兰尼定受体-1在骨骼肌缺血再灌注损伤中的表达及意义

发布时间：2018-05-07 02:44

本文选题：骨骼肌 + 缺血-再灌注损伤　；参考：《中南大学》2007年硕士论文

【摘要】： 背景和目的缺血组织在恢复血流后出现的更为严重的损伤称之为缺血再灌注损伤(ischemic reperfusion，I／R)。缺血再灌注损伤在急诊外科常见，严重影响患者的功能恢复，甚至造成残疾或死亡。良好的血液循环使组织细胞获得充足的氧和营养物质供应并排出代谢产物，各种原因造成血液供给中断，灌注量减少，都会引起组织和细胞的缺血性损伤，其机制十分复杂。近年来发现：它是一个多因素参与，多机制损伤的过程，普遍认为：白细胞介导的炎症反应，能量代谢障碍，氧自由基，钙超载在其发生过程中起关键作用。本研究旨在讨论钙释放通道兰尼定受体-1(ryanodine receptors 1，RyR_1)在骨骼肌缺血再灌注损伤中的表达及其意义，为阐明其在钙超载中的作用做初步研究。材料和方法健康雄性SD大鼠随机分组，建立后肢骨骼肌缺血再灌注损伤(skeletal muscle ischemia reperfusion injury，SMIRI)动物模型，，运用逆转录—聚合酶链反应(RT-PCR)检测大鼠骨骼肌细胞RyR_1mRNA的表达，运用软件分析其相对浓度。运用比色法测定血清丙二醛(MDA)、一氧化氮(NO)、超氧化歧化酶(SOD)、髓过氧化物酶(MPO)水平，骨骼肌细胞中丙二醛(MDA)、超氧化歧化酶(SOD)、髓过氧化物酶(MPO)，以及湿重／干重(W／D)比例反映骨骼肌水肿程度，光镜观察骨骼肌结构变化。结果骨骼肌缺血再灌注导致RyRImRNA表达在再灌注两小时增强，而后明显降低(P＜0.05)，具有统计学意义。骨骼肌缺血再灌注导致MDA、NO、MPO以及湿重／干重比值明显升高，SOD降低，具有统计学意义(P＜0.05)。结论本研究首先应用RT-PCR方法检测SMIRI时RyR_1mRNA的表达，有助于进一步阐明SMIRI的发生机制。骨骼肌在缺血再灌注后由于多种机制参与，导致骨骼肌细胞RyR_1蛋白表达异常，细胞内钙超载，加上组织缺血时产生的自由基，共同对肢体骨骼肌组织造成一定的损害，为寻找防治SMIRI开辟一条可行性方法奠定了初步基础。
[Abstract]:Background and objective the more serious injury of ischemic tissue after recovery of blood flow is called ischemic reperfusion injury. Ischemia-reperfusion injury is common in emergency surgery, which seriously affects the functional recovery of patients and even results in disability or death. Good blood circulation can make the tissue and cells get sufficient oxygen and nutrients supply and excrete the metabolites. All kinds of reasons lead to the interruption of blood supply and the decrease of perfusion which will cause ischemic injury of tissues and cells. The mechanism is very complex. In recent years, it has been found that it is a multi-factor and multi-mechanism process of injury. It is generally believed that leukocyte mediated inflammation, energy metabolism disorders, oxygen free radicals and calcium overload play a key role in the process of its occurrence. The purpose of this study was to investigate the expression and significance of the calcium release channel Lainidine receptors 1 RyR1 in skeletal muscle ischemia-reperfusion injury, and to elucidate its role in calcium overload. Materials and methods healthy male Sprague-Dawley rats were randomly divided into two groups to establish an animal model of skeletal muscle ischemia / reperfusion injury. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of RyR_1mRNA in skeletal muscle cells of rats. The relative concentration was analyzed by software. The serum levels of malondialdehyde (MDA), nitric oxide (no), superoxide dismutase (SOD) and myeloperoxidase (MPO) were measured by colorimetry. The ratio of malondialdehyde (MDA), superoxide dismutase (SOD), myeloperoxidase (MPO) and wet / dry weight (W / D) in skeletal muscle cells reflected the degree of skeletal muscle edema. The changes of skeletal muscle structure were observed under light microscope. Results the expression of RyRImRNA increased after reperfusion for two hours and then decreased significantly (P < 0.05). MDA-NON-MPO and wet / dry weight ratio increased and SOD decreased significantly (P < 0.05) after ischemia and reperfusion of skeletal muscle. Conclusion in this study, the expression of RyR_1mRNA in SMIRI was detected by RT-PCR method, which was helpful to elucidate the pathogenesis of SMIRI. After ischemia / reperfusion, skeletal muscle has been involved in many mechanisms, which lead to abnormal expression of RyR_1 protein in skeletal muscle cells, intracellular calcium overload, and free radicals produced by tissue ischemia, which cause certain damage to skeletal muscle tissue of limbs. It lays a preliminary foundation for finding a feasible method to prevent and cure SMIRI.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R363

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