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林也,廖菁,戴宗顺,张婷,胡胜涛,张二兵,易欧阳,张逢,王莘智,李鑫,蔡雄.风寒湿外邪作用于EPO影响痹证（类风湿关节炎）的发生[J].湖南中医药大学学报,2021,41(3):345-349[点击复制] |
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| 风寒湿外邪作用于EPO影响痹证（类风湿关节炎）的发生 |
| 林也,廖菁,戴宗顺,张婷,胡胜涛,张二兵,易欧阳,张逢,王莘智,李鑫,蔡雄 |
| (湖南中医药大学中药粉体与创新药物省部共建国家重点实验室(培育基地), 湖南 长沙 410208;湖南中医药大学中医诊断学湖南省重点实验室, 湖南 长沙 410208;湖南中医药大学第一附属医院风湿免疫科, 湖南 长沙 410007) |
| 摘要: |
| 目的 明确风寒湿外邪作用于促红细胞生成素（EPO）影响痹证（类风湿关节炎）的发生机制。方法 （1）30只雄性SD大鼠随机分为正常对照组、单纯完全弗氏佐剂（CFA）组、风寒湿+CFA组，每组10只大鼠。单纯CFA组大鼠正常饲养14 d，风寒湿+CFA组大鼠风寒湿刺激14 d，再尾根部皮下注射0.1 mL含100 μg热灭活结核杆菌（Mtb）的CFA诱导佐剂性关节炎（AIA）。分别于实验前、风寒湿刺激14 d及CFA免疫后7 d尾动脉取血，采用ELISA法检测血清EPO含量。（2）30只雄性SD大鼠随机分为单纯CFA组、风寒湿+CFA组、风寒湿+CFA+rhEPO组，每组10只大鼠。单纯CFA组正常饲养14 d，风寒湿+CFA组、风寒湿+CFA+rhEPO组，风寒湿刺激14 d，再尾根部皮下注射0.1 mL含100 μg Mtb的CFA诱导AIA。风寒湿+CFA+rhEPO组自风寒湿刺激第1天起，尾静脉注射重组人促红细胞生成素（rhEPO）。观察大鼠的发病时间和发病率；CFA免疫后8 d，腹主动脉取血，采用ELISA法检测血清TNF-α含量。（3）选取RA寒湿痹阻证伴慢性疾病性贫血患者、RA寒湿痹阻证伴缺铁性贫血患者、RA寒湿痹阻证无贫血患者及健康志愿者各20例，取静脉血，采用ELISA法检测血清EPO含量。结果 （1）风寒湿刺激14 d及CFA免疫后7 d，风寒湿+CFA组大鼠血清EPO水平明显低于正常对照组大鼠和单纯CFA组大鼠（P<0.05）。（2）风寒湿+CFA+rhEPO组大鼠AIA发病时间和发病率与单纯CFA组相比无显著差异；风寒湿+CFA+rhEPO组大鼠血清TNF-α含量显著低于风寒湿+CFA组大鼠（P<0.01），而与单纯CFA组相比无显著性差异。（3）RA寒湿痹阻证伴慢性病贫血患者血清EPO含量显著高于健康志愿者（P<0.01），但却显著低于RA寒湿痹阻证伴缺铁性贫血组（P<0.05）。结论 风寒湿外邪通过抑制EPO表达而影响痹证（类风湿关节炎）的发生。 |
| 关键词: 痹证 类风湿关节炎 风寒湿外邪 佐剂性关节炎 EPO |
| DOI：10.3969/j.issn.1674-070X.2021.03.004 |
| 投稿时间：2020-10-27 |
| 基金项目:国家自然科学基金项目（81803993）；湖南省“芙蓉学者奖励计划”资助项目（湘教通〔2020〕58号）；湖南省自然科学基金项目（2018JJ2293，2019JJ40225）；湖南省121创新人才培养工程资助项目（湘人社函〔2019〕192号）；湖南省高校创新平台开放基金项目（17K069）；湖南省中医药科研计划项目（201909）；湖南中医药大学中医学国内一流建设学科项目（校行科字〔2018〕3号）。 |
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| Exogenous Wind-cold-damp on EPO Affects the Occurrence of Bi Syndrome (Rheumatoid Arthritis) |
| LIN Ye,LIAO Jin,DAI Zongshun,ZHANG Ting,HU Shengtao,ZHANG Erbing,YI Ouyang,ZHANG Feng,WANG Shenzhi,LI Xin,CAI Xiong |
| (Institute of Innovation and Applied Research in Chinese Medicine, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Hunan Provincial Key Laboratory of Diagnostic and Therapeutic Research in Chinese Medicine, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Department of Rheumatology, The First Affiliated Hospital, Hunan University of Chinese Medicine, Changsha, Hunan 410007, China) |
| Abstract: |
