摘　要：类风湿关节炎（RA）是一类以滑膜增生、滑膜关节损害、活动功能不全为主要特点的慢性全身性自我免疫炎症性病变。目前，对于RA的治疗包括药物治疗、外科手术治疗和物理疗法。物理疗法中光疗法由于其无侵袭力、无痛等优势，被用作辅助治疗手段普遍应用于临床。为此，本研究通过630 nm LED红光探讨了光生物调节法通过促进成纤维样滑膜细胞凋亡，从而治愈类风湿性关节炎的机理。JC-1染色试验表明，630 nm LED光照后线粒体膜电位去极化增强，膜电位降低；使用CASP3活性检测试剂盒和CASP9活性检测试剂盒进行试验，证明630 nm LED可以促进MH7A细胞中CASP3和CASP9酶的活性；实时荧光定量PCR（RT-qPCR）试验显示，630 nm LED可以促进瞬时受体电位香草素1（TRPV1）的表达；蛋白质免疫印迹试验（Western blot）证明，经过630 nm LED红光照射可以促进MH7A细胞TRPV1蛋白的表达；CCK-8试验表明，630 nm LED可以显著性抑制MH7A细胞活力，但不能降低TNF-α刺激下TRPV1拮抗剂处理的MH7A细胞活性；TUNEL试验证实，630 nm LED照射诱导MH7A细胞凋亡，但不能诱导TNF-α刺激下TRPV1拮抗剂处理的MH7A细胞凋亡。综上所述，630 nm LED红光在类风湿性关节炎中通过调控TRPV1促进TNF-α诱导的MH7A细胞凋亡，这对辅助缓解RA滑膜炎症有一定作用。
关键词：发光二极管；类风湿关节炎；细胞凋亡；成纤维样滑膜细胞；TRPV1
中图分类号：R318.51                           文献标志码：A                DOI：10.3969/j.issn.1007-7146.2022.03.005

Abstract: Rheumatoid arthritis (RA) is a chronic systemic autoimmune inflammatory disease characterized by synovial hyperplasia, synovial joint damage and dysfunction. At present, the treatment of RA mainly includes drug therapy, surgical treatment and physical therapy. In physical therapy, light therapy is used extensively as an adjuvant therapy because of its non-invasive and painless advantages. Therefore, in this study, 630 nm LED red light was used to explore the mechanism of photobiomodulation in the treatment of rheumatoid arthritis by promoting the apoptosis of synovial fibroblasts. The result of JC-1 assay demonstrated that the depolarization of mitochondrial membrane potential increased and the membrane potential decreased after 630 nm LED illumination. Using CASP3 and CASP9 activity detection kit, it is demonstrated that 630 nm LED promoted the activity of the pro-apoptotic CASP3 and CASP9 in synovial fibroblasts. RT-qPCR and Western blot showed that the expression of TRPV1 was promoted by 630 nm LED in MH7A cells. CCK-8 assay showed that the viability of MH7A cells was significantly inhibited by 630 nm LED, whereas the activity of MH7A cells treated with TRPV1 antagonist and TNF-α was not inhibited by 630 nm LED. TUNEL assay showed that apoptosis of MH7A cells was promoted by 630 nm LED, whereas apoptosis of MH7A cells treated with TRPV1 antagonist and TNF-α was not promoted by 630 nm LED. In conclusion, 630 nm LED red light promoted apoptosis by regulating TRPV1 in the TNF-α induced MH7A cells.
Key words: LED; rheumatoid arthritis; apoptosis; fibroblast-like synovial cells; transient receptor potential vanilloid 1
（Acta Laser Biology Sinica, 2022, 31(3): 224-233）