右归饮对骨性关节炎模型大鼠膝关节滑膜细胞自噬和凋亡相关蛋白及血清炎症因子的影响  

Yougui Drink Affects Osteoarthritis Rats Via Inducing the Expression of Autophgy and Apoptosis Related Proeins and Serum Inflammation Factors

作　　者：黄杨[1] 江浪清[1] 孔劲松[1] 郑鑫[1] 宫小康[1] 阮建伟[1] 王海宝[1] HUANG Yang;JIANG Langqing;KONG Jinsong;ZHENG Xin;GONG Xiaokang;RUAN Jianwei;WANG Haibao(Department of orthopedics,Taizhou Municipal Hospital,Taizhou（31800）China)

机构地区：[1]浙江省台州市立医院骨科,台州318000

出　　处：《浙江中西医结合杂志》2018年第9期739-742,817,共5页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：浙江省中医药科技计划项目(No.2016ZA204);浙江省公益技术应用研究项目(实验动物)(No.2017C37109);浙江省实验动物科技计划项目(No.2018C37107)

摘　　要：目的观察右归饮对骨关节炎(OA)模型大鼠膝关节滑膜细胞自噬和凋亡相关蛋白及血清炎症因子的影响。方法将30只SD大鼠随机分为右归饮组、模型组和空白对照组,各10只。右归饮组、模型组大鼠采用Hulth法进行膝骨关节炎造模。4周后三组大鼠分别予右归饮(每毫升含生药0.69g,剂量10g/体质量)、生理盐水进行灌胃治疗。造模后第8周处死所有大鼠,采集大鼠血清、膝关节及膝关节滑膜,采用ELISA、免疫印迹、HE染色及Mankin’s评分法检测,观察自噬蛋白Beclin-1、m TOR和凋亡相关蛋白Caspase-3、Bax表达,血清白细胞介素1β(IL-1β)、肿瘤坏死因子(TNF-α)、诱导型一氧化氮合酶(i NOS)水平,以及关节软骨损伤情况。结果右归饮组大鼠血清IL-1β、TNF-α和i Nos水平显著低于模型组[IL-1β:(20.27±1.41)pg/m L比(26.76±1.93)pg/m L,P<0.01;TNF-α:(162.63±10.76)pg/m L比(221.34±19.38)pg/m L,P<0.01;i Nos:(7.32±0.48)U/m L比(10.13±0.96)U/m L,P<0.01];右归饮组自噬蛋白表达量高于模型组[Beclin-1:(0.94±0.11)比(0.54±0.08),P<0.01;m TOR:(0.61±0.18)比(0.16±0.09),P<0.01];凋亡蛋白表达量低于模型组[Caspase-3:(0.25±0.07)比(0.60±0.12),P<0.01;Bax:(0.48±0.14)比(0.79±0.11),P<0.01];模型组大鼠膝关节软骨细胞排列紊乱,软骨面大面积缺损,潮线大量破坏;右归饮组大鼠膝关节软骨表面毛糙,部分潮线破坏,软骨细胞排列尚均匀。右归饮组大鼠Mankin’s评分显著低于模型组[(5.20±0.84)分比(8.20±0.84)分,P<0.01]。结论右归饮可能通过上调自噬蛋白表达,下调凋亡蛋白表达,降低血清炎性因子含量,缓解OA模型大鼠膝关节炎性浸润,减轻软骨细胞的破坏。Objective To investigate the effect of Yougui Drink on the treatment of osteoarthritis（OA） rats via inducing the expression of autophagy and apoptosis related proeins and serum inflammation factors. Methods 30 male SD rats were allocated to sham group, OA group and treatment group randomly. OA rat model was constructed by Hulth method. Four weeks later, rats from 3 groups were intragastric administrated by equivalent Yougui Drink and saline, respectively. Eight weeks later, all the 30 rats were sacrificed, blood, knee joints and synovial membranes were collected. The levels of autophagy related proeins Beclin-1 and m TOR, apoptosis related proeins Caspase-3 and Bax, inflammation factors including IL-1β, TNF-α and i NOS were quantified by ELISA, immunoblotting and H-E staining. The severity of articular cartilage damage was evaluated by Mankin＇s scoring. Results In the Yougui Drink treated rats, levels of IL-1β（20.27±1.41 vs 26.76±1.93 pg/m L）, TNF-α（162.63±10.76 vs221.34±19.38 pg/m L） and i NOS（7.32±0.48 vs 10.13±0.96 U/m L） reduced significantly with P〈0.01; expression of the autophagy related proteins increased, and the apoptosis related proeins decreased significantly with P〈0.01, Beclin-1：（0.94±0.11） vs（0.54±0.08）, m TOR：（0.61±0.18） vs（0.16±0.09）, Caspase-3：（0.25±0.07） vs（0.60±0.12）, Bax：（0.48±0.14） vs（0.79±0.11）, compared with those of model rats. Histopathological examination showed deranged chondrocytes in knee joints, large articular cartilage defects, many interrupted tidal lines in the model rats, revealing severely damaged, compared with those of the treatment group. Mankin scores of the Yougui Drink treated rats were lower（8.20±0.84） than those of the model rats（5.20±0.84） significantly（P〈0.01）. Conclusion Yougui Drink could attenuate inflammatory infiltration and damage of articular cartilage in osteoarthritis rat via inducing autophagy up-regulation, apoptosis down-regulation, and inhibiting the inflammat

关 键 词：大鼠 骨性关节炎 自噬 凋亡 炎症 右归饮 

分 类 号：R285.5[医药卫生—中药学] R-332[医药卫生—中医学]