腹腔注射木犀草素调控sirt1/ FOXO1通路对慢性坐骨神经结扎大鼠痛觉敏化的影响  

Effect of luteolin intraperitoneal injection on hyperalgesia in rats with chronic sciatic nerve constriction injury by regulating sirt1/FOXO1 pathway

作　　者：华汤锋 金红芳[2] 吕晨[3] 郭小文[3] HUA Tangfeng;JIN Hongfang;LYU Chen;GUO Xiaowen(Department of Anesthesiology,Xiaoshan District Hospital of Traditional Chinese Medicine,Hangzhou 311201,China)

机构地区：[1]杭州市萧山区中医院麻醉科,311201 [2]杭州市萧山区第一人民医院消化内科 [3]浙江省中医院麻醉科

出　　处：《浙江医学》2021年第14期1489-1493,1512,共6页Zhejiang Medical Journal

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+EX5270.5 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。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 1d 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 d14 after surgery,and the expression of sirt1,FOXO1 and acetylated FOXO1 proteins were determined by Western blot.Results Behavioral test results showed that MWT and TWL in each group were similar on 1d 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 difference in TWL compared with S group on d7 and d14 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 FOXO1 protein in spinal dorsal horn were up-regulated in all groups on 7 and 14 days after C

关 键 词：木犀草素 sirt1/FOXO1通路 神经病理性疼痛 痛觉敏化 

分 类 号：R285.5[医药卫生—中药学]