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加味芍甘附子汤对早期生长反应蛋白2调控胶原诱导性关节炎老年大鼠炎症的影响 被引量:1

Effect of modified Shaogan Fuzi decoction on the inflammation in old rats with collagen-induced by early growth reactive protein 2 regulation
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摘要 目的观察加味芍甘附子汤通过早期生长反应蛋白2(early growth responseprotein 2,Egr2)调控胶原诱导性关节炎(collagen induced arthritis,CIA)老年大鼠炎症反应的作用。方法选用清洁Wistar大鼠96只,其中青年鼠与老年鼠各48只。随机将老年鼠与青年鼠分为正常组、模型组、对照组、中药组,每组各12只。正常组常规饲养,其余3组复制CIA大鼠模型后分别给予生理盐水、雷公藤多苷片及加味芍甘附子汤灌胃4周,每周评价各组大鼠关节炎症指数(arthritis index,AI);4周后苏木精—伊红染色(hematoxylin-eosin,HE)检测踝关节组织的病理变化;RT-qPCR检测大鼠膝关节滑膜Egr2 mRNA表达;Elisa法测定外周血白细胞介素-17A(interleukin 17A,IL-17A)、γ干扰素(interferonγ,IFN-γ)、白细胞介素-6(interleukin 6,IL-6)表达。结果(1)各组大鼠AI评价:给药期间,青年鼠和老年鼠的AI评分均降低,且在四周后中药组的评分最低(P<0.05),而老年鼠和青年鼠的组间比较差异无统计学意义。(2)HE检测大鼠踝关节组织的病理变化:四周后取材,踝关节病理学提示除正常组外,各组均有滑膜组织增生及炎性细胞浸润,老年鼠模型组的炎性细胞浸润明显重于青年鼠模型组,且中药组均明显轻于模型组。(3)RT-qPCR检测大鼠膝关节滑膜Egr2 mRNA表达:对照组和中药组的大鼠膝关节滑膜Egr2 mRNA表达高于模型组(P<0.05),老年鼠模型组的大鼠膝关节滑膜Egr2 mRNA表达明显低于青年鼠模型组(P<0.05),而老年鼠中药组的表达明显高于青年中药组(P<0.05)。(4)Elisa法测定大鼠外周血IFN-γ、IL-17A、IL-6的表达:老年鼠中药组IL-17A的表达低于模型组与对照组(P<0.05);青年鼠中药组IL-17A的表达低于模型组(P<0.05),但高于对照组(P<0.05)。老年鼠模型组及对照组外周血IL-17A的表达均高于青年鼠相应组别(P<0.05),而老年鼠中药组的表达低于青年鼠中药组(P<0.05)。结论(1)老年CIA大鼠Egr2的低表达为参与炎症过程可能因素之一;(2)加味芍甘附子汤可能通过对Egr2的调控改善CIA老年鼠关节滑膜炎症,还需进一步开展机制研究。 Objective To observe the effects of modified Shaogan Fuzi decoction on the inflammatory response in old rats with CIA by Egr2.Methods 96 clean Wistar rats were selected,including 48 younger rats and 48 elderly rats.The elderly rats and younger rats were randomly divided into the normal group,the model group,the control group and the traditional Chinese medicine group.The normal group was routinely fed,and the other three groups were given normal saline,Tripterygium Glycosides tablets and modified Shaogan Fuzi decoction modified Shaogan Fuzi decoction for 4 weeks after the replication of CIA rat model.The AI of rat was evaluated weekly in each group.After 4 weeks,HE staining was used to detect the pathological changes of ankle tissue.The levels of IFN-γ、IL-17A、IL-6 were detected in peripheral blood by Elisa,and Egr2 mRNA expression in synovium of knee joint of rats was detected by RT-qPCR.Results The AI scores of young and old rats were decreased during the administration period,and the score of Chinese medicine group was the lowest after four weeks(P<0.05),but there was no significant difference between the old and young groups.Four weeks later,he staining of ankle joint pathology showed that except the normal group,synovial tissue proliferation and inflammatory cell infiltration were found in all groups.The inflammatory cell infiltration in the old mouse model group was significantly heavier than that in the young mouse model group,and the Chinese medicine group was significantly lighter than that in the model group.RT-qPCR method was used to determine the expression of Egr2 mRNA in the synovium of rat knee joint:the expression in the control group and the traditional Chinese medicine group was higher than that in the model group(P<0.05),the expression in the old model group was significantly lower than that in the young model group(P<0.05),and the expression in the old traditional Chinese medicine group was significantly higher than that in the young traditional Chinese medicine group(P<0.05).The expression of IFN-γ,IL-17A and IL-6 in peripheral blood of rats was determined by ELISA method:the expression of inflammatory cytokines in old and young rats showed that the expression of traditional Chinese medicine group and control group was lower than that in model group,while the expression of IL-17A in old traditional Chinese medicine group was lower than that in model group(P<0.05),and lower than that in control group(P<0.05).The expression of IL-17A in young rats was also lower than that in traditional Chinese medicine group(P<0.05),but higher than that in the control group,with statistical significance.The expression of IL-17A in peripheral blood of old rats model group and control group was higher than that in young rats(P<0.05),while the expression of IL-17A in the old rats Chinese medicine group was lower than that in the young rats Chinese medicine group(P<0.05).Conclusion The low expression of Egr2 in elderly patients with rheumatoid arthritis may be associated with more acute onset,more severe clinical symptoms of joint swelling and poor drug resistance;The modified Shaogan Fuzi decoction had more advantages in regulating inflammation of EGR2 in elderly rats of CIA than in younger rats of CIA.
作者 钱唐亮 张玉婷 侯秀娟 刘小平 郑炜 康天伦 王海瑜 QIAN Tangliang;ZHANG Yuting;HOU Xiujuan;LIU Xiaoping;ZHENG Wei;KANG Tianlun;WANG Haiyu(Graduate School of Beijing University of Chinese Medicine,Beijing 100029,China)
出处 《环球中医药》 CAS 2022年第3期370-377,共8页 Global Traditional Chinese Medicine
基金 国家自然科学基金(81774275)。
关键词 加味芍甘附子汤 老年类风湿关节炎 早期生长蛋白2 胶原诱导性关节炎 老年大鼠 Modified Shaogan Fuzi decoction Elderly onset rheumatoid arthritis Early growth response protein 2 Collagen induced arthritis Old rats
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