正清风痛宁对骨关节炎动物模型ADAMTs-4、ADAMTs-5表达的影响
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摘要
目的分析正清风痛宁对骨关节炎动物模型膝关节软骨组织聚蛋白多糖酶1(ADAMTs-4)、聚蛋白多糖酶2(ADAMTs-5)蛋白表达的影响,探讨正清风痛宁治疗骨关节炎的作用机制。方法将32只成功建模的新西兰兔随机分为观察组和模型组,另选取16只新西兰兔作为空白组。空白组不接受干预治疗,模型组应用生理盐水灌胃,观察组应用正清风痛宁灌胃。药物干预后4周分离膝关节软组织,观察关节形态、关节液、关节软骨,同时进行HE染色和ADAMTs-4、ADAMTs-5 mRNA检测(RT-PCR法),利用统计方法分析组间数据。结果模型组关节软骨分级和Mankin’s评分高于观察组和空白组,观察组关节软骨分级和Mankin’s评分高于空白组(P<0.05)。模型组ADAMTs-4和ADAMTs-5 mRNA表达显著高于观察组和空白组,观察组ADAMTs-4和ADAMTs-5 mRNA表达显著高于空白组(P<0.05)。结论正清风痛宁可改善骨关节炎动物模型关节病变程度,可能与其抑制ADAMTs-4和ADAMTs-5表达有关。
Objective To analyze the effect of a Chinese medicine Zhengqingfengtongning on the expression of aggrecanase-1(ADAMTs-4) and aggrecanase-2(ADAMTs-5) in cartilage tissue of knee joint in rabbit models of osteoarthritis,and to study the mechanism of action of Zhengqingfengtongning in treatment for osteoarthritis. Methods 32 New Zealand rabbits were randomly divided into the observation group and the model group,and 16 healthy New Zealand rabbits were chosen as the blank group. The blank group did not receive the intervention treatment,the model group received physiological saline in gastric gavage,the observation group was given the Zhengqingfengtongning by gastric gavage. The soft tissue samples of knee joint in the three groups were taken at 4 weeks after drug intervention. The joint morphology,joint fluid and articular cartilage were observed by histopathology. The ADAMTs-4 and ADAMTs-5 mRNA were detected by RT-PCR. The data were then statistically analyzed. Results The grades of articular cartilage and Mankin's score of the model group were significantly higher than those of the observation group and blank group,the gradesof articular cartilage and Mankin's score of the observation group were significantly higher than those of the blank group,and the difference between the two groups was statistically significant(P < 0. 05). The ADAMTs-4 and ADAMTs-5 mRNA expressions of the model group were significantly higher than those of the observation group and blank group. The ADAMTs-4 and ADAMTs-5 mRNA expressions of the observation group were significantly higher than those of the blank group,and the difference between the two groups was statistically significant(P < 0. 05). Conclusions The Zhengqingfengtongning can improve the degree of joint lesions in osteoarthritis animal models,which may be related to inhibiting the expression of ADAMTs-4 and ADAMTs-5.
Objective To analyze the effect of a Chinese medicine Zhengqingfengtongning on the expression of aggrecanase-1(ADAMTs-4) and aggrecanase-2(ADAMTs-5) in cartilage tissue of knee joint in rabbit models of osteoarthritis,and to study the mechanism of action of Zhengqingfengtongning in treatment for osteoarthritis. Methods 32 New Zealand rabbits were randomly divided into the observation group and the model group,and 16 healthy New Zealand rabbits were chosen as the blank group. The blank group did not receive the intervention treatment,the model group received physiological saline in gastric gavage,the observation group was given the Zhengqingfengtongning by gastric gavage. The soft tissue samples of knee joint in the three groups were taken at 4 weeks after drug intervention. The joint morphology,joint fluid and articular cartilage were observed by histopathology. The ADAMTs-4 and ADAMTs-5 mRNA were detected by RT-PCR. The data were then statistically analyzed. Results The grades of articular cartilage and Mankin's score of the model group were significantly higher than those of the observation group and blank group,the gradesof articular cartilage and Mankin's score of the observation group were significantly higher than those of the blank group,and the difference between the two groups was statistically significant(P < 0. 05). The ADAMTs-4 and ADAMTs-5 mRNA expressions of the model group were significantly higher than those of the observation group and blank group. The ADAMTs-4 and ADAMTs-5 mRNA expressions of the observation group were significantly higher than those of the blank group,and the difference between the two groups was statistically significant(P < 0. 05). Conclusions The Zhengqingfengtongning can improve the degree of joint lesions in osteoarthritis animal models,which may be related to inhibiting the expression of ADAMTs-4 and ADAMTs-5.
引文
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[12]张恩水,闫新峰,张明,等.聚蛋白多糖酶和基质金属蛋白酶在骨关节炎不同时期关节液中的表达[J].山东大学学报(医学版),2012,50(7):87-91.
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[14]Glasson SS,Askew R,Sheppard B,et al.Deletion of active ADAMTs5 prevents cartilage degradation in a murine model of osteoarthritis[J].Nature,2005,434(7033):644-648.
[15]van Meurs JB,van Lent PL,Holthuysen AE,et al.Kinetics of aggrecanase-and metalloproteinase-induced neoepitopes in various stages of cartilage destruction in murine arthritis[J].Arthritis Rheum,1999,42(6):1128-1139.
[16]余庆阳,黄巍.膝骨关节炎从痹论治的病因与证候探讨[J].风湿病与关节炎,2015,4(3):40-43.
