L-单甲基-精氨酸对兔实验性膝关节骨关节炎Caspases-3、Bax和Bcl-2的影响  

Effect and significance of NG-monomethyl-L-arginine on Caspases-3,Bax and Bcl-2 of rabbits with knee osteoarthritis

作　　者：芦北极[1] 王贵清[1] 查振刚[1] 杨立群[1] 黎绍华[1] 谭小波[1] 

机构地区：[1]暨南大学附属清远医院骨科,广东清远511500

出　　处：《中国医院药学杂志》2013年第12期946-950,共5页Chinese Journal of Hospital Pharmacy

摘　　要：目的:阐明L-单甲基-精氨酸(L-NG-monomethyl-arginine,L-NMMA)对兔实验性膝关节骨关节炎关节软骨细胞凋亡的影响机制。方法:选取30只实验性大白兔随机分为正常组(A组)、模型组(B组)、L-NMMA低剂量治疗组(C组)、L-NMMA中剂量治疗组(D组)和L-NMMA高剂量治疗组(E组),总共5个组,每组6只。采用原位凋亡染色(TUNEL)和流式细胞术检测软骨细胞的凋亡状况;采用蛋白质印迹(western-bloting)法检测各组软骨组织半胱天冬氨酸蛋白酶3(Caspases-3)的表达;采用免疫组织化学方法检测软骨组织Bcl-2相关X蛋白(Human Bcl-2 associated X protein,Bax)和重组人B细胞淋巴瘤因子2(B-Cell Lymphoma-2,Bcl-2)的表达。结果:(1)模型组兔关节软骨细胞的凋亡率和凋亡指数明显高于正常组(P<0.01),且随着L-NMMA治疗浓度的提高,凋亡率和凋亡指数明显下降(P<0.05);(2)模型组兔关节软骨组织Caspases-3的表达水平明显高于正常组(P<0.01),随着L-NMMA治疗浓度提高,Caspases-3表达水平明显下降(P<0.05);(3)模型组兔关节软骨组织Bax的表达水平明显高于正常组(P<0.01),随着L-NMMA治疗浓度提高,Bax的表达水平明显下降(P<0.05);模型组兔关节软骨组织Bcl-2的表达水平与正常组无明显差异性(P>0.05),随着L-NMMA治疗浓度的提高,Bcl-2的表达水平明显上升(P<0.05);随着L-NMMA治疗浓度的提高,Bcl-2/Bax数值明显上升(P<0.05)。结论:L-NMMA可明显降低关节软骨组织Caspases-3的表达水平,可能通过上调Bcl-2/Bax的数值而抑制关节软骨细胞的凋亡,从而为发现阻止OA病变进展的新治疗方法提供实验依据。OBJECTIVE To explore the effects of L-NMMA on the apoptosis of articular cartilage cell in experimental osteoarthritis from rabbit knees and its mechanism of action.METHODS 30 experimental rabbits were randomly divided into five groups： normal group（A）,model group（B）,L-NMMA low-dose treatment group（C group）,L-NMMA middle-dose group（D group） and L-NMMA high-dose treatment group（E group）.In situ apoptosis staining（TUNEL） and flow cytometry was applied to detect the chondrocyte apoptosis;Western-blotting method was used to detect the expression of cartilage Caspases-3;immunohistochemistry was used to detect the expression of cartilage Bax and Bcl-2.RESULTS 1.The apoptosis rate and apoptotic index of rabbit articular chondrocytes in the model group were significantly higher than those in the normal group（P0.01）,and with the increasing treatment concentration of NG-monomethyl-L-arginine arginine,the rate of apoptotic and apoptosis index decreased significantly（P0.05）.2.The expression level of rabbit articular cartilage Caspases-3 in the model group was significantly higher than that in the normal group（P0.01）,and with the increasing treatment concentration of NG-monomethyl-L-arginine,the expression level of Caspases-3 significantly decreased（P0.05）.3.The expression level of rabbit articular cartilage Bax in model group was significantly higher than that in the normal group（P0.01）,and with the increasing treatment concentration of NG-monomethyl-L-arginine,the expression level of Bax significantly decreased（P0.05）;The expression level of rabbit articular cartilage Bcl-2 in the model group showed no significant difference compared with the normal group（P0.05）,and with the increasing treatment concentration of NG-monomethyl-L-arginine,the Bcl-2 expression levels significantly increased（P0.05）;and with the increasing treatment concentration of NG-monomethyl-L-arginine,the numerical value of Bcl-2/Bax significantly increased（P0.05）.CONCLUSION NG-monomethyl-L-argi

关 键 词：L-单甲基-精氨酸 膝关节骨关节炎 关节软骨 凋亡 

分 类 号：R966[医药卫生—药理学]