| 黄柳云,周海丽,詹魁骏,刘喜德.温化蠲痹方对胶原诱导性关节炎小鼠滑膜增殖、凋亡及Hotair/Wnt1/β-catenin通路的影响[J].中国中西医结合杂志,2023,43(3):309-314 |
| 温化蠲痹方对胶原诱导性关节炎小鼠滑膜增殖、凋亡及Hotair/Wnt1/β-catenin通路的影响 |
| Effects of Wenhua Juanbi Decoction on Proliferation， Apoptosis and Hotair/Wnt1/β-catenin Pathway in the Synovial Membrane of Collagen-Induced Arthritis Mice |
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| DOI：10.7661/j.cjim.20220719.306 |
| 中文关键词: 温化蠲痹方 类风湿关节炎 胶原诱导性关节炎 Hotair/Wnt1/β-catenin信号通路 |
| 英文关键词:Wenhua Juanbi Decoction rheumatoid arthritis collagen-induced arthritis Hotair/Wnt1/β-catenin signaling pathway |
| 基金项目:浙江省自然科学基金项目（No.LY21H270001）；杭州市卫生科技计划重大项目（No.Z20200026） |
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| 中文摘要: |
| 目的 以胶原诱导性关节炎（CIA）小鼠为实验对象进行体内实验，从Hotair/Wnt1/β-连环蛋白（β-catenin）信号通路的角度，分析温化蠲痹方（WHJBD）对类风湿关节炎（RA)的干预机制。方法 选用雄性DBA/1J小鼠建立CIA模型，将造模成功小鼠按随机数字表法分为4组：模型组（CIA组）、温化蠲痹方组（WHJBD组）、甲氨蝶呤组（MTX组）、WHJBD+MTX组，每组6只。并随机选取同一批次的正常小鼠6只作为正常组。WHJBD组每天予WHJBD灌胃（25.78 g/kg）、MTX组每周予MTX灌胃（1.288 mg/kg）、WHJBD+MTX组同时予WHJBD及MTX灌胃，正常组及CIA组予同等剂量生理盐水灌胃，均持续4周。采用游标卡尺测量足掌和踝关节的肿胀程度并计算关节肿胀度，实时荧光定量PCR法检测各组Hotair、Wnt抑制因子1（WIF1）、C-myc、G1/S - 特异性周期蛋白（CyclinD1）表达水平，免疫组化法检测Wnt1、β-catenin、肿瘤增殖抗原抗体（Ki-67）、Caspase3、B 淋巴细胞瘤-2（Bcl2）、Bcl2相关X蛋白（Bax）蛋白表达水平。结果 与CIA组比较，WHJBD、MTX、WHJBD+MTX组干预后小鼠关节肿胀度随着干预时间的增加逐渐降低；增殖相关蛋白Ki-67表达减少，凋亡相关蛋白Caspasse3、Bax表达增加，Bcl2表达减少；Hotair、WIF1基因表达增加，C-myc、CyclinD1基因表达减少；通路蛋白Wnt1、β-catenin表达减少；差异均有统计学意义（P<0.05）。结论 WHJBD具有减轻CIA小鼠足掌和踝关节肿胀度和关节炎程度，调节Hotair/Wnt1/β-catenin通路和滑膜组织的增殖、凋亡，发挥治疗CIA小鼠作用。 |
| 英文摘要: |
| Objective Taking collagen-induced arthritis （CIA） mice as experimental subjects， in vivo experiments were conducted to analyze the therapeutic mechanism of Wenhua Juanbi Decoction （WHJBD） in preventing and treating rheumatoid arthritis（RA） from the perspective of Hotair/Wnt1/β-catenin signaling pathway. Methods Male DBA/1J mice were selected and the CIA mouse model was established， and successfully modeled mice were divided into 4 groups： model group （CIA group）， WHJBD group， methotrexate group （MTX group），WHJBD+MTX group，6 in each group. And 6 normal mice from the same batch were randomly selected as the normal group. Mice in WHJBD group were given WHJBD per day by gastrogavage（25.78 g/kg）. Mice in the MTX group were given MTX per week by gastrogavage （1.288 mg/kg）. Those in the WHJBD+MTX group were given with both WHJBD and MTX by gastrogavage. Those in the normal and model groups were given the same dose of normal saline by gastrogavage. All medication lasted for 4 successive weeks. The swelling of the foot and ankle joints were measured by vernier calipers. Real-time qPCR was used to detect the expression levels of Hotair， Wnt inhibitory factor 1 （ WIF1）， C-myc， and G1/S - specific cyclin - D1 （CyclinD1） in each group， and immunohistochemistry was used to detect the expression levels of Wnt1， β-catenin， antigen identified by monoclonal antibody Ki67（ Ki-67）， Caspase3， B-cell lymphoma-2（Bcl2） and Bcl2 associated X protein（ Bax） proteins. Results Compared with the CIA group， the swelling of mice in the WHJBD， MTX and WHJBD+MTX groups after treatment decreased gradually as intervention time went on. The expression of proliferation-related protein Ki-67 decreased， the expressions of apoptosis-related proteins Caspase3 and Bax increased， and the expression of Bcl2 decreased （P<0.05）. The expressions of Hotair and WIF1 genes increased. The expressions of C-myc and CyclinD1 genes decreased （P<0.05）. The expression of pathway proteins Wnt1 and β-catenin decreased （P<0.05）. Conclusion WHJBD reduced the degree of swelling and arthritis in the paw and ankle joints of CIA mice， regulated the proliferation and apoptosis of Hotair/Wnt1/β-catenin pathway and synovial tissue， thus playing a therapeutic role in CIA mice. |
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