人参皂甙Rd促进成年大鼠脑梗死后神经发生的研究

发布时间：2018-05-16 19:11

本文选题：人参皂甙Rd + 脑梗死　；参考：《第四军医大学》2013年硕士论文

【摘要】：脑卒中具有发病率高、死亡率高和致残率高的特点，严重威胁和影响着人类的健康和人们的生活质量。我国卫生部在2008年公布的第三次全国居民死因调查结果中显示：脑卒中已经上升为我国居民死亡原因的第一位，其中缺血性脑卒中（脑梗死）约占全部脑卒中的80%以上，有效的治疗是挽救患者生命、提高其生活质量的关键，但受限于发病机制的复杂性，其治疗一直是神经病学的重难点所在。截至目前，脑卒中后的治疗有多种方法，唯一有效的溶栓治疗则受限于其过窄的治疗时间窗和对出血等并发症的顾忌，以及应用时严格的适应症和众多禁忌症以及其他客观原因，未能在临床得到广泛应用；神经保护治疗方法经过多年的研究和探索，也没有在临床试验中取得成功。这使得研究人员开始在卒中的急性期和亚急性期内寻求行之有效的神经修复疗法。而在脑缺血后，神经功能缺损即使在不给予药物治疗的情况下，也能够有一定程度的自我恢复，这可能与脑缺血后内源性神经发生的增加有关。因此，通过促进内源性神经发生，，从而改善脑缺血后的神经功能缺损症状，恢复神经功能，成为当前脑卒中后神经修复治疗的重要研究方向。人参皂甙Rd（GSRd）是具有完全自主知识产权的国家一类候选新药。在以我院神经内科牵头的多中心、随机、双盲、安慰剂对照的临床试验结果中首次证实GSRd作为神经保护剂的临床有效性。基础研究方面，我们发现，脑梗死急性期（4小时）给予GSRd能够减少大鼠梗死体积，促进运动功能的恢复，作用机制可能与抑制氧化应激损伤，维持线粒体膜电位有关。近期我们发现在脑梗死后的亚急性期（4-24小时）给予GSRd也能明显促进大鼠神经功能恢复；同时，在动物实验和细胞培养中发现：GSRd能够促进成年大鼠SVZ和SGZ的神经干细胞/前体细胞的增殖、促进体外培养的神经干细胞向神经元分化。鉴于此，我们推测：GSRd能明显改善脑梗死亚急性期预后的机制可能与促进神经发生有关；如果能够证实这个假设，将具有非常重要的临床应用价值，成为脑梗死后治疗的突破性进展。目的：利用大鼠脑缺血/再灌注模型研究脑梗死后不同时间点给予GSRd对SVZ区神经发生的影响。方法：（1）成年SD雄性大鼠，线栓法建立短暂性局限性大鼠大脑中动脉栓塞/再灌注（MCAO/R）模型，假手术组方法同模型组，但不阻断MCA血供；通过神经功能评分和TTC染色观察动物脑梗死模型的成功率和梗死体积的稳定性。（2）MCAO/R和假手术大鼠在脑梗死后的6h、24h或3d腹腔注射GSRd（剂量为10mg/kg/d，共7d），对照组（模型组和假手术组）腹腔注射等体积GSRd专用溶剂；在用药后即刻、1w、2w和3w灌注取材，进行免疫组织化学染色，观察SVZ区NSCs的增殖、迁移及分化情况，探讨GSRd与神经发生的关系。结果：（1）线栓法建立大脑中动脉栓塞/再灌注模型，梗死体积稳定，成功率和长期存活率较高。（2）在脑梗死后6h或24h给药，与对照组比较，GSRd可显著促进NSCs增殖和迁移，且随着时间延长逐渐减弱，对分化的作用不明显；脑梗死后第3d给予GSRd，对NSCs的增殖、迁移和分化情况与对照组之间无差异。结论：在脑梗死后24h内给予GSRd可促进SVZ区的NSCs增殖，但不影响其迁移和分化。
[Abstract]:Stroke has the characteristics of high incidence, high mortality and high disability rate, which seriously threaten and affect human health and people's quality of life. The results of the third National Death survey published by the Ministry of health in 2008 showed that stroke has risen to be the first cause of death in our country, including ischemic stroke. (cerebral infarction) accounts for more than 80% of all stroke. Effective treatment is the key to save the life of the patient and improve the quality of life. But limited to the complexity of the pathogenesis, the treatment has been the most difficult problem in neurology. At present, there are many methods for the treatment after stroke, the only effective thrombolytic therapy is limited by its too narrow. The time window for the treatment and the scruples of complications such as bleeding, as well as the strict indications and many contraindications and other objective reasons in the application are not widely used in clinical practice; the neuroprotective therapy has not been successful in clinical trials after years of research and exploration. This makes the researchers start in the stroke. In the acute and subacute phase, an effective nerve repair therapy is sought. After cerebral ischemia, the neural function defect can also have a certain degree of self recovery, even in the absence of drug treatment, which may be related to the increase of endogenous neurogenesis after cerebral ischemia. Therefore, the endogenous neurogenesis can be improved by promoting endogenous neurogenesis. It is an important research direction of neural repair after stroke that good symptoms and neurological functions after cerebral ischemia.
