Chang Jui-Chih, Wu Shey-Lin, Liu Ko-Hung, Chen Ya-Hui, Chuang Chieh-Sen, Cheng Fu-Chou, Su Hong-Lin, Wei Yau-Huei, Kuo Shou-Jen, Liu Chin-San
Vascular and Genomic Center, Changhua Christian Hospital, Changhua, Taiwan.
Department of Neurology, Changhua Christian Hospital, Changhua, Taiwan.
Transl Res. 2016 Apr;170:40-56.e3. doi: 10.1016/j.trsl.2015.12.003. Epub 2015 Dec 15.
Although restoration of mitochondrial function in mitochondrial diseases through peptide-mediated allogeneic mitochondrial delivery (PMD) has been demonstrated in vitro, the in vivo therapeutic efficacy of PMD in Parkinson's disease (PD) has yet to be determined. In this study, we compared the functionality of mitochondrial transfer with or without Pep-1 conjugation in neurotoxin (6-hydroxydopamine, 6-OHDA)-induced PC12 cells and PD rat models. We injected mitochondria into the medial forebrain bundle (MFB) of the PD rats after subjecting the nigrostriatal pathway to a unilateral 6-OHDA lesion for 21 days, and we verified the effectiveness of the mitochondrial graft in enhancing mitochondrial function in the soma of the substantia nigra (SN) neuron through mitochondrial transport dynamics in the nigrostriatal circuit. The result demonstrated that only PMD with allogeneic and xenogeneic sources significantly sustained mitochondrial function to resist the neurotoxin-induced oxidative stress and apoptotic death in the rat PC12 cells. The remaining cells exhibited a greater capability of neurite outgrowth. Furthermore, allogeneic and xenogeneic transplantation of peptide-labeled mitochondria after 3 months improved the locomotive activity in the PD rats. This increase was accompanied by a marked decrease in dopaminergic neuron loss in the substantia nigra pars compacta (SNc) and consistent enhancement of tyrosine hydroxylase-positive immunoreaction of dopaminergic neurons in the SNc and striatum. We also observed that in the SN dopaminergic neuron in the treated PD rats, mitochondrial complex I protein and mitochondrial dynamics were restored, thus ameliorating the oxidative DNA damage. Moreover, we determined signal translocation of graft allogeneic mitochondria from the MFB to the calbindin-positive SN neuron, which demonstrated the regulatory role of mitochondrial transport in alleviating 6-OHDA-induced degeneration of dopaminergic neurons.
尽管通过肽介导的同种异体线粒体递送（PMD）恢复线粒体疾病中的线粒体功能已在体外得到证实，但PMD在帕金森病（PD）中的体内治疗效果尚未确定。在本研究中，我们比较了在神经毒素（6-羟基多巴胺，6-OHDA）诱导的PC12细胞和PD大鼠模型中，有或没有Pep-1偶联的线粒体转移的功能。在黑质纹状体通路单侧6-OHDA损伤21天后，我们将线粒体注射到PD大鼠的内侧前脑束（MFB）中，并通过黑质纹状体回路中的线粒体运输动力学，验证了线粒体移植在增强黑质（SN）神经元胞体中线粒体功能方面的有效性。结果表明，只有来自同种异体和异种来源的PMD能显著维持线粒体功能，以抵抗神经毒素诱导的大鼠PC12细胞中的氧化应激和凋亡死亡。其余细胞表现出更强的神经突生长能力。此外，3个月后肽标记的线粒体的同种异体和异种移植改善了PD大鼠的运动活性。这种增加伴随着黑质致密部（SNc）中多巴胺能神经元损失的显著减少，以及SNc和纹状体中多巴胺能神经元酪氨酸羟化酶阳性免疫反应的持续增强。我们还观察到，在接受治疗的PD大鼠的SN多巴胺能神经元中，线粒体复合物I蛋白和线粒体动力学得到恢复，从而改善了氧化性DNA损伤。此外，我们确定了移植的同种异体线粒体从MFB到钙结合蛋白阳性SN神经元的信号转位，这证明了线粒体运输在减轻6-OHDA诱导的多巴胺能神经元变性中的调节作用。
