甲泼尼龙醋酸酯对重组白细胞介素-1β介导的颞下颌关节炎症损伤大鼠软骨细胞MMPs表达的影响
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摘要
目的探究甲泼尼龙醋酸酯对重组白细胞介素-1β(rh IL-1β)介导的颞下颌关节炎大鼠软骨细胞中基质金属蛋白酶1(MMP-1)、基质金属蛋白酶-3(MMP-3)、基质金属蛋白酶-9(MMP-9)表达的影响。方法以雄性SD大鼠作为研究对象,采用关节腔注射rh IL-1β法制备颞下颌关节炎大鼠模型,HE染色法观察颞下颌关节变化;对照组不进行任何处理。模型制备后将大鼠分为模型组、甲泼尼龙醋酸酯低剂量组(5 mg/kg)、中剂量组(15 mg/kg)、高剂量组(30 mg/kg),每组10只。对照组、模型组经腹腔静脉注射1 ml生理盐水,甲泼尼龙醋酸酯各剂量组分别经腹腔静脉注射甲泼尼龙醋酸酯。HE染色、番红染色观察各组颞下颌关节组织变化,采用Mankin's评分对颞下颌关节炎程度进行评定。RT-PCR法、Western Blot法、免疫组化法检测软骨组织中MMP-1、MMP-3、MMP-9的表达。结果 HE染色显示模型组大鼠颞下颌关节滑膜出现炎症细胞浸润、血管扩张、细胞黏附。与对照组相比,模型组Mankin's评分升高,与模型组相比,给药物组评分降低,且随着药物浓度的升高,评分逐渐下降(P <0. 05)。与对照组相比,模型组MMP-1、MMP-3、MMP-9 mRNA、蛋白表达量均升高,与模型组相比,给药组MMP-1、MMP-3、MMP-9 mRNA、蛋白表达量均降低,且随着药物浓度的升高,三者表达量均逐渐降低(P <0. 05)。免疫组化结果显示,模型组阳性细胞表达率高于对照组,经药物处理后MMP-1、MMP-3、MMP-9阳性细胞表达率随药物剂量的增加呈剂量性降低(P <0. 05)。结论甲泼尼龙醋酸酯能够缓解rh IL-1β介导的颞下颌关节炎,其作用机制可能与下调软骨细胞中MMP-1、MMP-3、MMP-9的表达有关。
Objective To explore the influence of methylprednisolone acetate on cartilage cells MMPs expression of rats with temporomandibular joint inflammatory injury mediated by recombinant interleukin-1β( rhIL-1β). Methods SD male rats were used as the study objects,and the model of temporomandibular arthritis in rats was built by using joint cavity injection rh IL-1β method,and HE staining method was used to observe the changes of Nie and mandible,and after modeling,those were divided into model group,methylprednisolone acetate low dose treatment group( 5 mg/kg),medium dose treatment group( 15 mg/kg),and high dose treatment group( 30 mg/kg). In the control group and the model group,1 ml of normal saline was intraperitoneally injected,and each dose of methylprednisolone acetate was intraperitoneally injected with methylprednisolone acetate. Changes of the mandibular joint tissues in different groups were observed by HE staining and crocus staining. Degree of temporomandibular arthritis was evaluated by using Mankin's score. MMP-1,MMP-3 and MMP-9 expression in cartilage tissue was detected by using RT-PCR,Western,Blot and immunohistochemistry. Results HE staining showed that inflammation,cell infiltration,vasodilatation and cell adhesion were observed in the synovium of TMJ rats of the model group. Compared with that in the control group,Mankin's score increased in the control group,and compared with that in the model group,the score decreased in the drug group,with the increase of drug concentration,the score decreased gradually( P < 0. 05). Compared with those in the control group,MMP-1,MMP-3,MMP-9 and mRNA protein expression in the model group increased,and compared with those in the model group,MMP-1,MMP-3,MMP-9,mRNA protein expression reduced in the drug group,with the increase of drug concentration,the three expression decreased( P < 0. 05). Immunohistochemical staining showed that the expression rate of positive cells in the model group was higher than that in the control group,and the expression rate of MMP-1,MMP-3 and MMP-9 positive cells decreased with the increase of drug concentration( P < 0. 05). Conclusion Methylprednisolone acetate can alleviate rhIL-1β mediated arthritis of the temporomandibular joint,and its mechanism may be related to down-regulate of MMP-1,MMP-3 and MMP-9 expression in chondrocytes.
