摘要
目的观察自噬基因相关蛋白在大鼠脾脏撞击伤后12 h内脾组织内的表达情况。方法选用清洁级健康SD大鼠20只,随机分为正常对照组(A组)、创伤后2 h组(B组)、创伤后8 h组(C组)、创伤后12 h组(D组),每组5只。B、C、D组利用生物撞击机建立脾脏撞击伤模型,A组只麻醉不撞击,麻醉后立即处死取出脾脏,B、C、D组分别于撞击后相应时间取出脾脏,评估脾脏出现损伤的程度。创伤脾组织标本进行HE染色,显微镜下观察受损脾组织的特征。大体标本观察造模是否成功。采用免疫组织化学法及Western blot法检测各组脾组织酵母自噬相关基因Atg6的同源物(Beclin1)、酵母自噬相关基因Atg8的同源物微管相关蛋白1轻链蛋白3(LC3)蛋白的表达。结果 (1)脾撞击伤模型均造模成功。同一撞击力度下不同受伤时间模型组大鼠脾脏裂伤长度和深度基本一致(P均> 0. 05)。(2) HE染色显示模型组脾组织符合脾破裂征象。(3)免疫组织化学法及Western blot法检测显示,模型大鼠随创伤后时间推移(2 h组→8 h组→12 h组),其脾组织Beclin1、LC3Ⅱ蛋白相对表达量递升(P均<0. 05),且模型大鼠各时间组均高于对照组(P均<0. 05)。结论脾脏撞击伤可导致脾组织自噬水平的上升;脾脏撞击伤后12 h内脾组织自噬相关蛋白水平随时间的推移,表达逐渐升高。
Objective To observe the expressions of autophagy gene related-proteins in spleen tissues within 12 hours after splenic impact injury in rats. Methods Twenty clean grade healthy SD rats were randomly divided into four groups( n = 5 each) : control group( group A),2 hours after trauma group( group B),8 hours after trauma group( group C) and12 hours after trauma group( group D). The splenic impact injury models were established by bio-impactor in rats of group B,C and D. The rats in group A were anesthetized only but not impacted and immediately executed after anesthesia,then the spleens were taken out. The rat spleens were taken out at the corresponding time after impact,and the extent of spleen injury was assessed in group B,C and D. Hematoxylin eosin( HE) staining of traumatic spleen was performed to observe the characteristics of injured spleen tissues under microscope. Gross specimens were observed to see if the molding was successful. Immunohistochemical method and Western blot method were used to detect the expressions of Beclin1( a homologue of yeast autophagy related gene Atg6) and microtubule-associated protein 1 light chain3( LC3,a homologue of yeast autophagy related gene Atg8 proteins). Results The models of splenic impact injury were established successfully.Gross specimens showed that plenic laceration length and depth under the same impact force and at different injured time in each modeling group were basically same( all P > 0. 05). HE staining showed that splenic tissues conformed to signs of splenic rupture. Immunohistochemical method and Western blot method showed that the relative expression levels of Beclin1 and LC3Ⅱ proteins in spleen increased progressively with posttraumatic time going( 2 h→8 h→12 h) in model group( all P < 0. 05),and they in each time subgroup of model group were significantly higher than those in control group( all P <0. 05). Conclusion Spleen impact injury could lead to the rise of autophagy level in spleen tissues,and the autophagy related proteins levels within 12 hours after splenic impact injury increases progressively with time going.
作者
欧阳骏骏
牛玉林
0UYANG Jun-jun;NIU Yu-lin(Department of Pediatric Surgery,Anqing Hospital Affiliated to Anhui Medical University,Anqing,Anhui 246000,China)
出处
《中国临床研究》
CAS
2019年第1期18-22,共5页
Chinese Journal of Clinical Research
基金
贵州省科技合作计划项目(黔科合LH字[2015(7399号)]~~
