Wang Xiaohong, Chi Jinghong, Huang Di, Ding Li, Zhao Xiaojing, Jiang Lai, Yu Yang, Gao Feng
The 5th Department of Neurology, the Third Affiliated Hospital of Qiqihar Medical University, Qiqihar, Heilongjiang 161000, P.R. China.
The 8th Department of Neurology, the First Hospital of Qiqihar City, Qiqihar, Heilongjiang 161000, P.R. China.
Exp Ther Med. 2020 Feb;19(2):931-938. doi: 10.3892/etm.2019.8297. Epub 2019 Dec 5.
Mechanism by which α-synuclein affects the progression of Parkinson's disease through Pyrin Domain Containing Protein 3 (NLRP3) was explored. Peripheral blood plasma of 40 Parkinson's disease patients and 40 normal healthy people attending the department of neurology of the Third Affiliated Hospital of Qiqihar Medical University were collected from March 2018 to January 2019. The expression levels of oligomers, phosphorylated α-synuclein, interleukin-1β (IL-1β), interleukin-6 (IL-6) and transforming growth factor-α (TGF-α) in plasma were detected by ELISA. Astrocytes in mouse brain tissues were extracted by primary culture method, the cells were divided into drug group and the drug + inhibitor group. After adding 0, 5, 10 and 20 µg oligomerized α-synuclein or 5 mM autophagy inhibitor 3-Methyladenine (3-MA), the expression level of NLRP3, caspase-1, IL-1β and Atg5 proteins in the cells was detected. The expression level of IL-1β in peripheral blood of PD patients was significantly increased (0.604±0.136 µmol/l vs. 1.876±0.327 µmol/l, P=0.002), while there was no significant difference between IL-6 and TGF-α. Both oligomers (0.171±0.045 µmol/l vs. 0.676±0.084 µmol/l, P<0.0001) and phosphorylated α-synuclein (0.128±0.041 µmol/l vs. 0.849±0.108 µmol/l, P<0.0001) in peripheral blood of PD patients were significantly elevated. The expression levels of NLRP3, caspase-1 and IL-1β in mouse astrocytes all increased with the increase of the concentration of oligomerized α-synuclein, and Atg5 protein expression also increased gradually with the concentration, and reached the highest level when the concentration was 10 µg/ml. The expression levels of NLRP3, caspase-1 and IL-1β were inhibited after the addition of autophagy inhibitor 3-MA. α-synuclein mediates the activation of NLRP3 inflammasome in PD patients by upregulating Atg5 protein expression.
探讨了α-突触核蛋白通过含吡啉结构域蛋白3（NLRP3）影响帕金森病进展的机制。于2018年3月至2019年1月收集齐齐哈尔医学院第三附属医院神经内科就诊的40例帕金森病患者及40例正常健康人的外周血血浆。采用酶联免疫吸附测定法检测血浆中寡聚体、磷酸化α-突触核蛋白、白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）及转化生长因子-α（TGF-α）的表达水平。采用原代培养法提取小鼠脑组织中的星形胶质细胞，将细胞分为药物组和药物+抑制剂组。分别加入0、5、10及20μg寡聚化α-突触核蛋白或5 mM自噬抑制剂3-甲基腺嘌呤（3-MA）后，检测细胞中NLRP3、半胱天冬酶-1、IL-1β及自噬相关蛋白5（Atg5）的表达水平。帕金森病患者外周血中IL-1β的表达水平显著升高（0.604±0.136μmol/L比1.876±0.327μmol/L，P = 0.002），而IL-6和TGF-α之间无显著差异。帕金森病患者外周血中寡聚体（0.171±0.045μmol/L比0.676±0.084μmol/L，P<0.0001）和磷酸化α-突触核蛋白（0.128±0.041μmol/L比0.849±0.108μmol/L，P<0.0001）均显著升高。小鼠星形胶质细胞中NLRP3、半胱天冬酶-1及IL-1β的表达水平均随寡聚化α-突触核蛋白浓度的增加而升高，Atg5蛋白表达也随浓度逐渐增加，在浓度为10μg/ml时达到最高水平。加入自噬抑制剂3-MA后，NLRP3、半胱天冬酶-1及IL-1β的表达水平受到抑制。α-突触核蛋白通过上调Atg5蛋白表达介导帕金森病患者NLRP3炎性小体的激活。
