美洲大蠊研粉干预脊髓半横断大鼠运动功能和自噬蛋白Beclin-1的表达  

Effects of Periplaneta americana powder on motor function and autophagic protein Beclin-1 in rats undergoing spinal cord hemisection

作　　者：周邦瑜 李杰 阮玉山 耿福能 李绍波 Zhou Bangyu;Li Jie;Ruan Yushang;Geng Funeng;Li Shaobo(Clinical Medicine Collage,Dali University,Dali 671000,Yunnan Province,China;Department of Spine Surgery,the First Affiliated Hospital of Dali University,Dali 671000,Yunnan Province,China;Sichuan Key Laboratory of Medicinal American Cockroach,Xichang 615000,Sichuan Province,China;Department of Surgery,Clinical Medicine College,Dali University,Dali 671000,Yunnan Province,China)

机构地区：[1]大理大学临床医学院,云南省大理市671000 [2]大理大学第一附属医院脊柱外科,云南省大理市671000 [3]药用美洲大蠊四川省重点实验室,四川省西昌市615000 [4]大理大学临床医学院外科教研室,云南省大理市671000

出　　处：《中国组织工程研究》2024年第8期1223-1228,共6页Chinese Journal of Tissue Engineering Research

基　　金：云南省科技厅高校联合项目-面上项目(2019FH001-015),项目负责人:李绍波;云南省教育厅科学研究基金项目(2022Y865),项目负责人:周邦瑜;大理大学第一附属医院杰出中青年人才基金项目(DFYJC-202114),项目负责人:阮玉山。

摘　　要：背景:脊髓损伤有氧化应激、炎症、凋亡、自噬等机制参与其中,激活自噬可改善脊髓损伤后的神经运动功能,对脊髓起保护作用。目的:探讨美洲大蠊研粉对脊髓半横断损伤大鼠后肢运动功能和损伤区域自噬蛋白Beclin-1的影响。方法:取30只6-8周龄SD大鼠,采用随机数字表法分为3组,每组10只:假手术组仅打开椎板,暴露脊髓后缝合;生理盐水组与美洲大蠊组均建立左侧脊髓半横断损伤模型,术后第1天开始分别灌胃给予生理盐水与美洲大蠊研粉,连续给药14 d。术后6 h及1,3,7,14 d行BBB评分,观察后肢运动功能的情况;给药14 d后处死大鼠,采用免疫组化染色、Western Blot检测、免疫荧光染色检测脊髓半横断损伤区域自噬蛋白Beclin-1的表达情况。结果与结论:①随着术后时间的延长,生理盐水组、美洲大蠊组大鼠BBB评分逐渐升高;与假手术组比较,生理盐水组、美洲大蠊组术后6 h及1,3,7,14 d的BBB评分均降低(P<0.05);美洲大蠊组术后7,14 d的BBB评分高于生理盐水组(P<0.05);②免疫组化染色显示,假手术组Beclin-1呈弱阳性表达,主要位于胞浆;生理盐水组Beclin-1主要位于胞浆,可见部分核膜表达;美洲大蠊组Beclin-1主要位于胞浆,可见部分核膜表达;生理盐水组、美洲大蠊组Beclin-1阳性细胞比例多于假手术组(P<0.05),美洲大蠊组Beclin-1阳性细胞比例多于生理盐水组(P<0.05);③Western Blot检测与免疫荧光染色显示,生理盐水组、美洲大蠊组Beclin-1蛋白表达高于假手术组(P<0.05),美洲大蠊组Beclin-1蛋白表达高于生理盐水组(P<0.05);④结果表明,美洲大蠊研粉能够改善脊髓半横断损伤大鼠的后肢运动功能,机制可能是美洲大蠊研粉中的多糖类物质增加了自噬蛋白Beclin-1的表达。BACKGROUND:Spinal cord injury involves mechanisms such as oxidative stress,inflammation,apoptosis and autophagy.Activation of autophagy can improve neuromotor function after spinal cord injury and play a protective role in the spinal cord.OBJECTIVE:To investigate the effects of Periplaneta americana powder on hindlimb motor function and the autophagy protein Beclin-1 in the injured site of rats after spinal cord hemisection.METHODS:Thirty Sprague-Dawley rats,6-8 weeks of age,were randomly divided into three groups(n=10 per group).In the sham-operated group,the lamina was just opened to exposure the spinal cord followed by suturing.Normal saline group and Periplaneta americana powder group both underwent left hemisection of the spinal cord to prepare animal models of spinal cord hemisection.The normal saline group was continuously gavaged with normal saline for 14 days,and the Periplaneta americana powder group was continuously gavaged with Periplaneta americana powder for 14 days.The Basso Beattie Bresnahan scale score was performed at the 6th hour,1st day,3rd day,7th day and 14th day after operation to observe the hindlimb motor function.After 14 days of administration,the rats were sacrificed and sampled.Immunohistochemistry,western blot and immunofluorescence were used to detect the expression of Beclin-1 in the injured site of the spinal cord after hemisection.RESULTS AND CONCLUSION:After operation,the Basso Beattie Bresnahan scale scores were gradually increased in the normal saline group and Periplaneta americana powder group.Compared with the sham-operated group,the Basso Beattie Bresnahan scale scores were significantly reduced in the normal saline group and Periplaneta americana powder group at the 6th hour,1st day,3rd day,7th day and 14th day after operation(P<0.05).The Basso Beattie Bresnahan scale scores in the Periplaneta americana powder group were significantly higher than those in the normal saline group at the 7th and 14th days after operation(P<0.05).Immunohistochemical staining showed that Becl

关 键 词：脊髓损伤 脊髓半横断损伤 美洲大蠊研粉 自噬 BECLIN-1 运动功能 

分 类 号：R459.9[医药卫生—治疗学] R313[医药卫生—临床医学] R683.2