核心肽对类风湿关节炎滑膜细胞增殖与凋亡的影响

Effect of core peptide on the proliferation and apoptosis of synoviocytes induced by rheumatoid arthritis

背景:前期研究发现,在类风湿关节炎关节腔中注入核心肽,可以明显抑制病情活跃度且抑制滑膜的浸润增生。但由于T细胞和滑膜细胞之间联系密切,核心肽是通过直接作用于滑膜细胞还是通过T细胞介导抑制滑膜细胞尚缺乏研究。目的:对比观察核心肽对类风湿关节炎和骨性关节炎滑膜细胞增殖与凋亡的影响。方法:采用酶消化法分离类风湿关节炎和骨性关节炎患者手术切取的病变滑膜组织滑膜细胞,MTT法检测滑膜细胞增殖率。取第3代类风湿性关节炎和骨性关节炎滑膜细胞分5组干预:空白对照组、甲氨蝶呤组、10,25,50μmol/L核心肽组。MTT法检测各组滑膜细胞增殖率变化;采用流式细胞仪检测各组滑膜细胞凋亡情况。结果与结论:类风湿关节炎滑膜细胞体外增殖速度较骨性关节炎滑膜细胞快(P<0.05)。3种剂量的核心肽均可抑制骨性关节炎滑膜细胞的增殖,诱导其凋亡,且具有剂量相关性,高剂量时,核心肽更明显表现出对骨性关节炎滑膜细胞凋亡的诱导作用。而对于类风湿关节炎滑膜细胞,只有中、高剂量核心肽显示出明显的诱导细胞凋亡作用,但未见明显的剂量依赖性。核心肽对类风湿关节炎滑膜细胞的凋亡诱导作用明显小于其对骨性关节炎细胞的凋亡诱导作用。结果提示核心肽对滑膜细胞具有一定的直接作用,高剂量核心肽在体外可抑制类风湿关节炎滑膜细胞的过度增殖,而中、高剂量核心肽可诱导类风湿关节炎滑膜细胞的凋亡。

BACKGROUND： Our previous study has identified that the treatment of rheumatoid arthritis （RA） by intraarticular injection of core peptide （CP） significantly suppressed disease activity and inhibited synovial hyperplasia, but the effect of CP which was direct action or through T cell-mediated on synoviocytes did not be still studies. OBJECTIVE： To comparatively examine the effects of CP on the proliferation and apoptosis in synoviocytes of RA and osteoarthritis （OA）. METHODS： Synovial tissues were obtained from RA or OA patients at the time of total joint replacement surgery. Synovial cells were isolated from synovial tissues by collagenase digestion. Cell proliferation was measured with MTT assay. Cells at passages 3 were used for the experiments and randomly divided into five groups： control group, 1 μmol/L MTX, low-dosage CP （10 μmol/L）, middle-dosage CP （25 μmol/L） and high-dosage CP （50 μmol/L） groups. Cell proliferation of each group was assessed by MTT, and apoptotic index of the cells were detected using flow cytometry （FCM）. RESULTS AND CONCLUSION： The proliferation of synoviocytes in RA were found higher than those in OA （P〈0.05）. All three dosage of CP could inhibit the proliferation and induce the apoptosis of synoviocytes in OA, showing a dosage-dependent manner. High-dosage CP could remarkably induce the apoptosis of synoviocytes in OA, but middle or high-dosage CP could remarkably induce the apoptosis of synoviocytes in RA. The effect of CP on synoviocytes in RA was inferior to that on synoviocytes in OA. The results indicated that CP had a direct effect on synoviocytes. High-dosage CP inhibited over-proliferation of synoviocytes in RA, while middle and high-dosage CP might induced apoptosis of synoviocytes in RA.