶学指标(AMY、LIPA)以及肾功能指标(BUN、Cr)升高,且均随着时间延长而加剧,但芍药苷组的各项指标在各时间点均明显低于ANP组(均P<0.05)。在造模后12 h肾脏组织中,ANP组与芍药苷组NF-κB与caspase-3表达、凋亡细胞数、p-p38/p38比例均明显升高,但芍药苷组的升高程度明显低于ANP组(均P<0.05)。结论:芍药苷有明显抗ANP相关肾损伤作用,其作用机制一方面在于抑制ANP本身的进展,另一方面可能通过降低p38通路活性抑制NF-κB相关炎症因子所致的级联瀑布反应从而减轻ANP相关肾损伤。

Background and Aims:Paeoniflorin is an important traditional Chinese medicine with broad-spectrum anti-inflammatory properties.However,its role in protecting kidney function in acute necrotizing pancreatitis(ANP)-related renal injury remains unclear.Therefore,this study was conducted to investigate the effects of paeoniflorin on ANP-related renal injury and its action mechanism.Methods:SD rats were randomly divided into a sham surgery group,an ANP model group(ANP group),and an ANP model plus paeoniflorin treatment group(paeoniflorin group).ANP was induced by retrograde injection of 5%sodium taurocholate into the biliopancreatic duct,and paeoniflorin was injected via the femoral vein after modeling.Initially,different doses of paeoniflorin(50,100,150 mg/kg)were tested to determine the optimal dosage based on their effects on serum amylase(AMY),lipase(LIPA),and liver function indicators.Subsequently,the optimal dose of paeoniflorin was used for intervention,and at different time points after modeling(3,6,12 h),the groups were evaluated for pancreatic and renal tissue histopathological changes,serum inflammatory markers,AMY,LIPA,urea nitrogen(BUN),and creatinine(Cr)levels.In renal tissues 12 h after modeling of all groups,immunohistochemistry was employed to examine the expression of NF-κB and caspase-3,TUNEL assay was used to detect cell apoptosis,and Western blot was performed to analyze the expression of p38 and phosphorylated p38(p-p38).Results:The dose-response analysis revealed that 100 mg/kg was the optimal dosage of paeoniflorin.Subsequent experiments showed that compared to the sham surgery group,both the ANP group and the paeoniflorin group exhibited varying degrees of ANP and renal injury histopathological changes(increased histopathological scores),elevated inflammatory markers(TNF-α,IL-1β,IL-6),increased serum enzyme indicators(AMY,LIPA)and renal function indicators(BUN,Cr)after modeling,which worsened over time.However,the paeoniflorin group showed significantly lower levels of all these variables at each time point than the ANP group(all P<0.05).In renal tissues 12 h after modeling,the expression of NF-κB and caspase-3,the number of apoptotic cells,and the p-p38/p38 ratio were significantly increased in both the ANP group and the paeoniflorin group;however,the extents of increases in the paeoniflorin group was significantly lower than those in the ANP group(all P<0.05).Conclusion:Paeoniflorin exerts significant protective effects against ANP-related renal injury.Its mechanism of action involves inhibiting the progression of ANP on the one hand,and on the other hand,it may reduce ANP-related renal damage by suppressing the cascade reaction caused by NF-κB-related inflammatory factors through the reduction of p38 pathway activity.