【摘要】 目的探讨腹腔注射木犀草素调控sirt1/FOXO1通路对慢性坐骨神经结扎(CCI)大鼠痛觉敏化的影响。方法将72只SD雄性大鼠按随机数字表法分为假手术组(S组)、慢性坐骨神经结扎组(CCI组)、CCI联合木犀草素腹腔给药组(Lut组)、CCI联合木犀草素和沉默信息调节因子1(sirt1)抑制剂EX725腹腔给药组(Lut+EX组),每组各18只。S组仅暴露坐骨神经并游离连接组织,不做CCI,其余3组大鼠建立CCI模型。模型建立后,S组和CCI组腹腔注射0.9%氯化钠注射液2.5 ml/kg,Lut组腹腔注射木犀草素2.0 ml/kg+含1%二甲基亚砜(DMSO)的0.9%氯化钠注射液0.5 ml/kg,Lut+EX组腹腔注射木犀草素2.0 ml/kg+EX527 0.5 ml/kg,均连续给药7 d。分别在术前1天和CCI模型建立后第1、3、7、14天测定各组大鼠的机械性缩足反射阈值(MWT)和热缩足潜伏期(TWL)。术前1天和CCI后第7、14天提取大鼠脊髓背角组织,测定sirt1、叉头转录因子1(FOXO1)和乙酰化FOXO1蛋白表达。结果各组大鼠CCI前1天MWT和TWL均相近(均P>0.05);CCI后第1、3、7、14天,CCI组、Lut组、Lut+EX组MWT和TWL均明显低于S组(均P<0.05),但Lut组第7、14天TWL与S组比较差异无统计学意义(P>0.05);CCI后第1、3、7、14天,Lut组MWT和TWL均明显高于CCI组(均P<0.05),而Lut+EX组与CCI组比较差异均无统计学意义(均P>0.05)。与S组比较,各组大鼠CCI后第7、14天脊髓背角sirt1蛋白水平均下降(均P<0.05),Lut组和Lut+EX组sirt1蛋白水平较CCI组明显增高(均P<0.05)。与S组相比,各组大鼠在CCI后第7、14天脊髓背角FOXO1和乙酰化FOXO1蛋白水平均上调(均P<0.05);与CCI组、Lut+EX组比较,Lut组在CCI后第7天FOXO1蛋白水平升高,第14天FOXO1蛋白水平降低,乙酰化FOXO1水平较前两组降低更明显(均P<0.05)。结论木犀草素可上调CCI大鼠脊髓背角sirt1蛋白水平,降低FOXO1乙酰化,通过调节sirt1/FOXO1信号通路,降低神经病理性疼痛导致的痛觉敏化。更多还原

【Abstract】 Objective To investigate the effect of luteolin(Lut) intraperitoneal injection on hyperalgesia in rats with chronic sciatic nerve constriction injury and its relation with sirt1/FOXO1 pathway.Methods Seventy two male SD rats were randomly divided into four groups with 18 animals in each group.The sciatic nerve was ligated in chronic constriction injury(CCI)group,Lut group and Lut+EX group,and the sham operation was performed in S group.In Lut group and Lut+EX group intraperitoneal injection of 100 mg/kg luteolin or 100 mg/kg luteolin and 5 mg/kg EX527 were given continuously for 7 days after the CCI model established,for CCI and S groups intraperitoneal injection of 2.5 ml/kg NS was given instead.Mechanical withdrawal threshold(MWT) and thermal foot withdrawal latency(TWL) were measured by Von Frey filament,on 1 d before CCI surgery and d1,d3,d7 and d14 after surgery,respectively.The spinal dorsal horn of rats was extracted 1 d before surgery and d7 and d 14 after surgery,and the expression ofsirt1,FOXO 1 and acetylated FOXO 1 proteins were determined by Western blot.Results Behavioral test results showed that MWT and TWL in each group were similar on 1 d before CCI(P>0.05).On 1,3,7 and 14 days after CCI,MWT and TWL in CCI group,Lut group and Lut+EX group were lower than those in S group(P<0.05).However,in Lut group,there was no significant difere nc e in TWL compared with S group on d7 and a 14 after CCI(P>0.05).On 1,3,7 and 14 days after CCI,MWT and TWL in Lut group were significantly higher than those in CCI group(P<0.05),while there were no significance differences between Lut+EX group and CCI group(P<0.05).The sirt1 protein levels in spinal dorsal horn were decreased in all groups after CCI compared with those in S group(P<0.05).On 7 and 14 days after CCI,the protein level of sirt1 in Lut group and Lut+EX group was significantly higher than that in CCI group(all P<0.05).Compared with group S,the levels of FOXO1 and acetylated FOXO 1 protein in spinal dorsal horn were up-regulated in all groups on 7 and 14 days after CCI(P<0.05).Compared with CCI group and Lut+EX group,the FOXO1 protein level in Lut group was significantly decreased at 7 and 14 days after CCI,especially the acetylated FOXO1 level,which was significantly lower than that in the other two groups(P<0.05).Conclusion Luteolin can up-regulate the level of sirt1 in the spinal dorsal horn of CCI rats,reduce FOXO1 acetylation,and inhibit the hyperalgesia ofneuropathic pain by regulating the sirt1/FOXO1 signaling pathway.更多还原