C.G. Bhakta Institute of Biotechnology, Faculty of Science, Uka Tarsadia University, Bardoli, Surat 394 350, Gujarat, India.
Aura Skin Care, Laxmi Icon 2(nd) Floor, Unai Road, Near Swaminarayan Temple, Vyara 394650, Gujarat, India.
Int Immunopharmacol. 2023 Dec;125(Pt B):111174. doi: 10.1016/j.intimp.2023.111174. Epub 2023 Nov 9.
Generalized vitiligo (GV) is an autoimmune disease characterized by the progressive loss of melanocytes.
Current study was undertaken to assess in-vitro therapeutic potential of Harmine and Kaempferol for GV.
Calcium, calcineurin, NFATC1 levels, cell proliferation were assessed by various kits and ORAI1, PEIZO1, Calcineurin, GSK3B, DYRK1A transcripts and IFN-γ,IL-10,TGF-β protein levels were assessed by qPCR and ELISA in blood and skin biopsy samples from Tregs of 52 patients and 50 controls.
Harmine and Kaempferol treatment enhances Treg suppressive capacity, NFATs and FOXP3 expression in blood and skin Tregs of GV patients (p < 0.05). Furthermore, Harmine and Kaempferol treatment in Tregs increased calcineurin and NFATC1 activity and decreased DYRK1A transcripts in blood and skin Tregs of GV patients(p < 0.05). In-silico analysis revealed that Harmine and Kaempferol might boost Treg suppressive capacity by increasing calcineurin dephosphorylation activity leading to increase NFATs activation and also increase nuclear retention of NFATs by inhibiting DYRK1a phosphorylation activity. Moreover, calcineurin and NFATC1 activity in Tregs were positively correlated with Treg suppressive capacity, NFATC1 and FOXP3 expression (p < 0.05), whereas, DYRK1A transcripts were negatively correlated with Treg suppressive capacity, NFATC1 and FOXP3 expression (p < 0.05). These compounds significantly increased melanocytes' survival and proliferation in Treg:CD4/CD8:SK-Mel-28 cell line co-culture system from GV patients (p < 0.0001).
For the first time the study suggests that Harmine and Kaempferol treated Tregs could control the CD8 and CD4T-cells' proliferation and IFN-γ production, leading to melanocytes' survival and proliferation. These compounds may serve as novel Treg-based therapeutics for GV; however, in vivo studies are warranted to assess the safety and efficacy of these compounds.
泛发性白癜风（GV）是一种以黑色素细胞进行性丧失为特征的自身免疫性疾病。
本研究旨在评估哈梅林和山奈酚对 GV 的体外治疗潜力。
通过各种试剂盒评估钙、钙调神经磷酸酶、NFATC1 水平和细胞增殖，通过 qPCR 和 ELISA 评估血液和皮肤活检样本中转录物（ORAI1、PEIZO1、钙调神经磷酸酶、GSK3B、DYRK1A）和蛋白水平（IFN-γ、IL-10、TGF-β）在 52 名患者和 50 名对照者的 Tregs 中。
哈梅林和山奈酚治疗可增强 GV 患者血液和皮肤 Tregs 的 Treg 抑制能力、NFATs 和 FOXP3 表达（p<0.05）。此外，哈梅林和山奈酚治疗可增加 GV 患者血液和皮肤 Tregs 中的钙调神经磷酸酶和 NFATC1 活性，并降低 DYRK1A 转录物（p<0.05）。计算机分析表明，哈梅林和山奈酚可能通过增加钙调神经磷酸酶去磷酸化活性来增强 Treg 抑制能力，从而增加 NFATs 的激活，同时通过抑制 DYRK1a 磷酸化活性来增加 NFATs 的核保留。此外，Tregs 中的钙调神经磷酸酶和 NFATC1 活性与 Treg 抑制能力、NFATC1 和 FOXP3 表达呈正相关（p<0.05），而 DYRK1A 转录物与 Treg 抑制能力、NFATC1 和 FOXP3 表达呈负相关（p<0.05）。这些化合物在 GV 患者的 Treg：CD4/CD8：SK-Mel-28 细胞系共培养系统中显著增加黑素细胞的存活和增殖（p<0.0001）。
本研究首次表明，哈梅林和山奈酚处理的 Tregs 可控制 CD8 和 CD4T 细胞的增殖和 IFN-γ产生，从而导致黑素细胞的存活和增殖。这些化合物可能成为治疗 GV 的新型基于 Treg 的治疗方法；然而，需要进行体内研究来评估这些化合物的安全性和疗效。