Objective To explore the influence of methylprednisolone acetate on cartilage cells MMPs expression of rats with temporomandibular joint inflammatory injury mediated by recombinant interleukin-1β( rhIL-1β). Methods SD male rats were used as the study objects,and the model of temporomandibular arthritis in rats was built by using joint cavity injection rh IL-1β method,and HE staining method was used to observe the changes of Nie and mandible,and after modeling,those were divided into model group,methylprednisolone acetate low dose treatment group( 5 mg/kg),medium dose treatment group( 15 mg/kg),and high dose treatment group( 30 mg/kg). In the control group and the model group,1 ml of normal saline was intraperitoneally injected,and each dose of methylprednisolone acetate was intraperitoneally injected with methylprednisolone acetate. Changes of the mandibular joint tissues in different groups were observed by HE staining and crocus staining. Degree of temporomandibular arthritis was evaluated by using Mankin's score. MMP-1,MMP-3 and MMP-9 expression in cartilage tissue was detected by using RT-PCR,Western,Blot and immunohistochemistry. Results HE staining showed that inflammation,cell infiltration,vasodilatation and cell adhesion were observed in the synovium of TMJ rats of the model group. Compared with that in the control group,Mankin's score increased in the control group,and compared with that in the model group,the score decreased in the drug group,with the increase of drug concentration,the score decreased gradually( P < 0. 05). Compared with those in the control group,MMP-1,MMP-3,MMP-9 and mRNA protein expression in the model group increased,and compared with those in the model group,MMP-1,MMP-3,MMP-9,mRNA protein expression reduced in the drug group,with the increase of drug concentration,the three expression decreased( P < 0. 05). Immunohistochemical staining showed that the expression rate of positive cells in the model group was higher than that in the control group,and the expression rate of MMP-1,MMP-3 and MMP-9 positive cells decreased with the increase of drug concentration( P < 0. 05). Conclusion Methylprednisolone acetate can alleviate rhIL-1β mediated arthritis of the temporomandibular joint,and its mechanism may be related to down-regulate of MMP-1,MMP-3 and MMP-9 expression in chondrocytes.
引文
[1]Turp JC,Schindler H,刘晓东,等.颞下颌关节病的病因问题:咬合因素的流行病学和病因学依据[J].实用口腔医学杂志,2015,31(3):417-424.
[2]雍翔智,唐黎黎,农晓琳.颞下颌关节紊乱综合征的心理因素及其治疗[J].国际口腔医学杂志,2013,40(5):634-637.
[3]白正发,李会晓,庞仲辉,等.骨疏康胶囊联合依托考昔治疗膝骨关节炎的临床疗效及对炎性因子表达的影响[J].临床和实验医学杂志,2016,15(4):362-364.
[4]杨军,娄德全,周振东,等.基质金属蛋白酶和胶原在创伤关节软骨组织中的表达[J].中国组织工程研究,2011,15(20):3636-3640.
[5]甘业华,孟娟红,马绪臣.我国颞下颌关节骨关节炎基因表达调控及雌激素与关节炎性疼痛关系的研究进展[J].中华口腔医学杂志,2012,47(1):26-27.
[6]王燕,王晓荣,宋涛,等.高频超声在评价药物治疗膝骨关节炎病变中的应用价值[J].中国超声医学杂志,2013,29(9):825-828.
[7]康信忠,吴启富,王康惠,等.中西医结合治疗类风湿关节炎疗效及其对糖皮质激素受体的影响[J].中国中西医结合杂志,2010,30(12):1261-1264.
[8]杨文英,张文丽,罗应伟.颞下颌关节骨关节炎动物模型的研究进展[J].国际口腔医学杂志,2015,42(6):677-680.
[9]阴健,廖爽,张伟华,等.关节腔内注射氨基葡萄糖治疗兔颞下颌关节骨关节炎的组织学研究[J].国际口腔医学杂志,2014,39(1):36-39.
[10]满城,蒋练,徐凡.rh IL-1Ra抑制大鼠颞下颌关节骨关节炎软骨破坏的研究[J].实用口腔医学杂志,2017,33(4):442-446.
[11]兰玲莉,张伟娜,蒋扬眉,等.Notch信号通路在颞下颌骨关节炎中的表达[J].口腔疾病防治,2017,25(3):143-152.
[12]Jabao!nska-Trypu!c A,Matejczyk M,Rosochacki S.Matrix metalloproteinases(MMPs),the main extracellular matrix(ECM)enzymes in collagen degradation,as a target for anticancer drugs[J].J Enzyme Inhib Med Chem,2016,23(8):177-183.
[13]Patterson J,Hubbell JA.Enhanced proteolytic degradation of molecularly engineered PEG hydrogels in response to MMP-1 and MMP-2[J].Biomaterials,2010,31(30):7836-7845.
[14]李干,李奇,林荔军,等.IL-1、MMP-1及TIMP-1在兔骨性关节炎模型滑膜、软骨中的表达[J].实用医学杂志,2011,27(15):2721-2723.
[15]王晓倩,李鑫,郭建生,等.金凤颗粒对痛风性关节炎兔关节软骨MMP-1/TIMP-1表达影响[J].中国免疫学杂志,2015,31(6):774-777.
[16]Eba H,Murasawa Y,Iohara K,et al.The anti-inflammatory effects of matrix metalloproteinase-3 on irreversible pulpitis of mature erupted teeth[J].Plos One,2012,7(12):52523-52530.
