论文部分内容阅读
目的研究不同剂量丁酸钠对脑缺血再灌注损伤急性期小鼠炎症反应、神经元凋亡保护机制。方法选取由ICR小鼠124只,将小鼠随机分为4组(每组31只):假手术组、模型组、低剂量实验组(丁酸钠5 mg·kg~(-1))和高剂量试验组(丁酸钠10 mg·kg~(-1));依据改良Zea-Longa线栓法制备大鼠大脑中动脉栓塞模型,血流阻断2 h后拔出线栓,形成再灌注。假手术组小鼠双侧的颈总动脉暴露但血管不进行结扎,表皮的切口正常的缝合,在24 h以后处死小鼠;模型组、低剂量试验组和高剂量试验组在假手术组基础上,结扎双侧颈总动脉,30 min以后解开结扎线,再灌注2 h后灌胃处理,24 h后处死小鼠。用WST-1检测超氧化物歧化酶(SOD)活力,用硫代巴比妥酸检测丙二醛(MDA)含量,用Western blot检测B淋巴细胞癌-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、丝氨酸/苏氨酸激酶(Akt)和磷酸化Akt(p-Akt)蛋白表达量,用ELISA法检测小鼠脑组织内白细胞介素8(IL-8)、白细胞介素1β(IL~(-1)β)和肿瘤坏死因子-α(TNF-α)的含量。结果模型组、假手术组与高剂量试验组SOD为(115.46±25.07),(153.65±34.81),(146.88±15.63)U·mg~(-1),MDA为(2.13±0.27),(1.42±0.07),(1.43±0.08)nmol·mg~(-1),IL~(-1)β分别为(17.82±3.60),(7.64±1.14),(10.15±2.05)pg·mg~(-1);TNF-α分别为(16.03±1.67),(7.95±0.85),(11.08±0.83)pg·mg~(-1),IL-8分别为(15.03±2.66),(9.52±1.05),(10.28±1.17)pg·mg~(-1);Bax/Bcl-2比值分别为0.67±0.05,1.00±0.00,1.13±0.10;p-Akt/Akt比值分别为0.72±0.06,1.00±0.00,1.18±0.12,模型组分别与假手术组和高剂量试验组比较,差异均有统计学意义(均P<0.05)。结论丁酸钠能够抑制炎性因子与氧化应激,使Bcl-2蛋白的表达增加,Bax蛋白的表达减少,从而对神经元的凋亡起到抑制作用。
Objective To study the protective effects of different doses of sodium butyrate on inflammatory reaction and neuronal apoptosis in mice with acute cerebral ischemia-reperfusion injury. Methods A total of 124 ICR mice were selected and randomly divided into 4 groups (31 in each group): sham operation group, model group, low dose group (sodium butyrate 5 mg · kg -1) and High-dose test group (sodium butyrate 10 mg · kg -1); middle cerebral artery occlusion model was established by modified Zea-Longa method; perfusion. Sham-operation group mice bilateral common carotid artery exposed but not ligation of blood vessels, the skin of the incision suture normal, mice were sacrificed after 24 h; model group, low-dose test group and high-dose test group in sham-operated group The common carotid arteries were ligated and the ligature was removed after 30 minutes. The rats were sacrificed 24 hours later after 2 hours of reperfusion. The activity of superoxide dismutase (SOD) was detected by WST-1, MDA content was detected by thiobarbituric acid, Bcl-2, Bcl-2 The expressions of Bax, Akt and p-Akt in the brain tissue were detected by ELISA. The levels of interleukin-8 (IL-8), interleukin 1β (IL-β) and tumor necrosis factor-α (TNF-α) were measured. Results The SOD of model group, sham operation group and high dose group were (115.46 ± 25.07), (153.65 ± 34.81), (146.88 ± 15.63) U · mg -1, MDA was (2.13 ± 0.27) ± 0.07), (1.43 ± 0.08) nmol · mg -1 and IL -1 β were (17.82 ± 3.60), (7.64 ± 1.14), (10.15 ± 2.05) pg · mg ~ (-1) 1 and TNF-α were (16.03 ± 1.67), (7.95 ± 0.85) and (11.08 ± 0.83) pg · mg -1, respectively, and those of IL-8 were 15.03 ± 2.66 and 9.52 ± 1.05, , (10.28 ± 1.17) pg · mg -1; Bax / Bcl-2 ratios were 0.67 ± 0.05, 1.00 ± 0.00 and 1.13 ± 0.10, respectively; p-Akt / Akt ratios were 0.72 ± 0.06 and 1.00 ± 0.00 , 1.18 ± 0.12 respectively. There was significant difference between the model group and sham operation group and high dose test group (all P <0.05). Conclusion Sodium butyrate can inhibit the inflammatory factors and oxidative stress, increase the expression of Bcl-2 protein and decrease the expression of Bax protein, which can inhibit the neuronal apoptosis.