基于α7nAChR-miRNA124-STAT3通路探讨肠缺血再灌注电针血清对腹腔巨噬细胞的影响  

Effect of Electroacupuncture Serum on Peritoneal Macrophages in Intestinal Ischemia-Reperfusion Injury based on α7nAChR-miRNA124-STAT3 Pathway

作　　者：耿艳霞[1] 裴颖皓 刘雨[2] 郭丁丁 李迎欣 郑晶晶 陈栋[1] GENG Yanxia;PEI Yinghao;LIU Yu;GUO Dingding;LI Yingxin;ZHENG Jingjing;CHEN Dong(The Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing Jiangsu 210029,China;Nanjing University of Chinese Medicine,Nanjing Jiangsu 210029,China)

机构地区：[1]南京中医药大学附属医院,江苏南京210029 [2]南京中医药大学,江苏南京210029

出　　处：《中医药导报》2024年第8期58-62,共5页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：江苏省自然科学基金项目(BK20210987);国家自然科学基金项目(82004456)。

摘　　要：目的:体外研究观察电针治疗肠缺血再灌注小鼠后,小鼠血清的效应物质干预腹腔巨噬细胞株RAW264.7的过程,探讨电针足三里改善肠缺血再灌注损伤(IIRI)的可能机制。方法:取BALB/c小鼠制备IIRI模型,随机分为正常血清组、模型血清组、电针血清组,电针血清组小鼠于足三里电针治疗30 min。各组小鼠制备动物血清后与RAW264.7细胞共培养,共设6组:对照组、正常血清组、模型血清组、电针血清组、电针血清+miRNA124抑制剂组、电针血清+α7nAChR拮抗剂组。实时荧光定量PCR法检测细胞α7nAChR和miRNA 124水平,Western blotting法检测细胞p-STAT3表达,ELISA法检测细胞上清IL-6浓度。结果:模型血清组p-STAT3及IL-6均较对照组和正常血清组显著增高;与模型血清组比较,电针血清组miRNA 124水平明显增高,而p-STAT3表达显著降低;与电针血清组比较,电针血清+α7nAChR拮抗剂组的α7nAChR水平显著下降,电针血清+miRNA 124抑制剂组和电针血清+α7nAChR拮抗剂组的miRNA 124水平显著下降,而p-STAT3和IL-6表达显著升高。结论:电针足三里可能通过促进巨噬细胞α7nAChR诱导miRNA 124,从而抑制STAT3通路改善IIRI。Objective:To explore the mechanism of electroacupuncture(EA)at Zusanli(ST 36)in the treatment of intestinal ischemia-reperfusion injury(IIRI)by observing the effect of mice EA serum on RAW264.7 cells in vitro.Methods:BALB/c mice were randomly divided into normal serum group,IIRI model serum group and EA serum group.Mice in the EA serum group were treated with EA at Zusanli(ST 36)for 30 min.Animal serum was prepared from each group of mice and co-cultured with RAW264.7 cells.There were 6 groups of cell models:control group,normal serum group,model serum group,EA serum group,EA serum+miRNA124 inhibitor group,and EA serum+α7nAChR antagonist group.The levels ofα7nAChR and miRNA 124 were detected by real-time fluorescence quantitative PCR.The expression of p-STAT3 was detected by Western blotting.The concentration of IL-6 in cell supernatant was detected by ELISA.Results:The levels of p-STAT3 and IL-6 in the model serum group were significantly higher than those in the control group and normal serum group.Compared with the model serum group,the level of miRNA 124 was significantly increased,and p-STAT3 was significantly decreased in the EA serum group.Compared with the EA serum group,the expression of miRNA 124 was decreased and the levels of p-STAT3 and IL-6 were increased in the miRNA 124 inhibitor group andα7nAChR antagonist group;while the level ofα7nAChR was significantly decreased in theα7nAChR antagonist group.Conclusion:EA at Zusanli(ST 36)may improve IIRI by promotingα7nAChR in macrophages to induce miRNA 124,thereby inhibiting STAT3 pathway.

关 键 词：电针血清 肠缺血再灌注 胆碱能抗炎通路 MIRNA 腹腔巨噬细胞 

分 类 号：R285.5[医药卫生—中药学]