Front Immunol:类风湿性关节炎滑液中基质金属蛋白酶-9/明胶酶B的研究

2022-01-11 从医路漫漫 MedSci原创

基础研究与临床研究相结合,使风湿病研究突飞猛进。在诊断水平上,RA抗瓜氨酸肽抗体的发现是另一个突破。瓜氨酸是RA特异性自身抗体抗原决定因素的重要组成部分。

 

研究背景:基础研究与临床研究相结合,使风湿病研究突飞猛进。在诊断水平上,RA抗瓜氨酸肽抗体的发现是另一个突破。瓜氨酸是RA特异性自身抗体抗原决定因素的重要组成部分。检测循环中抗瓜氨酸肽(CCP)的抗体滴度,例如抗瓜氨酸肽抗体(ACPA),用于各种类型关节炎患者的鉴别诊断、治疗随访和预后。

  基质金属蛋白酶(MMPs)在关节炎中的作用也被广泛研究。开放性的研究也可以追溯到1987年,当时Okada和同事描述了MMP-1/成纤维细胞胶原酶、MMP-2/明胶酶A,以及最重要的MMP-3/基质溶蛋白-1与关节炎疾病的关系。MMP-3,以及MMP-1和MMP-2,能够破坏软骨。在抗体诱导的关节炎动物模型中,MMP-2缺乏的小鼠增加了关节炎评分,而MMP -9缺乏的小鼠降低了关节炎评分。表明了MMP-2的保护作用和MMP-9的有害作用。在胶原诱导的小鼠关节炎中,抑制MT1-MMP可与抗TNF治疗协同延缓疾病进展。除了增加ACPA水平外,MMPs水平,特别是MMP-3水平,在关节炎患者的血清和滑膜液中增加,在血浆/血清或滑膜液中检测它们可分别用于诊断系统性和局部疾病。基质金属蛋白酶的直接功能为关节破坏,胶原溶解和蛋白聚糖的降解,基质金属蛋白酶的间接影响,导致自身免疫过程破坏,通过识别中性粒细胞MMP-9 /白明胶酶B,以及II型胶原糖基化残余表位的发现,以评估MMP抑制剂对自身免疫性疾病的治疗效果。

  目的:探讨明胶酶特别是明胶酶B/MMP-9的翻译后修饰(PTMs),包括明胶酶的蛋白水解、多聚化、复合体形成和瓜氨酸化,并检测明胶-琼脂糖亲和纯化的滑膜液中是否存在与关节炎相关的特异性MMP蛋白。

  方法:采用酶谱分析方法检测潜在的、激活的、复杂的和截断的明胶酶A /MMP-2和明胶酶B/MMP-9的蛋白形态,以比较其特异性水平,以底物转换分析方法检测净蛋白水解活性,并通过Western blot分析截断的蛋白变体。通过使用一种新的改良瓜氨酸的单克隆抗体,使得瓜氨酸化检测的灵敏度增强。

  结果:用酶谱分析鉴定了滑液中所有MMP-9和MMP-2的蛋白形式,并对包括类风湿关节炎(RA)在内的各种关节炎疾病中MMP-9和MMP-2的水平进行了研究。其次,我们解决了对MMP-9水平与蛋白水解活性的误解。第三,在存档的RA滑膜液样本中发现了瓜氨酸化的截短的MMP-9蛋白,并在一个小的前瞻性RA样本队列中证实了它的存在。

结论:类风湿关节炎滑液中含有较高水平的MMP-9,包括截短型和瓜氨酸型蛋白。将基质金属蛋白酶-9(MMP-9)作为分析物与其瓜氨酸化的PTM相结合,可能具有临床应用价值,特别是在关节炎领域。

原文出处:Grillet B,  Yu K,  Ugarte-Berzal E,  Janssens R,et al.Proteoform Analysis of Matrix Metalloproteinase-9/Gelatinase B and Discovery of Its Citrullination in Rheumatoid Arthritis Synovial Fluids.Front Immunol 2021;12

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    2022-02-10 晓辰
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    2022-08-15 canlab
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    2022-01-10 lmm397
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