Pellegrini Carolina, Fornai Matteo, Colucci Rocchina, Tirotta Erika, Blandini Fabio, Levandis Giovanna, Cerri Silvia, Segnani Cristina, Ippolito Chiara, Bernardini Nunzia, Cseri Karolina, Blandizzi Corrado, Haskó György, Antonioli Luca
Department of Clinical and Experimental Medicine, University of Pisa, Via Roma 55, 56126, Pisa, Italy.
Department of Pharmaceutical and Pharmacological Sciences, University of Padova, 35131, Padova, Italy.
J Neuroinflammation. 2016 Jun 13;13(1):146. doi: 10.1186/s12974-016-0608-5.
Parkinson's disease (PD) is frequently associated with gastrointestinal (GI) symptoms, including constipation and defecatory dysfunctions. The mechanisms underlying such disorders are still largely unknown, although the occurrence of a bowel inflammatory condition has been hypothesized. This study examined the impact of central dopaminergic degeneration, induced by intranigral injection of 6-hydroxydopamine (6-OHDA), on distal colonic excitatory tachykininergic motility in rats.
Animals were euthanized 4 and 8 weeks after 6-OHDA injection. Tachykininergic contractions, elicited by electrical stimulation or exogenous substance P (SP), were recorded in vitro from longitudinal muscle colonic preparations. SP, tachykininergic NK1 receptor, and glial fibrillary acidic protein (GFAP) expression, as well as the density of eosinophils and mast cells in the colonic wall, were examined by immunohistochemical analysis. Malondialdehyde (MDA, colorimetric assay), TNF, and IL-1β (ELISA assay) levels were also examined. The polarization of peritoneal macrophages was evaluated by real-time PCR.
In colonic preparations, electrically and SP-evoked tachykininergic contractions were increased in 6-OHDA rats. Immunohistochemistry displayed an increase in SP and GFAP levels in the myenteric plexus, as well as NK1 receptor expression in the colonic muscle layer of 6-OHDA rats. MDA, TNF, and IL-1β levels were increased also in colonic tissues from 6-OHDA rats. In 6-OHDA rats, the number of eosinophils and mast cells was increased as compared with control animals, and peritoneal macrophages polarized towards a pro-inflammatory phenotype.
The results indicate that the induction of central nigrostriatal dopaminergic degeneration is followed by bowel inflammation associated with increased oxidative stress, increase in pro-inflammatory cytokine levels, activation of enteric glia and inflammatory cells, and enhancement of colonic excitatory tachykininergic motility.
帕金森病(PD)常伴有胃肠道(GI)症状,包括便秘和排便功能障碍。尽管有人提出肠道炎症状态的发生,但这些疾病的潜在机制仍大多未知。本研究考察了经黑质内注射6-羟基多巴胺(6-OHDA)诱导的中枢多巴胺能变性对大鼠远端结肠兴奋性速激肽能运动的影响。
在注射6-OHDA后4周和8周对动物实施安乐死。从结肠纵肌制备物中体外记录电刺激或外源性P物质(SP)引发的速激肽能收缩。通过免疫组织化学分析检测SP、速激肽能NK1受体和胶质纤维酸性蛋白(GFAP)的表达,以及结肠壁中嗜酸性粒细胞和肥大细胞的密度。还检测了丙二醛(MDA,比色法)、肿瘤坏死因子(TNF)和白细胞介素-1β(ELISA法)水平。通过实时PCR评估腹膜巨噬细胞的极化。
在结肠制备物中,6-OHDA大鼠电刺激和SP诱发的速激肽能收缩增强。免疫组织化学显示,6-OHDA大鼠肌间神经丛中SP和GFAP水平增加,结肠肌层中NK1受体表达增加。6-OHDA大鼠结肠组织中MDA、TNF和白细胞介素-1β水平也升高。与对照动物相比,6-OHDA大鼠中嗜酸性粒细胞和肥大细胞数量增加,腹膜巨噬细胞向促炎表型极化。
结果表明,中枢黑质纹状体多巴胺能变性的诱导之后会出现肠道炎症,伴有氧化应激增加、促炎细胞因子水平升高、肠胶质细胞和炎症细胞激活以及结肠兴奋性速激肽能运动增强。