PGC-1α介导线粒体生物发生对骨癌痛伴发镜像痛的影响[*]

The effect of PGC-1α mediates mitochondrial biogenesis on bone cancer pain with concurrent mirror-image pain

摘要:

目的： 探讨PGC-1α调控线粒体生物发生对骨癌痛伴发镜像痛的影响。方法：第一部分实验中大鼠随机分成2组，假手术组（sham组）与骨癌痛组（BCP组）。第二部分实验中大鼠随机分成3组，sham+vehicle（sham）组、BCP+vehicle（溶媒）组、BCP+ZLN005（给药）组。将MRMT-1大鼠乳腺癌细胞注入左胫骨髓腔建立BCP大鼠模型，sham组大鼠注射等体积Hank’s平衡盐溶液。第二部分实验中，给药组大鼠鞘内注射ZLN005（PGC-1α激活剂，单次单剂量100μg/30μL），sham组与溶媒组注射等体积溶剂。Western blot用于分析大鼠脊髓中PGC-1α、Nrf1和Tfam蛋白表达量。qPCR用于分析mtDNA相对拷贝数。免疫荧光双标染色分析PGC-1α的分布与细胞定位。结果： 单侧癌细胞接种诱导大鼠双侧后爪出现机械性痛觉过敏。ZLN005鞘内注射可减轻BCP大鼠双侧的机械性痛觉过敏。在BCP大鼠的脊髓中，观察到PGC-1α、Nrf1、Tfam 和 mtDNA拷贝数水平下降。鞘内注射ZLN005后，PGC-1α、Nrf1、Tfam 和mtDNA拷贝数的表达升高。PGC-1α在同侧和对侧脊髓背角均有分布。PGC-1α的荧光强度在BCP大鼠双侧脊髓中出现下降。PGC-1α主要与神经元共定位。 结论：脊髓中的线粒体生物发生的损伤促进了骨癌痛伴发镜像痛的发生发展。PGC-1α激活介导线粒体生物发生缓解骨癌痛伴发镜像痛现象。

Abstract:

Objective: To investigate the effects and mechanisms of PGC-1α-regulated mitochondrial biogenesis on bone cancer pain with concurrent mirror-image pain. Methods: In the first part of the experiment, the animals were divided into 2 groups, sham group and BCP group. In the second part of the experiment, the animals were divided into 3 groups, sham+vehicle (sham) group, BCP+vehicle (vehicle) group, BCP+ZLN005 (ZLN) group. MRMT-1 breast cancer cells were injected into the medullary cavity of the left tibia to establish BCP rat model, and sham group rats were injected with equal volume of Hank's balanced salt solution. In the second part of the experiment, rats in the ZLN group were injected intrathecally with ZLN005 (PGC-1α activator, single single dose of 100μg/30μL), and the sham group and vehicle group rats were injected with an equal volume of vehicle. The levels of PGC-1α, Nrf1, Tfam were analysed in the spinal cord of the rats by western blot. And mtDNA copy number expression was analysed by qPCR. The localisation of PGC-1α was analysed by immunofluorescence. Results: Mechanical hyperalgesia was induced in the bilateral hind paws of rats by intra-bone marrow injection of cancer cells. ZLN005 intrathecally attenuated mechanical hyperalgesia in BCP rats. In the spinal cord of BCP rats, a decreased in levels of PGC-1α, Nrf1, Tfam and mtDNA copy number were observed. The expression of PGC-1α, Nrf1, Tfam and mtDNA copy numbe was elevated after administration with ZLN005. The distribution of PGC-1α was demonstrated in the spinal dorsal horn on both ipsilateral and contralateral sides. The fluorescence intensity of PGC-1α was declined in BCP rats. PGC-1α was mainly co-located with neurons. Conclusions: In the spinal cord, damage to mitochondrial biogenesis facilitated the development of bone cancer pain concomitant with mirror-image pain. PGC-1α activation can alleviate bone cancer pain with concurrent mirror-image pain by promoting mitochondrial biogenesis.