[17]熊力群,余舒鹏,李文龙,等.清风藤临床应用研究概况[J].亚太传统医药,2017,13(5):63-65.
[18]杨洁,曾慧,郑芳.正清风痛宁与西药对比治疗膝骨性关节炎的荟萃分析[J].中成药,2015,37(6):1197-1201.
[19]乙军,周业庭,徐丹,等.实验性兔膝骨关节炎模型的建立[J].临床和实验医学杂志,2012,11(24):1921-1923.
[20]江捍平,王大平.骨关节炎动物模型[J].中国现代医学杂志,2004,14(6):153-154,156.
[21]陈紫岳,黄怡然,李娜,等.电针和圆利针治疗制动致膝关节功能障碍兔模型的治疗效果[J].世界中医药,2017,12(8):1927-1932.
[22]欧云生,安洪.鼠骨关节炎动物模型建立的现状[J].中国比较医学杂志,2004,14(1):41-44.
[23]钱洁,邢雪松,梁军.大鼠膝关节骨性关节炎动物模型的两种实验方案[J].实验室研究与探索,2014,33(11):23-27.
[2]陈益丹,邱华平,金肖青,等.不同针灸方法对膝骨炎模型细胞因子及基质金属蛋白酶影响的比较研究[J].中国比较医学杂志,2016,26(1):42-45,64.
[3]韦隽.正清风痛宁肠溶片联合双氯芬酸钠治疗类风湿性关节炎效果观察[J].中国医药科学,2013,3(12):197-198.
[4]杜明瑞,冯福海,张静.正清风痛宁片联合非甾体抗炎药治疗骨关节炎疗效及安全性Meta分析[J].中华临床免疫和变态反应杂志,2015,9(2):121-125.
[5]史东,董富强,董金波,等.ADAMTs-4、ADAMTs-5在骨关节炎患者滑膜中的表达和意义[J].现代预防医学,2015,42(6):1061-1064,1092.
[6]高亮,陈默,岳萍,等.温针灸对膝骨关节炎兔膝关节软骨转化生长因子β1和胰岛素生长因子Ⅰ水平的影响[J].针刺研究,2015,40(3):229-232.
[7]黄怡然,金英利,李娜,等.针刀、电针和圆利针对膝骨关节炎模型兔股直肌Bcl-2、Bax、Caspase-3蛋白表达的影响[J].针刺研究,2014,39(2):100-105,123.
[8]周效思,周凯,谭安雄,等.威灵仙对兔膝关节炎结构和功能的影响[J].时珍国医国药,2011,22(10):2454-2456.
[9]Mankin HJ,Dorfman H,Lippiello L,et al.Biochemical and metabolic abnormalities in articular cartilage from osteoarthritic human hips.Ⅱ.Correlation of morphology with biochemical and metabolic data[J].J Bone Joint Surg Am,1971,53(3):523-537.
[10]Durigova M,Nagase H,Mort JS,et al.MMPs are less efficient than ADAMTs5 in cleaving aggrecan core protein[J].Matrix Biol,2011,30(2):145-153.
[11]Verma P,Dalal K,Chopra M.Pharmacophore development and screening for discovery of potential inhibitors of ADAMTS-4 for osteoarthritis therapy[J].J Mol Model,2016,22(8):178.
[12]张恩水,闫新峰,张明,等.聚蛋白多糖酶和基质金属蛋白酶在骨关节炎不同时期关节液中的表达[J].山东大学学报(医学版),2012,50(7):87-91.
[13]Song RH,Tortorella MD,Malfait AM,et al.Aggrecan degradation in human articular cartilage explants is mediated by both ADAMTs-4 and ADAMTs-5[J].Arthritis Rheum,2007,56(2):575-585.
[14]Glasson SS,Askew R,Sheppard B,et al.Deletion of active ADAMTs5 prevents cartilage degradation in a murine model of osteoarthritis[J].Nature,2005,434(7033):644-648.
[15]van Meurs JB,van Lent PL,Holthuysen AE,et al.Kinetics of aggrecanase-and metalloproteinase-induced neoepitopes in various stages of cartilage destruction in murine arthritis[J].Arthritis Rheum,1999,42(6):1128-1139.
[16]余庆阳,黄巍.膝骨关节炎从痹论治的病因与证候探讨[J].风湿病与关节炎,2015,4(3):40-43.
[17]熊力群,余舒鹏,李文龙,等.清风藤临床应用研究概况[J].亚太传统医药,2017,13(5):63-65.
[18]杨洁,曾慧,郑芳.正清风痛宁与西药对比治疗膝骨性关节炎的荟萃分析[J].中成药,2015,37(6):1197-1201.
[19]乙军,周业庭,徐丹,等.实验性兔膝骨关节炎模型的建立[J].临床和实验医学杂志,2012,11(24):1921-1923.
[20]江捍平,王大平.骨关节炎动物模型[J].中国现代医学杂志,2004,14(6):153-154,156.
[21]陈紫岳,黄怡然,李娜,等.电针和圆利针治疗制动致膝关节功能障碍兔模型的治疗效果[J].世界中医药,2017,12(8):1927-1932.
[22]欧云生,安洪.鼠骨关节炎动物模型建立的现状[J].中国比较医学杂志,2004,14(1):41-44.
[23]钱洁,邢雪松,梁军.大鼠膝关节骨性关节炎动物模型的两种实验方案[J].实验室研究与探索,2014,33(11):23-27.