Liang Keshan, Zhang Jingling, Yin Chengbin, Zhou Xueying, Zhou Shengnian
Department of Neurology, Qilu Hospital of Shandong University and Brain Science Research Institute, Shandong University, Jinan, Shandong 250012, P.R. China; Department of Neurology, Pingyi Branch of Qilu Hospital, Shandong University, Pingyi, Shandong 273300, P.R. China.
Department of Endocrinology, Linyi People's Hospital, Linyi, Shandong 276000, P.R. China.
Exp Ther Med. 2017 Feb;13(2):576-580. doi: 10.3892/etm.2016.4006. Epub 2016 Dec 27.
We investigated the protective effects and mechanism of TPX2 on apoptosis of rat neurocytes. A total of 90 SD rats were randomly divided into the drug group, the control group and the blank group, with 30 rats in each group. The rats in the drug group and in the blank group were anesthetized with 10% chloral hydrate (at the dose of 0.5 ml/100 g) and Aβ, with the concentration of 5 µl (1 µg/µl), was injected in the exact position of bilateral hippocampal areas of rats to establish the model. The configured TPX2 inhibitors and edible benne oil were mixed and made into a suspension. After model establishment, the rats were given different treatment methods; the rats in the drug group were given gavage administration in the proportion of 75 mg/kg once a day. The rats in the control group were given intragastric administration with the same proportion of physiological saline once a day. The blank group was the normal healthy group and the rats in this group did not undergo any surgery or drug treatment. Brain tissue in rats were divided into two parts, one part was fixed, dehydrated, paraffin-embedded and made into slices of approximately 5 µm. TUNEL staining was used to examine the apoptosis of brain tissue, H&E staining was used to observe the brain tissue cells of each group, and western blotting for detecting the MAPK, Erk and expression levels of p38 and RT-polymerase chain reaction method was employed to examine mRNA expression levels of MAPK, Erk and p21. After one week, TUNEL staining showed that apoptosis of brain tissue in the drug group was significantly greater than those of the control and blank groups. The protein expression levels of MAPK, Erk and p38 were significantly higher than those of the control group and the normal healthy group; the differences were statistically significant (P<0.05). Western blotting showed that the protein expression levels of MAPK, Erk and p38 of the drug group were significantly lower than those of the control group but higher than those of the normal healthy group; the differences were statistically significant (P<0.05). TPX2 has a protective effect on the apoptosis of brain tissue processed by Aβ, which plays its role through the inhibition of the protein expression levels of MAPK, Erk and p38.
我们研究了TPX2对大鼠神经细胞凋亡的保护作用及机制。将90只SD大鼠随机分为药物组、对照组和空白组，每组30只。药物组和空白组大鼠用10%水合氯醛（剂量为0.5 ml/100 g）麻醉，向大鼠双侧海马区精确位置注射浓度为5 μl（1 μg/μl）的Aβ以建立模型。将配制好的TPX2抑制剂与食用芝麻油混合制成混悬液。造模后，对大鼠给予不同处理方法；药物组大鼠按75 mg/kg的比例每天灌胃给药1次。对照组大鼠每天按相同比例给予生理盐水灌胃。空白组为正常健康组，该组大鼠未接受任何手术或药物治疗。将大鼠脑组织分为两部分，一部分固定、脱水、石蜡包埋并制成约5 µm的切片。采用TUNEL染色检测脑组织凋亡情况，用苏木精-伊红（H&E）染色观察各组脑组织细胞情况，用蛋白质免疫印迹法检测丝裂原活化蛋白激酶（MAPK）、细胞外信号调节激酶（Erk）和p38的表达水平，并用逆转录聚合酶链反应法检测MAPK、Erk和p21的mRNA表达水平。一周后，TUNEL染色显示药物组脑组织凋亡明显高于对照组和空白组。MAPK、Erk和p38的蛋白表达水平明显高于对照组和正常健康组；差异有统计学意义（P<0.05）。蛋白质免疫印迹法显示，药物组MAPK、Erk和p38的蛋白表达水平明显低于对照组，但高于正常健康组；差异有统计学意义（P<0.05）。TPX2对Aβ处理的脑组织凋亡具有保护作用，其通过抑制MAPK、Erk和p38的蛋白表达水平发挥作用。
