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苗药八角枫水提液对CIA模型大鼠血清IL-1β、TNF-α水平及滑膜OPG/RANKL/RANK系统的影响
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| 篇名： | 苗药八角枫水提液对CIA模型大鼠血清IL-1β、TNF-α水平及滑膜OPG/RANKL/RANK系统的影响 |
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| 摘要： | 目的：研究八角枫水提液对Ⅱ型胶原诱发关节炎（CIA）模型大鼠血清白细胞介素1β（IL-1β）、肿瘤坏死因子α（TNF-α）水平以及滑膜骨保护素（OPG）/核因子κB受体活化因子配体（RANKL）/核因子κB受体活化因子（RANK）系统的影响，探讨其抗类风湿关节炎（RA）的可能机制。方法：将60只雄性SD大鼠随机分为空白组、模型组、阳性组（雷公藤多苷片，10 mg/kg）和八角枫水提液低、中、高剂量组（5、10、20 g/kg，按生药量计），每组10只。除空白组外，其余各组大鼠均于尾根部注射经弗氏完全佐剂乳化的牛Ⅱ型胶原蛋白以复制CIA模型。造模后第10天，各给药组大鼠均灌胃相应药物，空白组和模型组大鼠均灌胃等体积生理盐水，每天1次，连续2周。记录各组大鼠的一般情况、关节炎指数（AI）和足跖肿胀度；采用酶联免疫吸附测定法检测大鼠血清中IL-1β、TNF-α水平；采用逆转录聚合酶链反应法检测大鼠滑膜组织中OPG、RANKL、RANK mRNA的表达量；采用苏木精-伊红染色法观察大鼠踝关节组织病变情况。结果：八角枫水提液高剂量组有3只大鼠死亡，阳性组和八角枫水提液中剂量组各有1只大鼠造模失败。造模后，除空白组外的其余组大鼠均出现不同程度的足底及踝关节肿胀等症状。与空白组比较，其余组大鼠AI，足跖肿胀度，血清中IL-1β、TNF-α水平，滑膜组织中RANKL、RANK mRNA的表达量均显著升高或增加，OPG mRNA的表达量均显著下降（P＜0.01）。与模型组比较，各给药组大鼠AI（造模后第28天），足跖肿胀度（造模后第21、28天），血清中IL-1β、TNF-α水平，滑膜组织中RANKL、RANK mRNA的表达量均显著下降，OPG mRNA的表达量均显著升高（P＜0.05），且八角枫水提液高剂量组的AI、IL-1β水平、RANKL mRNA的表达量以及八角枫水提液中、高剂量组RANK mRNA的表达量均显著低于阳性组，阳性组和八角枫水提液高剂量组的TNF-α水平均显著低于八角枫水提液低、中剂量组（P＜0.05或P＜0.01）。空白组大鼠踝关节表面光滑，滑膜细胞无增生，且未见炎症细胞浸润；模型组大鼠关节间隙变窄，关节软骨退变，滑膜细胞增生，局部可见骨质破坏及炎症细胞浸润；各给药组大鼠上述症状均有不同程度的改善。结论：八角枫水提液能够减轻CIA模型大鼠的炎症反应、关节软骨退变及骨破坏，其机制可能与下调血清IL-1β、TNF-α水平，调节OPG/RANKL/RANK系统平衡有关。 |
| ABSTRACT： | OBJECTIVE： To study the effects of Alangium chinense water extract on the levels of IL-1β and TNF-α in serum and OPG/RANKL/RANK system in synovium of type Ⅱ collagen-induced arthritis （CIA） model rats， and to investigate potential mechanism of anti-rheumatoid arthritis （RA）. METHODS： Totally 60 male SD rats were randomly divided into blank group， model group， positive group （Tripterysium glycosides tablets， 10 mg/kg）， A. chinense water extract low-dose， medium-dose and high-dose groups （5， 10， 20 g/kg， by crude drug）， with 10 rats in each group. Except for blank group， CIA model was established by giving Bovine type Ⅱ collagen emulsified with Freund’s complete adjuvant via tail. On the tenth day after modeling， medication groups were given the relevant medicine intragastrically， while blank group and model group were given constant volume of normal saline intragastrically， once a day for consecutive 2 weeks. General information， arthritis index （AI） and plantar swelling degree were recorded in each group. The serum levels of IL-1β and TNF-α were determined by ELISA. The mRNA expression of OPG， RANKL and RANK in synovium were determined by RT-PCR. The pathological changes of the ankle joint were observed by HE staining. RESULTS： Three rats in A. chinense water extract high-dose group died， while one in positive group and one in A. chinense water extract medium-dose group failed to establish the model. After modeling， other groups suffered from different degree swelling of plantar and ankle joints. Compared with blank group， AI， plantar swelling degree， serum levels of IL-1β and TNF-α， mRNA expression amount of RANKL and RANK in synovium were increased significantly， while mRNA expression amount of OPG was decreased significantly （P＜0.01）. Compared with model group， AI （28 d after modeling）， plantar swelling degree （21， 28 d after modeling）， serum levels of IL-1β and TNF-α， mRNA expression amount of RANKL and RANK in synovium were decreased significantly， while mRNA expression amount of OPG was increased significantly （P＜0.05）. AI， serum level of IL-1β and mRNA expression amount of RANKL in A. chinense water extract high-dose group as well as mRNA expression amount of RANK in A. chinense water extract medium-dose and high-dose groups were all significantly lower than positive group. Serum levels of TNF-α in positive group and A. chinense water extract high-dose group were significantly lower than A. chinense water extract low-dose and medium-dose groups （P＜0.05 or P＜0.01）. In blank group， the surface of the ankle joint was smooth， the synovial cells did not proliferate， and no inflammatory cell infiltration was observed. In model group， articular space became narrow， articular cartilage degenerated， synovial cells proliferated， local bone destruction and inflammatory cell infiltration were observed. The above symptoms of medication groups were improved to varying degrees. CONCLUSIONS： A. chinense water extract can relieve the arthritis reaction， degeneration of articular cartilage and bone destruction in CIA model rats， the mechanism of which may be related to reduce the serum levels of IL-1β and TNF-α， and regulate OPG/RANKL/RANK system. |
| 期刊： | 2018年第29卷第24期 |
| 作者： | 江勇，梁子聪，陈其宽，覃建锋，张庆忠，王恒 |
| AUTHORS： | JIANG Yong，LIANG Zicong，CHEN Qikuan，QIN Jianfeng，ZHANG Qingzhong，WANG Heng |
| 关键字： | 八角枫；水提液；类风湿关节炎；Ⅱ型胶原诱发关节炎；炎症因子；骨保护素；核因子κB受体活化因子配体；核因子κB受体活化因子；血清；滑膜组织；大鼠 |
| KEYWORDS： | Alangium chinense； Water extract； Rheumatoid arthritis； Type Ⅱ collagen-induced arthritis； Inflammatory factor； OPG； RANKL； RANK； Serum； Synovium； Rats |
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