氯化锂对脓毒症大鼠大脑皮质Caspase-3蛋白表达的影响  

Effect of lithium chloride on the expression of Caspase-3 protein in the cerebral cortex of rats with sepsis

作　　者：杨欣[1] 李熙鸿[2] 鲁利群[1] 黄莉[1] 李雅斐[2] YANG Xin LI Xihong LU Liqun HUANG Li L(Department of Pediatrics, Chengdu Medical School First Affiliated Hospi- tal, Sichuan Province, Chengdu 610500, China)

机构地区：[1]成都医学院第一附属医院儿科,610500 [2]四川大学华西第二医院儿童重症监护病房,610041

出　　处：《疑难病杂志》2016年第10期1061-1064,F0003,共5页Chinese Journal of Difficult and Complicated Cases

基　　金：教育部长江学者和创新团队发展计划(IRT09350);四川省科技厅支撑项目(12ZCO909)

摘　　要：目的观察氯化锂(LiCl)对脓毒症大鼠大脑皮质半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)蛋白表达的影响,探讨其对脓毒症脑损伤的保护作用机制。方法 2015年2—6月于四川大学华西第二医院进行实验,将Wistar大鼠60只按随机数字表法分为假手术组(S组,n=20)、模型组(CLP组,n=20)、氯化锂干预组(CLP+LiCl组,n=20),采用盲肠结扎穿孔术(CLP)制备脓毒症模型。CLP+LiCl组于CLP手术后立即给予腹腔注射LiCl溶液(50mg/kg),S组和CLP组给予等量生理盐水腹腔注射。CLP术后8 h、16 h、24 h每组取5只大鼠进行神经功能行为学评分的比较,而后采用断头法将大鼠处死取出脑组织皮质部分,采用Western blot法检测大鼠大脑皮质Caspase-3蛋白的表达;每组再于术后24 h各取5只大鼠断头取脑行免疫荧光法检测Caspase-3蛋白的表达。结果与S组比较,CLP组及CLP+LiCl组术后8、16、24 h大鼠神经功能行为学评分均有降低(F_(8h)=6.07,P=0.015;F_(16h)=14.10,P=0.001;F_(24h)=25.54,P=0.000),免疫荧光法检测大脑皮质Caspase-3蛋白的表达均增加,3组比较差异有统计学意义(分别为0.019 5±0.001 2、0.027 2±0.001 0、0.023 8±0.001 6,F=44.67,P<0.05);Western Blot法检测Caspase-3蛋白表达情况,3组差异有统计学意义(F¨=12.81,P=0.001;F_(16h)=33.01,P=0.000;F_(24h)=107.28,P=0.000);与CLP组同时间点比较,CLP+LiCl组神经功能行为学评分升高(除8 h外),而大脑皮质Caspase-3蛋白的表达下降,差异均有统计学意义(P<0.05)。结论氯化锂可能通过抑制大脑皮质神经元Caspase-3蛋白的表达而起到提高脓毒症大鼠神经行为学评分的作用。Objective To observe the effect of lithium chloride （LiC1） on the expression of Caspase-3 protein in the cerebral cortex of rats with sepsis, and to investigate the protective effect of lithium chloride on brain injury induced by sepsis. Methods Sixty male rats were randomly divided into sham operated group （S group, n = 20） , model group （CLP group, n = 20） ,and Lithium chloride treated group（CLP ＋ LiC1 group, n = 20）. Each group is divided into three subgroups by 8, 16, and 24 hours three time points. Models of rats with sepsis were established by cecal ligation and puncture （CLP）. In CLP ＋ LiCI group, LiCI solution（50 mg/kg）was injected into the rats peritoneal cavity after the operation of CLP immediately and e- qual saline solution was given to S group and CLP group by intraperitoneal injection. Five rats in each group were taken for neurologic behavior scores at 8 h, 16 h, and 24 h after CLP. Then these rats were broken the neck to death and the cortexes of brain tissue were taken for inspections. Western blotting was used to determine the expression of Caspase 3 at 8 h, 16 h, and 24 h after CLP. Immunohistochemistry staining also was used to investigate the expression of Caspase 3. Results Com- pared with S group, CLP group and CLP ＋ LiC1 group after 8, 16, 24 h rats neurological behavior scores were lower （ F 8 h = 6.07, P = 0. 015 ; F 16 h = 14.10, P = 0.001 ; F 24 h = 25.54, P = 0.000） to detect the expression of Caspase in cortex of im- munofluorescence 3 protein was increased, the difference between the 3 groups was statistically the significance of （0. 019 5±0. 001 2, 0.027 2 ± 0. 001 0, 0. 023 8 ± 0.001 6, F = 44.67, P 〈 0, 05） ; Western Blot 3 to detect Caspase protein expres- sion, there was significant difference between 3 groups （ F 8 h = 12. 81 , P = 0. 001 ; F 16 h = 33. 01, P = 0.000 ; F 24 h = 107. 28, P = 0. 000） ; CLP group compared with the same time point CLP ＋ LiC1 group, neurological behavior score increased （ ex- cept 8 h） , while t

关 键 词：氯化锂 脓毒症 脑损伤 半胱氨酸天冬氨酸蛋白酶-3 大鼠 

分 类 号：R459.7[医药卫生—急诊医学]