Wu Xiao, Liu Xuguang, Jing Zhongkun, Chen Yang, Liu Huahui, Ma Wenbin
Chengdu University of Traditional Chinese Medicine 37 Shi'er Qiao Road, Jinniu District, Chengdu 610072, Sichuan, People's Republic of China.
Acupuncture Department of Chinese Medicine Hospital Affiliated to Southwest Medical University 182 Chunhui Road, Longmatan District, Luzhou 646000, Sichuan, People's Republic of China.
Am J Transl Res. 2020 May 15;12(5):1459-1468. eCollection 2020.
The key clinical symptoms and previous findings of RA show a circadian variation, with more prominent joint swelling, stiffness, and pain occurring in the early morning. Moxibustion is able to relieve RA in various pass ways, however, there is no verifying study results for the pathological rhythm of RA. Therefore, we conducted this work to verify whether moxibustion could adjust RA circadian rhythm according to regulate core clock genes. Based on these previous findings that circadian timekeeping is disturbed in RA at molecular level, the aim of this study was to observe the influence of moxibustion on expression level and circadian rhythm of REV-ERBα at different tissues of RA rats. Furthermore, the expression level of core clock genes closely related to RA were evaluated by RT-PCR. 96 SD rats were randomly assigned as 1:1:1:1 ratio to 4 groups for normal control group, RA model group, 5-7 am moxibustion group, and 5-7 pm moxibustion group. RT-PCR was used to measure the relatively expression quantity of REV-ERBα, CLOCK, BMAL1, and PER2 in hypothalamus, hippocampus, and adrenal gland. In RA rats, the expression level of REV-ERBα mRNA were up-regulated in different tissues, and moxibustion potentially up-regulated them in different degrees. In untreated RA rats, the circadian rhythm of REV-ERBα mRNA in hippocampus and adrenal gland both disappeared (P>0.05) and moxibustion was able to recover them (P<0.05). The expression level of CLOCK and PER2 mRNA in hippocampus and adrenal gland were down-regulated significantly (P<0.05) in RA model rats, while moxibustion up-regulated both of them in hippocampus (P<0.05). These results suggested together that moxibustion can benign regulate circadian rhythm of REV-ERBα in different tissues of RA rats. It was revealed that moxibustion not only recovered the losing diurnal oscillation of REV-ERBα in hippocampus and adrenal gland, but also adjusted the circadian rhythm of REV-ERBα in hypothalamus, hippocampus, and adrenal gland to close the normal circadian pattern.
类风湿关节炎（RA）的关键临床症状及既往研究结果显示出昼夜节律变化，清晨时关节肿胀、僵硬及疼痛更为明显。艾灸能够通过多种途径缓解RA，然而，关于RA病理节律的验证性研究结果尚缺。因此，我们开展此项研究以验证艾灸是否可通过调节核心生物钟基因来调整RA的昼夜节律。基于此前在分子水平发现RA的昼夜节律计时受到干扰，本研究旨在观察艾灸对RA大鼠不同组织中REV-ERBα表达水平及昼夜节律的影响。此外，通过逆转录聚合酶链反应（RT-PCR）评估与RA密切相关的核心生物钟基因的表达水平。将96只Sprague-Dawley（SD）大鼠按1:1:1:1的比例随机分为4组，即正常对照组、RA模型组、早上5 - 7点艾灸组和下午5 - 7点艾灸组。采用RT-PCR检测下丘脑、海马和肾上腺中REV-ERBα、CLOCK、BMAL1和PER2的相对表达量。在RA大鼠中，不同组织中REV-ERBα mRNA的表达水平上调，艾灸可使其在不同程度上潜在上调。在未治疗的RA大鼠中，海马和肾上腺中REV-ERBα mRNA的昼夜节律均消失（P>0.05），而艾灸能够使其恢复（P<0.05）。RA模型大鼠海马和肾上腺中CLOCK和PER2 mRNA的表达水平显著下调（P<0.05）；而艾灸使其在海马中的表达上调（P<0.05）。这些结果共同表明，艾灸可良性调节RA大鼠不同组织中REV-ERBα的昼夜节律。研究表明，艾灸不仅恢复了海马和肾上腺中REV-ERBα丧失的昼夜振荡，还调整了下丘脑、海马和肾上腺中REV-ERBα的昼夜节律，使其接近正常昼夜模式。
