Xu Xiao, Wang Miao-Miao, Sun Zhi-Ling, Zhou Dan-Ping, Wang Ling, Wang Fu-Qiang, Xu Zhi-Yang, Ma Qian
School of Nursing, Nanjing University of Chinese Medicine, Nanjing, China.
Analytical and Testing Center, Nanjing Medical University, Nanjing, China.
Acupunct Med. 2016 Jun;34(3):184-93. doi: 10.1136/acupmed-2015-010909. Epub 2015 Nov 5.
To examine the possible impact of moxibustion on the serum proteome of the collagen-induced arthritis (CIA) rat model.
Thirty-six male Sprague-Dawley rats were included in this experiment. The CIA animal model was prepared by injection of type II bovine collagen in Freund's adjuvant on the first and seventh day. The 36 rats were randomly divided into two groups: the untreated CIA group (control), and the CIA plus treatment with moxibustion (CIA+moxi) group. Moxibustion was administered daily at ST36 and BL23 for 7, 14 or 21 days (n=12 rats each). Arthritis score was used to assess the severity of arthritis. At the end of each 7 day treatment, blood samples from the control group and the CIA+moxi group were collected. After removal of high abundance proteins from serum samples, two-dimensional gel combined with matrix-assisted laser desorption ionisation time-of-flight MS/MS (MALDI-TOF-MS/MS) techniques were performed to examine serum protein expression patterns of the CIA rat model with and without moxibustion treatment. In addition, the relevant proteins were further analysed with the use of bioinformatics analysis.
Moxibustion significantly decreased arthritis severity in the rats in the CIA+moxi group, when compared with the rats in the CIA group 35 days after the first immunisation (p=0.001). Seventeen protein spots which changed >1.33 or <0.77 at p<0.05 using Bonferonni correction for multiple testing were found to be common to all three comparisons, and these proteins were used for classification of functions using the Gene Ontology method. Consequently, with the use of the Ingenuity Pathway Analysis, the top canonical pathways and a predicted proteomic network related to the moxibustion effect of CIA were established.
Using the proteomics technique, we have identified novel candidate proteins that may be involved in the mechanisms of action underlying the beneficial effects of moxibustion in rats with CIA. Our findings suggest that immune responses and metabolic processes may be involved in mediating the effects of moxibustion. Moreover, periodxiredoxin I (PRDX1) and inositol 1,4,5-triphosphate receptor (IP3R) may be potential targets.
研究艾灸对胶原诱导性关节炎（CIA）大鼠模型血清蛋白质组的可能影响。
本实验纳入36只雄性Sprague-Dawley大鼠。在第1天和第7天通过在弗氏佐剂中注射II型牛胶原蛋白制备CIA动物模型。将36只大鼠随机分为两组：未治疗的CIA组（对照组）和CIA加艾灸治疗组（CIA+moxi组）。每天在足三里（ST36）和肾俞（BL23）进行艾灸，持续7、14或21天（每组n = 12只大鼠）。关节炎评分用于评估关节炎的严重程度。在每次7天治疗结束时，收集对照组和CIA+moxi组的血样。从血清样本中去除高丰度蛋白质后，采用二维凝胶结合基质辅助激光解吸电离飞行时间串联质谱（MALDI-TOF-MS/MS）技术检测艾灸治疗和未治疗的CIA大鼠模型的血清蛋白质表达模式。此外，利用生物信息学分析对相关蛋白质进行进一步分析。
与首次免疫后35天的CIA组大鼠相比，艾灸显著降低了CIA+moxi组大鼠的关节炎严重程度（p = 0.001）。在所有三个比较中均发现17个蛋白质点在使用Bonferonni校正进行多重检验时p<0.05且变化>1.33或<0.77，这些蛋白质用于使用基因本体论方法进行功能分类。因此，利用 Ingenuity 通路分析，建立了与CIA艾灸效应相关的顶级经典通路和预测的蛋白质组网络。
使用蛋白质组学技术，我们鉴定出了可能参与艾灸对CIA大鼠有益作用机制的新候选蛋白质。我们的研究结果表明，免疫反应和代谢过程可能参与介导艾灸的作用。此外，过氧化物氧还蛋白I（PRDX1）和肌醇1,4,5-三磷酸受体（IP3R）可能是潜在靶点。
