吸入麻醉药异氟醚对β淀粉样蛋白诱导的大鼠PC12细胞凋亡和内质网应激的影响及其机制
发布时间：2018-11-15 17:46
【摘要】：研究背景：伴随世界老龄化进程的加剧，具有年龄依赖性的阿尔茨海默病已成为21世纪威胁人类健康的重要疾病之一，因其发病机制复杂，发病率、死亡率高，尚无有效治疗措施，其相关领域成为了科学界的研究热点。近年来诸多研究表明蛋白聚集与神经元的损伤及死亡之间有着密切的联系；细胞凋亡与阿尔茨海默病密切相关，但其机制尚未十分清楚，可能与线粒体损伤、氧化应激、钙稳态失衡等因素相关；同时，研究者发现内质网对于机体内环境的改变比较敏感，当细胞受到某些内外因素的刺激时，细胞启动内质网应激机制，但长时间、持续的内质网应激将激活细胞死亡级联反应，进而参与阿尔茨海默病的发病机制和病理过程。临床和动物学研究表明，全身麻醉能够加重阿尔茨海默病的症状和病理过程，，降低其发病年龄，增加其发病率。相关文献报道，临床常用的吸入麻醉药异氟醚可能通过增加脑内异常蛋白的产生和聚集，激活凋亡级联信号通路，诱导神经细胞凋亡，介导阿尔茨海默病的发生。目前全世界每年有2.3亿人在接受麻醉和手术，其中接受麻醉的阿尔茨海默病患者达800万以上，存在巨大的麻醉安全风险和隐患。因此，探究吸入麻醉药异氟醚、阿尔茨海默病与细胞凋亡之间的内在联系具有重要的临床和社会意义。 目的：探讨吸入麻醉药异氟醚对β淀粉样蛋白（A）25-35诱导的大鼠PC12细胞凋亡和内质网应激的影响，并阐明其机制。 方法：将PC12细胞随机分为4组：正常对照组（C组）、A25-3510μmol/L处理组（A组）、2%异氟醚处理组（Iso组）和2%异氟醚和A25-3510μmol/L处理组（Iso+A组）。各组PC12细胞药物处理24h后应用四甲基偶氮唑盐（MTT）比色法检测细胞活性，Hoechst33342核染色法检测PC12细胞凋亡形态，Western blot检测内质网分子伴侣蛋白GRP78及其应激相关凋亡信号蛋白CHOP、JNK和caspase-12表达。 结果：与C组比较，A组和Iso组PC12细胞存活率明显降低（P 0.05），细胞凋亡率明显增加（P 0.05），内质网分子伴侣蛋白GRP78表达明显上调（P 0.05），内质网应激相关凋亡信号蛋白CHOP、p-JNK和caspase-12表达均明显增强（P 0.05）；与A组比较，Iso+A组PC12细胞存活率明显降低（P 0.05），细胞凋亡率明显增加（P 0.05），内质网分子伴侣蛋白GRP78和内质网应激相关凋亡信号蛋白CHOP、p-JNK和caspase-12表达均明显增强（P 0.05）。 结论：异氟醚能够促进Aβ25-35诱导的大鼠PC12细胞凋亡，其机制与激活内质网应激及其相关的凋亡信号通路有关。
[Abstract]:Background: with the aggravation of the aging process in the world, Alzheimer's disease with age dependence has become one of the most important diseases threatening human health in the 21st century, because of its complex pathogenesis, high morbidity and high mortality. There is no effective treatment, and its related field has become a hot spot in the scientific community. In recent years, many studies have shown that there is a close relationship between protein aggregation and neuronal injury and death. Apoptosis is closely related to Alzheimer's disease, but its mechanism is not very clear, which may be related to mitochondrial damage, oxidative stress, calcium homeostasis and other factors. At the same time, the researchers found that endoplasmic reticulum is more sensitive to changes in the body's internal environment. When cells are stimulated by some internal and external factors, the cells activate the endoplasmic reticulum stress mechanism, but for a long time. Persistent endoplasmic reticulum stress activates the cascade of cell death, and then participates in the pathogenesis and pathological process of Alzheimer's disease. Clinical and zoological studies have shown that general anesthesia can aggravate the symptoms and pathological process of Alzheimer's disease, reduce the age of onset and increase the incidence of Alzheimer's disease. It has been reported that inhaled anesthetic isoflurane may activate apoptotic cascade signaling pathway by increasing the production and aggregation of abnormal proteins in the brain, induce neuronal apoptosis and mediate the occurrence of Alzheimer's disease. At present, 230 million people in the world are undergoing anesthesia and surgery every year. Among them, more than 8 million patients with Alzheimer's disease are anesthetized, and there are huge risks and hidden dangers in anesthetic safety. Therefore, it is of great clinical and social significance to explore the relationship between inhaled anesthetic isoflurane, Alzheimer's disease and apoptosis. Aim: to investigate the effects of inhaled anesthetic isoflurane on apoptosis and endoplasmic reticulum stress of rat PC12 cells induced by 尾 -amyloid (A) 25-35, and to elucidate its mechanism. Methods: PC12 cells were randomly divided into 4 groups: normal control group (group C), A25-3510 渭 mol/L group (group A), 2% isoflurane treatment group (Iso group) and 2% isoflurane and A25-3510 渭 mol/L treatment group (Iso A group). The activity of PC12 cells was detected by tetramethylazolium (MTT) colorimetry 24 hours after drug treatment, and the apoptotic morphology of PC12 cells was detected by Hoechst33342 nuclear staining, and the endoplasmic reticulum molecular chaperone protein GRP78 and its stress-related apoptotic signal protein CHOP, were detected by Hoechst33342 nuclear staining. JNK and caspase-12 expression. Results: compared with group C, the survival rate of PC12 cells in group A and Iso was significantly lower than that in group C (P 0.05), the apoptosis rate was significantly increased (P 0.05), and the expression of ER molecular chaperone GRP78 was up-regulated (P 0.05). Endoplasmic reticulum stress-related apoptotic signal proteins (CHOP,p-JNK and caspase-12) were significantly increased (P 0.05). Compared with group A, the survival rate of PC12 cells in, Iso A group was significantly lower than that in group A (P 0.05), and the apoptosis rate was significantly increased (P 0.05). Endoplasmic reticulum molecular chaperone protein (GRP78) and endoplasmic reticulum stress-related apoptotic signal protein (CHOP,) were also significantly increased in, Iso A group. The expression of p-JNK and caspase-12 were significantly increased (P0. 05). Conclusion: isoflurane can promote apoptosis of rat PC12 cells induced by A 尾 25-35, and its mechanism is related to activation of endoplasmic reticulum stress and its related apoptosis signaling pathway.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R614
