Neuroprotective and neuroregenerative effects of trophic factors

1.MANF：我们首先研究了MANF和甲基苯丙胺(MA)在含有多巴胺能神经元的原代中脑腹侧培养中的相互作用。我们的初步数据表明，MANF减轻了培养的多巴胺能神经元中MA的毒性。我们还发现，MANF可减轻缺血性脑损伤，促进行为恢复。卒中前给予MANF蛋白，剂量为6g，可减少卒中大鼠的脑梗塞，抑制DNA片段化，促进运动恢复。局部应用AAV-MANF可增强大脑皮层神经元和胶质细胞MANF的表达。AAV-MANF可缩小卒中大鼠脑梗塞体积，促进行为恢复。我们的数据表明，给予MANF蛋白或AAV-MANF均可减少缺血性脑损伤。 2.CDNF：我们检测了CDNF注射到C57/BL6小鼠纹状体是否对注射MPTP后的黑质纹状体多巴胺系统具有神经保护和神经修复作用。我们发现，在MPTP暴露前20小时注射双侧纹状体CDNF，在2周后测量时，可以改善水平和垂直运动行为。此外，CDNF预处理增加了纹状体和黑质网状部(SNPr)的酪氨酸羟化酶(TH)免疫阳性反应，以及黑质致密部(SNPC)TH阳性细胞的数量。在MPTP注射后1周给予CDNF后，小鼠的水平和垂直行为增加。此外，注射CDNF后，纹状体多巴胺纤维密度和SNPC内TH阳性细胞数均增加。我们的结论是，在MPTP模型中，纹状体内给予CDNF对黑质纹状体多巴胺系统既有神经保护作用，又有神经修复作用，这支持了基于CDNF的帕金森病治疗策略的发展。 3.BMP7：在原代培养的多巴胺能神经元中，BMP7可降低MA介导的毒性(即降低TH免疫反应性和增加TUNEL标记)。Bmp7还可降低MA介导的体内毒性。脑室注射BMP7可拮抗MA引起的小鼠纹状体TH免疫反应和运动活动的改变。RT-PCR检测发现，侧脑室注射9-顺式维甲酸(9cRA)可增强BMP7mRNA的表达。9cRA可减轻大剂量MA后纹状体TH免疫反应的丧失。在卒中大鼠中，9cRA预处理增加了卒中后2天的运动活动，并减轻了神经功能缺失。9cRA还可减少脑梗塞和TUNEL标记。注射9cRA后1d给予BMP拮抗剂noggin可拮抗上述保护反应。综上所述，我们的数据表明9cRA具有保护作用，这些作用涉及BMP。

1. MANF: We first examined the interaction of MANF and methamphetamine (MA) in primary ventral mesencephalic cultures containing dopaminergic neurons. Our preliminary data indicate that MANF attenuated MA toxicity in dopaminergic neurons in culture. We also found that MANF reduced ischemic brain injury and promoted behavioral recovery. Pre-stroke delivery of MANF protein, at a dose of 6 g, reduced cerebral infarction, suppressed DNA fragmentation, and facilitated motor recovery in stroke rats. Local administration of AAV-MANF enhanced MANF expression in neurons and glia in cerebral cortex. Pretreatment with AAV-MANF reduced the volume of cerebral infarction and facilitated behavioral recovery in stroke rats. Our data suggest that administration of either MANF protein or AAV-MANF reduces ischemic brain injury. 2. CDNF: we examined whether CDNF injections into striatum of C57/Bl6 mice have neuroprotective and neurorestorative properties for the nigrostriatal dopamine system after MPTP injections. We found that bilateral striatal CDNF injections, given 20-h before MPTP exposure, improved horizontal and vertical motor behavior when measured 2 weeks afterwards. In addition, CDNF pre-treatment increased tyrosine hydroxylase (TH)-immunoreactivity in the striatum and in the substantia nigra pars reticulata (SNpr), as well as number of TH-positive cells in substantia nigra pars compacta (SNpc). Post-treatment with CDNF, given 1 week after MPTP injections, increased horizontal and vertical behavior of mice. Furthermore, dopamine fiber densities in striatum and the number of TH positive cells in SNpc were increased after CDNF injections. We conclude that intrastriatal CDNF administration is both neuroprotective and neurorestorative for the nigrostriatal dopamine system in the MPTP model, which supports the development of CDNF-based treatment strategies for Parkinsons disease. 3. BMP7: In primary dopaminergic neuronal culture, BMP7 reduced MA mediated toxicity (i.e. decreased TH immunoreactivity and increasing TUNEL labeling). BMP7 also reduced MA mediated toxicity in vivo. Intra-cerebroventricular administration of BMP7 antagonized MA-induced changes in TH immunoreactivity in striatum and locomotor activity in mice. We found that intracerebroventricular administration of 9-cis-retinoic acid (9cRA) enhanced BMP7 mRNA expression, detected by RTPCR. Pretreatment with 9cRA attenuated the loss of TH immunoreactivity in striatum after high dose of MA administration. In stroke rats, pretreatment with 9cRA increased locomotor activity and attenuated neurological deficits 2 days after stroke. 9cRA also reduced cerebral infarction and TUNEL labeling. These protective responses were antagonized by BMP antagonist noggin given at one day after 9cRA injection. Taken together, our data suggest that 9cRA has protective effects and these effects involve BMPs.