痛风清热方对急性痛风性关节炎大鼠滑膜组织中 TNF-α、IL-1β、IL-18的影响

郭玉星 朱珊莹 康佩芝 刘文龙 易法银 邵先舫 熊辉 刘永利 毛果

〔摘要〕 目的 觀察不同剂量痛风清热方对急性痛风性关节炎模型大鼠的踝关节滑膜组织中TNF-α、IL-1β、IL-18炎性因子表达的影响。方法 选取SD雄性大鼠72只，随机分为6组，正常对照组、模型组、秋水仙碱组及痛风清热方高、中、低剂量组，每组12只。模型组、痛风清热方各剂量组及秋水仙碱组按经典Coderre大鼠动物模型造模，正常对照组在相应部位注射等量生理盐水。痛风清热方高、中、低剂量组分别予以60、30、15 g/kg痛风清热方混悬液灌胃干预，秋水仙碱组予以0.09 g/kg秋水仙碱悬浊液灌胃。于造模前、造模后4、8、12、24、48 h测量踝关节周径，测定受试踝关节肿胀指数;于造模后48 h取受试踝关节滑膜组织;采用HE染色观察踝关节滑膜组织病理变化;采用ELISA法检测组织中TNF-α、IL-1β、IL-18的含量。结果 与正常对照组相比，模型组呈现明显的关节炎病理变化，在光镜下可见不同程度的滑膜上皮脱落、血管增生以及大量炎症细胞浸润;而痛风清热方各剂量组和秋水仙碱组则病理变化较少。与正常对照组比较，模型组、痛风清热方各剂量组、秋水仙碱组造模后4、8、12、24、48 h关节肿胀指数明显升高（P<0.05）;与模型组比较，痛风清热方各剂量组及秋水仙碱组造模后4、8、12、24、48 h关节肿胀指数各有不同程度降低（P<0.05），其中秋水仙碱组肿胀指数降低最明显，其次为痛风清热方高剂量组（P<0.05）。与正常对照组比较，模型组、痛风清热方各剂量组及秋水仙碱组大鼠滑膜组织中TNF-α、IL-1β、IL-18水平均增高（P<0.05）;与模型组比较，痛风清热方各剂量组及秋水仙碱组大鼠滑膜组织中TNF-α、IL-1β、IL-18水平均降低（P<0.05），其中秋水仙碱组TNF-α、IL-1β、IL-18水平降低最显著，其次为痛风清热方高剂量组（P<0.05）。结论 痛风清热方对大鼠痛风性关节炎具有明显消肿作用。其作用可通过降低滑膜组织中TNF-α、IL-1β、IL-18炎性因子的表达，从而改善局部组织炎症作用，而且随着痛风清热方剂量的增加，其作用越强。

〔关键词〕 痛风清热方;痛风性关节炎;踝关节肿胀指数;TNF-α;IL-1β;IL-18

〔中图分类号〕R255.6        〔文献标志码〕A       〔文章编号〕doi：10.3969/j.issn.1674-070X.2021.05.004

The Effect of Tongfeng Qingre Recipe on TNF-α， IL-1β and IL-18 in Synovium

Tissue of Rats with Acute Gouty Arthritis

GUO Yuxing1，2， ZHU Shanying1， KANG Peizhi2， LIU Wenlong1， YI Fayin1*， SHAO Xianfang3*，

XIONG Hui1， LIU Yongli3， MAO Guo2

（1. Hunan University of Chinese Medicine， Changsha， Hunan 410208， China; 2. Affiliated Hospital of Hunan Institute of Traditional Chinese Medicine， Changsha， Hunan 410006， China; 3. Changde First Hospital of Traditional Chinese Medicine， Changde， Hunan 415100， China）

〔Abstract〕 Objective To observe the effects of different doses of Tongfeng Qingre Recipe on the expression of inflammatory factor TNF-α， IL-1β and IL-18 in the synovium of ankle joint of rats suffering acute gouty arthritis. Methods 72 male SD rats were randomly divided into 6 groups， including normal control group， model group， colchicine group and Tongfeng Qingre Recipe high-dose， medium-dose and low-dose groups， with 12 rats in each group. The model group and the high， medium and low dose groups and the colchicine group were established the classic Coderre models of the rats， however， rats from the normal control group was given normal saline in the corresponding parts for same dose of the formula utilized in other groups. The high， medium and low dose Tongfeng Qingre Recipe group received intragastric administration of Tongfeng Qingre Recipe traditional Chinese medicine turbid liquid of different dose of 60， 30， 15 g/kg. The colchicine group received intragastric administrated of 0.09 g/kg colchicine turbid liquid. The ankle circumference and the swelling index of the ankle was measured before and 4， 8， 12， 24 and 48 hours after the establishment of the model. Synovial tissue of the ankles was collected 48 hours after modeling. HE staining was used to observe ankle joint synovial tissue pathological changes， and the content of TNF-α， IL-1β and IL-18 in the synovium of ankle joint were determined by ELISA. Results Compared with the normal control group， the model group showed obvious pathological changes of arthritis. Under the light microscope， the model group showed different degrees of synovium epithelial detachment， vascular proliferation and a large number of inflammatory cell infiltration， while the Tongfeng Qingre Recipe group of different dose and colchicine group showed less pathological changes. Compared with the normal control group， the swelling index of the model group， Tongfeng Qingre Recipe group of different dose and colchicine group increased significantly at 4， 8， 12， 24 and 48 hours after modeling （P<0.05）. Compared with the model group， the joint swelling index in the Tongfeng Qingre Recipe group of different dose and colchicine group decreased at 4， 8， 12， 24 and 48 hours after modeling （P<0.05）， and the swelling index of the colchicine group decreased most obviously， followed by the Tongfeng Qingre Recipe high-dose group （P<0.05）. Compared with the normal control group， the levels of  TNF-α， IL-1β and IL-18 in the synovium of the model group， Tongfeng Qingre Recipe group of different dose and colchicine group were significantly higher （P<0.05）. Compared with the model group， the levels of TNF-α， IL-1β and IL-18 in synovium of rats in the Tongfeng Qingre Recipe group of different dose and colchicine group were lower （P<0.05）. The levels of TNF-α， IL-1β and IL-18 in colchicine group decreased most significantly， followed by the Tongfeng Qingre Recipe high-dose group （P<0.05）. Conclusion Tongfeng Qingre Recipe has obvious detumescence effect on gouty arthritis in rats. Its effect can be achieved by reducing the expression of TNF-α， IL-1β and IL-18 and other inflammatory factors in local tissues， and the effect is stronger with the increase of the dose of Tongfeng Qingre Recipe.

〔Keywords〕 Tongfeng Qingre Recipe; gouty arthritis; swelling index of the ankle; TNF-α; IL-1β; IL-18

急性痛风性关节炎是痛风急性发作的首发症状，其典型的临床表现为急性发作的、剧烈的关节红、肿、热、痛。而随着生活水平的提高、饮食结构的改变，我国急性痛风性关节炎的发病率呈明显上升趋势，发病年龄逐渐年轻化[1]。……