本文编号：2334003
[Abstract]:Background: with the aggravation of the aging process in the world, Alzheimer's disease with age dependence has become one of the most important diseases threatening human health in the 21st century, because of its complex pathogenesis, high morbidity and high mortality. There is no effective treatment, and its related field has become a hot spot in the scientific community. In recent years, many studies have shown that there is a close relationship between protein aggregation and neuronal injury and death. Apoptosis is closely related to Alzheimer's disease, but its mechanism is not very clear, which may be related to mitochondrial damage, oxidative stress, calcium homeostasis and other factors. At the same time, the researchers found that endoplasmic reticulum is more sensitive to changes in the body's internal environment. When cells are stimulated by some internal and external factors, the cells activate the endoplasmic reticulum stress mechanism, but for a long time. Persistent endoplasmic reticulum stress activates the cascade of cell death, and then participates in the pathogenesis and pathological process of Alzheimer's disease. Clinical and zoological studies have shown that general anesthesia can aggravate the symptoms and pathological process of Alzheimer's disease, reduce the age of onset and increase the incidence of Alzheimer's disease. It has been reported that inhaled anesthetic isoflurane may activate apoptotic cascade signaling pathway by increasing the production and aggregation of abnormal proteins in the brain, induce neuronal apoptosis and mediate the occurrence of Alzheimer's disease. At present, 230 million people in the world are undergoing anesthesia and surgery every year. Among them, more than 8 million patients with Alzheimer's disease are anesthetized, and there are huge risks and hidden dangers in anesthetic safety. Therefore, it is of great clinical and social significance to explore the relationship between inhaled anesthetic isoflurane, Alzheimer's disease and apoptosis. Aim: to investigate the effects of inhaled anesthetic isoflurane on apoptosis and endoplasmic reticulum stress of rat PC12 cells induced by 尾 -amyloid (A) 25-35, and to elucidate its mechanism. Methods: PC12 cells were randomly divided into 4 groups: normal control group (group C), A25-3510 渭 mol/L group (group A), 2% isoflurane treatment group (Iso group) and 2% isoflurane and A25-3510 渭 mol/L treatment group (Iso A group). The activity of PC12 cells was detected by tetramethylazolium (MTT) colorimetry 24 hours after drug treatment, and the apoptotic morphology of PC12 cells was detected by Hoechst33342 nuclear staining, and the endoplasmic reticulum molecular chaperone protein GRP78 and its stress-related apoptotic signal protein CHOP, were detected by Hoechst33342 nuclear staining. JNK and caspase-12 expression. Results: compared with group C, the survival rate of PC12 cells in group A and Iso was significantly lower than that in group C (P 0.05), the apoptosis rate was significantly increased (P 0.05), and the expression of ER molecular chaperone GRP78 was up-regulated (P 0.05). Endoplasmic reticulum stress-related apoptotic signal proteins (CHOP,p-JNK and caspase-12) were significantly increased (P 0.05). Compared with group A, the survival rate of PC12 cells in, Iso A group was significantly lower than that in group A (P 0.05), and the apoptosis rate was significantly increased (P 0.05). Endoplasmic reticulum molecular chaperone protein (GRP78) and endoplasmic reticulum stress-related apoptotic signal protein (CHOP,) were also significantly increased in, Iso A group. The expression of p-JNK and caspase-12 were significantly increased (P0. 05). Conclusion: isoflurane can promote apoptosis of rat PC12 cells induced by A 尾 25-35, and its mechanism is related to activation of endoplasmic reticulum stress and its related apoptosis signaling pathway.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R614
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相关期刊论文 前3条
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2 赵美玲;季宇彬;毕明刚;;细胞凋亡的死亡受体途径[J];黑龙江医药;2013年02期
3 王晓映;刘鹏;朱华;徐艳峰;马春梅;代小伟;刘颖;秦川;;阿尔茨海默病模型小鼠凋亡相关蛋白的表达[J];中国老年学杂志;2012年07期
本文编号：2334003
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