温和灸联合间充质干细胞移植对大鼠肛门括约肌损伤的修复作用及其机制  

Effect and mechanism of mild moxibustion combined with mesenchymal stem cell transplantation on repair of anal sphincter injury in rats

作　　者：金文琪 郭修田[1] 朱影 李鹏 李小嘉 JIN Wenqi;GUO Xiutian;ZHU Ying;LI Peng;LI Xiaojia(Anorectal Department,Shanghai Hospital of Traditional Chinese Medicine,Shanghai 200071,China)

机构地区：[1]上海市中医医院肛肠科,上海200071

出　　处：《山西医科大学学报》2023年第2期215-221,共7页Journal of Shanxi Medical University

基　　金：上海市卫生和计划生育委员会基金项目(20174Y0018);国家自然青年科学基金项目(81804110)。

摘　　要：目的探讨温和灸联合间充质干细胞移植对大鼠肛门括约肌损伤修复、α-SMA水平及Wnt信号通路的影响。方法6~8周龄雄性SD大鼠被随机分为假手术组、模型组、温和灸组、间充质干细胞(BMSCs)组、温和灸+BMSCs组,每组10只。除假手术组外,其余大鼠均建立肛门括约肌损伤模型,建模成功后,温和灸组采用温和灸治疗,BMSCs组大鼠括约肌损伤处及缺损两侧断端边缘注射107个BMSCs,温和灸+BMSCs组在注射107个BMSCs后采用温和灸治疗,模型组注射同体积生理盐水。采用多道信息采集处理系统检测括约肌功能(检测指标包括收缩压、静息压、肌电振幅及肌电频率);HE染色观察肛门括约肌病理形态;免疫组化检测α-SMA阳性表达;免疫印迹检测Wnt3a、Wnt5a、β-catenin蛋白水平。比较5组大鼠治疗后7,14,21 d肛门括约肌病理形态及各观察指标水平。结果与假手术组相比,模型组治疗后7,14,21 d收缩压、静息压、肌电振幅及肌电频率均明显降低(P<0.05)。与模型组相比,温和灸组治疗后7,14,21 d的收缩压、静息压、肌电振幅及肌电频率升高(P<0.05);温和灸组和BMSCs组治疗后7,14,21 d的收缩压、静息压、肌电振幅及肌电频率比较差异无统计学意义(P>0.05);与BMSCs组相比,温和灸+BMSCs组治疗后7,14,21 d的收缩压、静息压、肌电振幅及肌电频率均升高(P<0.05)。与假手术组相比,模型组α-SMA阳性细胞数降低(P<0.05);与模型组相比,温和灸组α-SMA阳性细胞数升高(P<0.05),温和灸+BMSCs组α-SMA阳性细胞数差异无统计学意义(P>0.05);与BMSCs组相比,温和灸+BMSCs组在α-SMA阳性细胞数升高(P<0.05)。与假手术组相比,模型组Wnt3a、Wnt5a、β-catenin明显降低(P<0.05);与模型组相比,温和灸组Wnt3a、Wnt5a、β-catenin升高(P<0.05);温和灸组和BMSCs组间Wnt3a、Wnt5a、β-catenin差异无统计学意义(P>0.05);与BMSCs组相比,温和灸+BMSCs组Wnt3a、Wnt5a、β-catenin均升高(P<0.05)。�Objective To investigate the effect of mild moxibustion combined with mesenchymal stem cell transplantation on the repair of anal sphincter injury,α-SMA level and Wnt signaling pathway in rats.Methods Male SD rats aged 6-8 weeks were randomly divided into sham operation group,model group,mild moxibustion group,mesenchymal stem cells(BMSCs)group and mild moxibustion+BMSCs group,with 10 rats in each group.After successful modeling,the rats in mild moxibustion group were treated with mild moxibustion,the rats in BMSCs group were injected with 107 BMSCs at the anal sphincter injury site and the edge of the broken end on both sides of the defect,and the rats in mild moxibustion+BMSCs group were injected with mild moxibustion after 107 BMSCs injection.The rats in model group were injected with the same volume of normal saline.Multi-channel information acquisition and processing system was used to detect the sphincter function(systolic blood pressure,resting blood pressure,electromyography amplitude and electromyography frequency).HE staining was used to observe the pathological morphology of anal sphincter.The positive expression ofα-SMA was detected by immunohistochemistry.Western blot was used to detect the protein levels of Wnt3a,Wnt5a andβ-catenin.The pathological morphology of anal sphincter and the levels of the above indexes were compared among the five groups at day 7,14 and 21 after treatment.Results Compared with sham operation group,systolic blood pressure,resting blood pressure,electromyography amplitude and electromyography frequency in model group were significantly decreased at day 7,14 and 21 after treatment(P<0.05).Compared with model group,systolic blood pressure,resting blood pressure,electromyography amplitude and electromyography frequency were increased in mild moxibustion group at day 7,14 and 21 after treatment(P<0.05).There was no statistically significant difference in systolic blood pressure,resting blood pressure,electromyography amplitude and electromyography frequency between mild moxib

关 键 词：肛门括约肌损伤 温和灸 间充质干细胞移植 肌肉修复 WNT信号通路 Α-平滑肌肌动蛋白 大鼠 

分 类 号：R657.12[医药卫生—外科学]