标题：蛙血清小分子肽对大鼠脑缺血再灌注细胞凋亡的影响

      作者：陈博 1，谢琳 2
    (广东医学院东莞校区生理科学实验室 1、基础医学院 2，广东 东莞 523808)

      卷次： 2016年27卷7期

      【摘要】 目的 探讨蛙血清小分子肽对大鼠脑缺血再灌注损伤后细胞凋亡及凋亡相关蛋白的作用，分析其可能
的作用机制。方法 将雄性SD大鼠100只，按随机数字表法分为假手术组、模型组和蛙血清小分子肽高(90 mg/kg)、
中(30 mg/kg)、低(10 mg/kg)剂量组，每组20只。采用改良线栓法栓塞大鼠大脑中动脉建立大鼠缺血再灌注损伤模
型，缺血2 h再灌注24 h，进行神经行为评分，后处死；采用TUNEL法检测细胞凋亡；采用免疫组化法检测B细胞淋巴
瘤/白血病-2 (Bcl-2)、Bcl相关x蛋白(Bax)、细胞色素 c (Cytc)、半胱氨酰天冬氨酸特异性蛋白酶3 (Caspase-3)的表达
情况。结果 在神经行为评分上，假手术组为(0.00±0.00)分，模型组为(2.38±0.78)分，小分子肽高剂量组为(1.47±0.41)
分，中剂量组为(1.68±0.52)分，低剂量组为(2.01±0.66)分，小分子肽高、中剂量组与模型组比较差异均有统计学意义
(P<0.05)；在凋亡细胞计数上，假手术组为(3.37±1.10)个，模型组为(42.80±3.54)个，小分子肽高剂量组为(28.00±2.28)
个，中剂量组为(32.40±3.26)个，低剂量组为(41.40±1.20)个，小分子肽高、中剂量组与模型组比较差异均有统计学意
义(P<0.05)；在凋亡相关蛋白 Bax、Cytc、Caspase-3和 Bcl-2表达上(个/高倍视野)，假手术组分别为(3.01±1.12)、
(2.58±1.74)、(2.34±1.37)、(65.42±3.65)，模型组分别为(70.67±3.06)、(58.31±5.04)、(68.04±5.85)、(31.26±2.81)，小分
子肽高剂量组分别为 (40.56±4.52)、(33.65±3.44)、(41.56±4.52)、(55.64±5.49)，中剂量组分别为 (47.29±5.04)、
(38.09±4.24)、(47.29±5.04)、(48.33±4.26)，低剂量组分别为(53.20±4.70)、(44.53±4.39)、(53.20±4.70)、(40.35±3.17)，小
分子肽高、中、低剂量组均能抑制Bax、Cytc、Caspase-3蛋白表达，增加Bcl-2蛋白表达，与模型组比较差异有统计学
意义(P<0.05)，以高剂量组效果最优，且存在量效依赖关系。结论 蛙血清小分子肽是通过增强Bcl-2蛋白表达和
抑制Bax、Cytc、Caspase-3蛋白表达来减少神经元细胞凋亡数目，改善神经功能症状，保护脑缺血再灌注损伤。

      【关键词】 大鼠；蛙血清；小分子肽；脑缺血再灌注；细胞凋亡；作用机制

      【中图分类号】 R-332 【文献标识码】 A 【文章编号】 1003—6350（2016）07—1036—03


Effects of frog serum small molecular peptide on apoptosis in rats with cerebral ischemia reperfusion injury.
CHEN Bo 1, XIE Lin 2.
Laboratory of Physiological Science 1, Basic Medical College 2, Dongguan Campus, Guangdong
Medical University, Dongguan 523808, Guangdong, CHINA

【Abstract】 Objective To investigate the effect of small molecular peptide of frog serum on apoptosis and apopto-
sis related proteins in rats with cerebral ischemia-reperfusion injury, and to analyze the possible mechanism. Methods A
total of 100 male SD rats were randomly divided into five groups according to the random number table: sham operation
group, model group and frog serum small peptide with high dose group (90 mg/kg, high dose group), middle dose group
(30 mg/kg), low dose group (10 mg/kg), with 20 rats in each group. Using artery suture embolization in the rat brain to
establish the rat model of ischemia reperfusion injury, the rats were scored for neurological behavior after ischemia 2 h
reperfusion 24 h and then sacrificed. Apoptosis was detected by TUNEL method. Immunohistochemical method were ap-
plied to observe the levels of B cell lymphoma/lewkmia-2 (Bcl-2), Bcl associated x protein (Bax), Cytochrome c (Cytc),
cystinyl aspartate specific protease (Caspase-3). Results The neurological behavior score was (0.00±0.00) in sham op-
eration group, (2.38±0.78) in model group, (1.47±0.41) in high dose group, (1.68±0.52) in middle dose group, (2.01±
0.66) in low dose group, which was significantly lower in high dose group and middle dose group than model group (P<
0.05). The apoptotic cell count was (3.37±1.10) in sham operation group, (42.80±3.54) in model group, (28.00±2.28) in
high dose group, (32.40±3.26) in middle dose group, (41.40±1.20) in low dose group, which was significantly lower in
high dose group and middle dose group than model group (P<0.05). The expression levels of apoptosis-related protein
Bax, Cytc, Caspase-3 and Bcl-2 were (3.01±1.12), (2.58±1.74), (2.34±1.37), (65.42±3.65) in sham operation group,
(70.67±3.06), (58.31±5.04), (68.04±5.85), (31.26±2.81) in model group, (40.56±4.52), (33.65±3.44), (41.56±4.52),
(55.64 ± 5.49) in high dose group, (47.29 ± 5.04), (38.09 ± 4.24), (47.29 ± 5.04), (48.33 ± 4.26) in middle dose group,
(53.20±4.70), (44.53±4.39), (53.20±4.70), (40.35±3.17) in low dose group. The high, middle, low dose group could in-
hibit the expression of Bax, Cytc and Caspase-3 protein and increase the expression of Bcl-2 protein, showing statistical-
ly significant difference with model group (P<0.05). The effect of high dose group was the best, and there was a dose-de