半胱氨酰天冬氨酸特异性蛋白酶-3和Beclin1在腹部创伤大鼠组织的表达及其意义  

Expression levels and significance of cysteinyl aspartate-specific protease-3 and Beclin1 in abdominal trauma rats

作　　者：阿尔帕提·买买提[1] 贾德政 郭飞[1] Alpati·Maimaiti;Jia Dezheng;Guo Fei(Department of Emergency Trauma Surgery,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830001,China)

机构地区：[1]新疆医科大学第一附属医院急诊创伤外科,乌鲁木齐830001

出　　处：《中华实验外科杂志》2024年第4期724-727,共4页Chinese Journal of Experimental Surgery

基　　金：新疆维吾尔自治区自然科学基金(2019D01C304)。

摘　　要：目的探讨半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3和Beclin1在腹部创伤大鼠组织中的表达及意义。方法20只SD大鼠建立腹部创伤大鼠模型,按照数字表法分为对照组和腹部创伤组,采用荧光定量聚合酶链反应(PCR)和蛋白质免疫印迹分析两组大鼠肠道组织Caspase-3和Beclin1,采用原位缺口末端标记法(TUNEL)染色分析两组大鼠肠道组织细胞凋亡水平;采用免疫荧光分析两组大鼠肠道组织LC3点的数量;采用免疫组织化学分析两组大鼠肠道组织裂解的Caspase-3(cleaved-Caspase-3)蛋白表达水平。酶联免疫吸附试验(ELISA)检测两组大鼠肠道组织炎性因子水平。组间计量数据比较采用t检验。结果对照组大鼠肠道组织Caspase-3和Beclin1 mRNA表达水平(0.96±0.11、1.08±0.12)明显低于腹部创伤组大鼠(1.82±0.12、1.77±0.18),差异有统计学意义(t=16.910、10.120,P<0.05)。对照组大鼠肠道组织炎性因子肿瘤坏死因子-α(TNF-α)表达水平[(18.82±3.56)pg/ml]明显低于腹部创伤组大鼠[(71.98±11.80)pg/ml],差异有统计学意义(t=13.640,P<0.05)。对照组大鼠肠道组织Caspase-3和Beclin1蛋白相对表达水平(0.60±0.08、0.89±0.08)明显低于腹部创伤组大鼠(1.34±0.18、1.56±0.16),差异有统计学意义(t=12.170、12.230,P<0.05)。对照组大鼠肠道组织cleaved Caspase-3蛋白相对表达水平(97.66±11.25)明显低于腹部创伤组大鼠(191.33±24.47),差异有统计学意义(t=10.990,P<0.05)。对照组大鼠肠道组织TUNEL染色阳性率[(2.83±1.10)%]明显低于腹部创伤组大鼠[(50.59±10.15)%],差异有统计学意义(t=14.790,P<0.05)。对照组大鼠肠道组织LC3点数量[(45.20±14.91)个]明显低于腹部创伤组大鼠[(82.81±13.17)个],差异有统计学意义(t=5.977,P<0.05)。蛋白免疫印迹结果显示,对照组大鼠肠道组织自噬底物p62蛋白表达水平(1.26±0.11)明显高于腹部创伤组大鼠(0.62±0.11),差异有统计学意义(t=12.940,P<0.05)。结论腹部创伤后Objective To investigate the expression levels and significance of cysteinyl aspartate-specific protease(Caspase)-3 and Beclin1 in abdominal trauma rats.Methods A total of 20 SD rats were divided into control group and abdominal trauma group by a digital table method.Caspase-3 and Beclin1 in the intestinal tissues of the two groups were analyzed by fluorescence quantitative polymerase chain reaction(PCR)and Western blotting,and the apoptosis level of the intestinal tissues of the two groups was analyzed by terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)staining.The number of LC3 points in intestinal tissues of the two groups was analyzed by immunofluorescence.The expression of cleaved Caspase-3 protein in the intestinal tissues of the two groups was analyzed by immunohistochemistry.The levels of intestinal inflammatory factors were detected by enzyme linked immunosorbent assay(ELISA).T test was used to compare measurement data between groups.Results The mRNA expression levels of Caspase-3 and Beclin1 in the intestinal tissue of rats in the control group(0.96±0.11,1.08±0.12)were significantly lower than those in the abdominal trauma group(1.82±0.12,1.77±0.18;t=16.910,10.120,P<0.05).The expression level of inflammatory factor tumor necrosis factor-α(TNF-α)in intestinal tissue of rats in control group[(18.82±3.56)pg/ml]was significantly lower than that of rats in abdominal trauma group[(71.98±11.80)pg/ml,t=13.640,P<0.05].The relative expression levels of Caspase-3 and Beclin1 in intestinal tissue of rats in the control group(0.60±0.08,0.89±0.08)were significantly lower than those in the abdominal trauma group(1.34±0.18,1.56±0.16;t=12.170,12.230,P<0.05).The relative expression level of intestinal cleaved Caspase-3 protein in the control group(97.66±11.25)was significantly lower than that in the abdominal trauma group(191.33±24.47,t=10.990,P<0.05).The positive rate of TUNEL staining in intestinal tissue of rats in control group[(2.83±1.10)%]was significantly lower than that in ab

关 键 词：半胱氨酰天冬氨酸特异性蛋白酶-3 BECLIN1 腹部创伤 凋亡 自噬 

分 类 号：R656[医药卫生—外科学]