Ginsenoside Rd (GSRd) is a national candidate drug with fully autonomous intellectual property rights. The clinical effectiveness of GSRd as a neuroprotective agent is first confirmed in the results of a multicenter, double-blind, placebo-controlled clinical trial, led by the neurology department of our hospital, and we have found that the acute phase of cerebral infarction (4 hours) is given to G. SRd can reduce the infarct volume and promote the recovery of motor function. The mechanism of action may be related to the inhibition of oxidative stress injury and the maintenance of mitochondrial membrane potential. In the near future, we found that the subacute phase (4-24 hours) after cerebral infarction (GSRd) can also significantly promote the recovery of neural function in rats; at the same time, in animal experiments and cell culture. Now: GSRd can promote the proliferation of neural stem cells / precursor cells of SVZ and SGZ in adult rats and promote the differentiation of neural stem cells from neurons in vitro. In view of this, we speculate that the mechanism of GSRd can obviously improve the mechanism of prognosis in the subacute phase of cerebral infarction and may be related to the promotion of neurogenesis. The important clinical application value has become a breakthrough in the treatment of cerebral infarction.
Objective: To study the effects of GSRd on the neurogenesis in the SVZ area at different time points after cerebral infarction in rats.
Methods: (1) Adult SD male rats were used to establish a transient localized rat model of middle cerebral artery embolism / reperfusion (MCAO/R), the sham operation group was the same as the model group, but did not block the MCA blood supply. The success rate and the stability of the infarct volume were observed by the neural function score and TTC staining. (2) MCAO/R and the sham operation. Rats were intraperitoneally injected with GSRd (10mg/kg/d, total 7D) in 6h, 24h or 3D after cerebral infarction. The control group (model group and sham operation group) was intraperitoneally injected with equal volume of GSRd specific solvent, and immediately after medication, 1W, 2W and 3W perfusion were obtained by immunohistochemistry. The proliferation, migration and differentiation of SVZ District NSCs were observed and the neurogenesis and neurogenesis were observed. The relationship.
Results: (1) the embolic and reperfusion model of middle cerebral artery was established by the method of thread embolism. The infarct volume was stable, the success rate and the long-term survival rate were higher. (2) 6h or 24h was given after cerebral infarction. Compared with the control group, GSRd could significantly promote the proliferation and migration of NSCs, and the effect on differentiation was not obvious with the time prolonged; 3D was given to GSR after cerebral infarction. D, there was no difference in the proliferation, migration and differentiation of NSCs between the control group and the control group.
Conclusion: GSRd in 24h after cerebral infarction can promote the proliferation of NSCs in SVZ region, but it does not affect its migration and differentiation.
【学位授予单位】：第四军医大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R743.33

【参考文献】