摘要
背景:前期研究表明独活寄生汤可有效抑制软骨细胞凋亡,但其具体机制尚不明确。目的:观察独活寄生汤对膝骨关节炎大鼠软骨PERK/Bip信号通路关键调节因子PERK、Bip、eI F-2α、ATF-4、GADD153、Caspase-9、Caspase-3的影响,探讨独活寄生汤防治骨关节炎的作用机制。方法:80只SD大鼠适应性喂养1周后,随机分为正常组(20只)和造模组(60只)。造模组应用膝关节注射木瓜蛋白酶法制备膝骨关节炎模型。造模成功后随机分为3组,即模型组(n=20)、治疗组(n=20)和阳性对照组(n=20)。正常组和模型组给予生理盐水;治疗组给予独活寄生汤,按照9.3 g/(kg·d)的药量进行灌胃;阳性对照组给予盐酸氨基葡萄糖胶囊0.15 g/(kg·d)。2周为1个疗程,共4个疗程。分别于2,4个疗程后切取右侧胫骨平台,免疫组化法和RT-PCR法检测PERK、Bip、e IF-2α、ATF-4、GADD153、Caspase-9、Caspase-3蛋白和mRNA表达情况。结果与结论:(1)RT-PCR结果显示,独活寄生汤和盐酸氨基葡萄糖均能抑制PERK、Bip、e IF-2α、ATF-4、GADD153、Caspase-9、Caspase-3 mRNA表达(P<0.05或0.01),干预时间越长,效果越明显,但2组间无显著差异(P>0.05);(2)免疫组化结果与RT-PCR结果趋势一致,即独活寄生汤和盐酸氨基葡萄糖均能抑制PERK、Bip、e IF-2α、ATF-4、GADD153、Caspase-9、Caspase-3蛋白表达(P<0.05或0.01),干预时间越长,效果越明显,但2组间无显著性差异(P>0.05);(3)结果提示,独活寄生汤可能是通过调控PERK/Bip信号通路,进而抑制因内质网应激反应引起的软骨细胞凋亡。
BACKGROUND： Preliminary study has shown that Duhuo Jisheng Decoction（DHJSD） is an effective prescription for osteoarthritis. But the underling mechanism remains unclear. OBJECTIVE： To observe the effect of DHJSD on the key regulating factors PERK, Bip, eIF-2α, ATF-4, GADD153, Caspase-9 and Caspase-3 in PERK/Bip signaling pathway in cartilage of rats with knee osteoarthritis, so as to explore its treatment mechanism. METHODS： After 1 week of acclimation, 80 Sprague-Dawley rats were randomly divided into normal group（n=20） and model rats（n=60）. Osteoarthritis was induced by injecting 0.2 m L of 4% papain solution in both knees. The rat models were randomly divided into three groups, including model, treatment and positive control groups（n=20 per group）. The normal and model groups received treatment of normal saline. The treatment and positive control groups were given DHJSD（9.3 g/（kg·d）） and glucosamine hydrochloride capsule 0.15 g/（kg·d） via gavage, respectively, for four courses（two weeks a course）. After every two courses, the animals were sacrificed and the right tibial plateau was obtained. The mRNA and protein levels of PERK, Bip, eIF-2α, ATF-4, GADD153, Caspase-9 and Caspase-3 were measured by RT-PCR and immunohistochemistry, respectively. RESULTS AND CONCLUSION： RT-PCR results showed that DHJSD and glucosamine hydrochloride capsule could time-dependently inhibit the mRNA expression of PERK, Bip, eIF-2α, ATF-4, GADD153, Caspase-9 and Caspase-3（P〈0.05 or P〈0.01）. There was no significant difference between treatment and positive control groups（P〈0.05）. Immunohistochemistry results found that DHJSD and glucosamine hydrochloride capsule could time-dependently inhibit the protein expression of PERK, Bip, eIF-2α, ATF-4, GADD153, Caspase-9 and Caspase-3（P〈0.05 or P〈0.01）. There was no significant difference between treatment and positive control groups（P〈0.05）. These results suggest that DHJSD can inhibit the apoptosis of chondrocytes cause
作者
陈俊
吴广文
许惠凤
郑春松
李西海
邱建清
刘淑如
刘献祥
Chen Jun;Wu Guang-wen;Xu Hui-feng;Zheng Chun-song;Li Xi-hai;Qiu Jian-qing;Liu Shu-ru;Liu Xian-xiang(College of Integrative Medicine;Academy of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,Fujian Province,China;Fujian Provincial Key Laboratory of Integrative Medicine on Geriatrics,Fuzhou 350122,Fujian Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2018年第28期4493-4500,共8页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金资助项目(81573801)
福建省自然科学基金杰青项目(2017J06018)
2017年度福建省中医药科技项目(2017FJZYJC204)
福建省卫生计生科研人才培养项目资助(2017-ZQN-62)~~