| Objective To clarify the mechanism of the effect of exogenous wind-cold-damp on erythropoietin (EPO) on Bi syndrome (rheumatoid arthritis, RA). Methods (1) 30 male SD rats were randomly assigned into control group, alone complete Freund's adjuvant (CFA) group and wind-cold-damp+CFA group, 10 rats in each group. Alone CFA group was routinely fed for 14 days. The wind-cold-damp+CFA group was received wind-cold-damp stimulation for 14 days, 0.1 mL CFA comtaining 100 μg Mycobactrium tuberculosis (Mtb) was subcutaneously injected into the base of the tails root to induce adjuvant-induced arthritis (AIA). Blood samples were collected from tails artery before the experiment, 14 days after wind-cold-damp stimulation, and 7 days after the CFA immunization. EPO expression was detected by ELISA. (2) 30 male SD rats were randomly assigned into alone CFA group, wind-cold-damp+CFA group and wind-cold-damp+CFA+recombinant human erythropoietin (rhEPO) group, 10 rats in each group. The alone CFA group was routinely fed for 14 days. The wind-cold-damp+CFA group and wind-cold-damp+CFA+rhEPO group were received wind-cold-damp stimulation for 14 days, 0.1 mL CFA comtaining 100 μg Mtb was subcutaneously injected into the base of the tails root to induce AIA. The wind-cold-damp+CFA+rhEPO group was injected into rhEPO through tail vein from the first day of wind-cold-damp stimulation. The morbidity was daily observed, and the incidence was calculated. On day 8 after CFA immunization, blood samples were collected from abdominal aorta, and serum TNF-level was measured by ELISA. (3) Each twenty patients of the RA cold-dampness syndrome combine with chronic anaemia, the RA cold-damp syndrome combine with iron-deficiency anaemia, the RA cold-damp syndrome without anemia and healthy volunteers were selected, and collected those venous blood. The content of EPO in serum was detected by ELISA. Results (1) The EPO expression in wind-cold-damp+CFA group that in 14 days after wind-cold-damp stimulation, and 7 days after the CFA immunization were obviously below than that in control group and alone CFA group (P<0.05). (2) The morbidity and incidence of AIA were no significant differences between the wind-cold-damp+CFA+rhEPO group and alone CFA group. TNF-level in wind-cold-damp+CFA+rhEPO group was significantly lower than that in wind-cold-damp+CFA group (P<0.01), but had no significant difference from that in alone CFA group. (3) The serum EPO in RA cold-damp syndrome combine with chronic anaemia was significantly higher than that of healthy volunteers (P<0.01), it was significantly lower than that of RA cold dampness syndrome combine with iron deficiency anemia group (P<0.05). Conclusion Exogenous wind-cold-damp stimulation promotes the onset and incidence of Bi syndrome (RA) through inhibiting erythropoietin expression. |
| Key words: Bi syndrome rheumatoid arthritis exogenous wind-cold-damp adjuvant-induced arthritis EPO |
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