摘要
目的评价乌司他丁对周围神经损伤小鼠脊髓神经元凋亡的影响。方法SPF级健康雄性C57BL/6J小鼠225只,6~8周龄,体重18~22g,采用随机数字表法分为3组(n=75):假手术组(Sham组)、周围神经损伤组(PNI组)和乌司他丁组(UTI组)。PNI组和UTI组建立单侧坐骨神经损伤模型,造模成功后UTI组腹腔注射乌司他丁10000U/kg,Sham组和PNI组给予等容量生理盐水,1次/d,连续3d。于给药后1、3、7、14和28d(T0-4)时处死小鼠取脊髓L4-6节段,光镜下观察病理学结果,采用TUNEL法检测神经元凋亡情况,计算神经元凋亡指数(AI)。于T0-4时采用Western blot法检测脊髓caspase一12、Bcl-2、Bax和活化型caspase-3的表达,计算Bcl-2/Bax比值,采用RT—PCR法检测脊髓caspase-12 mRNA表达。结果与Sham组比较,PNI组和UTI组AI升高,脊髓Bcl-2表达下调,caspase。12、活化型caspase-3和Bax表达上调,Bcl-2/Bax比值降低(P<0.05);与PNI组比较,UTI组AI降低,脊髓Bcl-2表达上调,caspase-12、活化型caspase-3和Bax表达下调,Bcl-2/Bax比值升高(P<0.05)。UTI组脊髓病理学损伤较PNI组减轻。结论乌司他丁可抑制周围神经损伤小鼠脊髓神经元凋亡。
Objective To evaluate the effect of ulinastatin on neuronal apoptosis in spinal cord after peripheral nerve injury in mice.Methods A total of 225healthy male SPF C57BL/6J-mice,aged 6-8 weeks,weighing 18-22g,were divided into 3groups (n =75 each)using a random number table method: sham operation group (group Sham),peripheral nerve injury group (group PNI)and ulinastatin group (group UTI).The model of unilateral sciatic nerve transection was established in PNI and UTI groups.Ulinastatin 10000U/kg was injected intraperitoneally once a day for 3consecutive days in group UTI,while the equal volume of normal saline was given instead in Sham and PNI groups.The mice were sacrificed at 1,3,7,14 and 28 days after administration (T0-4),and the L4-6 segments of the spinal cord were removed for examination of the pathological changes (under the light microscope)and for determination of neuronal apoptosis (by TUNEL)and expression of caspase-12,Bcl-2,Bax and activated caspase-3(by Western blot)and expression of caspasc-12 mRNA (by real-time polymerase chain reaction).Apoptosis index (AI)and Bcl-2/Bax ratio were calculated.Results Compared with group Sham,AI was significantly increased,the expression of Bcl-2 was down-regulated,the expression of caspase-12,activated caspase-3 and Bax was up-regulated,and Bcl-2/Bax ratio was decreased in PNI and UTI groups (P<0.05).Compared with group PNI,AI was significantly decreased,the expression of Bcl-2 was up-regulated,the expression of caspase-12,activated caspase-3 and Bax was down-regulated,and Bcl-2/Bax ratio was increased in group UTI (P<0.05).The pathological changes of the spinal cord were significantly attenuated in group UTI than in group PNI.Conclusion Ulinastatin can inhibit neuronal apoptosis in the spinal cord after peripheral nerve injury in mice.
作者
窦忠慈
张丽霞
张卫
Dou Zhongci;Zhang Lixia;Zhang Wei(Department of Anesthesiology,First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2018年第9期1099-1102,共4页
Chinese Journal of Anesthesiology