[17]Valimaki J,Uusitalo H.Matrix metalloproteinases(MMP-1,MMP-2,MMP-3 and MMP-9,and TIMP-1,TIMP-2 and TIMP-3)and markers for vascularization in functioning and non-functioning bleb capsules of glaucoma drainage implants[J].Acta Ophthalmol,2015,93(5):450-456.
[18]贺占坤,沈杰威.MMP-2、MMP-3、MMP-9和TIMP-1评价膝关节骨性关节炎的临床研究[J].重庆医学,2013,42(32):3872-3874.
[19]李国华,刘德群,刘会仁.大鼠骨骼肌缺血后MMP-9的变化与骨骼肌损伤和重塑的关系[J].安徽医科大学学报,2012,47(11):1308-1311.
[20]石伟哲,肖海军,薛锋,等.基质金属蛋白酶9在大鼠跟腱异位骨化模型中的动态变化研究[J].中国修复重建外科杂志,2014,28(9):116-120.
[2]雍翔智,唐黎黎,农晓琳.颞下颌关节紊乱综合征的心理因素及其治疗[J].国际口腔医学杂志,2013,40(5):634-637.
[3]白正发,李会晓,庞仲辉,等.骨疏康胶囊联合依托考昔治疗膝骨关节炎的临床疗效及对炎性因子表达的影响[J].临床和实验医学杂志,2016,15(4):362-364.
[4]杨军,娄德全,周振东,等.基质金属蛋白酶和胶原在创伤关节软骨组织中的表达[J].中国组织工程研究,2011,15(20):3636-3640.
[5]甘业华,孟娟红,马绪臣.我国颞下颌关节骨关节炎基因表达调控及雌激素与关节炎性疼痛关系的研究进展[J].中华口腔医学杂志,2012,47(1):26-27.
[6]王燕,王晓荣,宋涛,等.高频超声在评价药物治疗膝骨关节炎病变中的应用价值[J].中国超声医学杂志,2013,29(9):825-828.
[7]康信忠,吴启富,王康惠,等.中西医结合治疗类风湿关节炎疗效及其对糖皮质激素受体的影响[J].中国中西医结合杂志,2010,30(12):1261-1264.
[8]杨文英,张文丽,罗应伟.颞下颌关节骨关节炎动物模型的研究进展[J].国际口腔医学杂志,2015,42(6):677-680.
[9]阴健,廖爽,张伟华,等.关节腔内注射氨基葡萄糖治疗兔颞下颌关节骨关节炎的组织学研究[J].国际口腔医学杂志,2014,39(1):36-39.
[10]满城,蒋练,徐凡.rh IL-1Ra抑制大鼠颞下颌关节骨关节炎软骨破坏的研究[J].实用口腔医学杂志,2017,33(4):442-446.
[11]兰玲莉,张伟娜,蒋扬眉,等.Notch信号通路在颞下颌骨关节炎中的表达[J].口腔疾病防治,2017,25(3):143-152.
[12]Jabao!nska-Trypu!c A,Matejczyk M,Rosochacki S.Matrix metalloproteinases(MMPs),the main extracellular matrix(ECM)enzymes in collagen degradation,as a target for anticancer drugs[J].J Enzyme Inhib Med Chem,2016,23(8):177-183.
[13]Patterson J,Hubbell JA.Enhanced proteolytic degradation of molecularly engineered PEG hydrogels in response to MMP-1 and MMP-2[J].Biomaterials,2010,31(30):7836-7845.
[14]李干,李奇,林荔军,等.IL-1、MMP-1及TIMP-1在兔骨性关节炎模型滑膜、软骨中的表达[J].实用医学杂志,2011,27(15):2721-2723.
[15]王晓倩,李鑫,郭建生,等.金凤颗粒对痛风性关节炎兔关节软骨MMP-1/TIMP-1表达影响[J].中国免疫学杂志,2015,31(6):774-777.
[16]Eba H,Murasawa Y,Iohara K,et al.The anti-inflammatory effects of matrix metalloproteinase-3 on irreversible pulpitis of mature erupted teeth[J].Plos One,2012,7(12):52523-52530.
[17]Valimaki J,Uusitalo H.Matrix metalloproteinases(MMP-1,MMP-2,MMP-3 and MMP-9,and TIMP-1,TIMP-2 and TIMP-3)and markers for vascularization in functioning and non-functioning bleb capsules of glaucoma drainage implants[J].Acta Ophthalmol,2015,93(5):450-456.
[18]贺占坤,沈杰威.MMP-2、MMP-3、MMP-9和TIMP-1评价膝关节骨性关节炎的临床研究[J].重庆医学,2013,42(32):3872-3874.
[19]李国华,刘德群,刘会仁.大鼠骨骼肌缺血后MMP-9的变化与骨骼肌损伤和重塑的关系[J].安徽医科大学学报,2012,47(11):1308-1311.
[20]石伟哲,肖海军,薛锋,等.基质金属蛋白酶9在大鼠跟腱异位骨化模型中的动态变化研究[J].中国修复重建外科杂志,2014,28(9):116-120.