Experimental Study On D-AIa2GIP Against Abnormal Extracellular Matrix Metabolism In Osteoarthritis Rat

Objective:Observation glucose-dependent incretin analogs DAIa2 GIP antagonistic osteoarthritis model rat extracellular matrix metabolism.Methods:The 18 male SD rats(purchased from Experimental Animal Center of Shanxi Medical University)were divided into three groups(sham-operated group,model group,the treatment group),n = 6:the model group,DAIa2 GIP treatment group after cutting the front left and right posterior cruciate ligament,medial meniscus resection;sham operation group incision around the capsule of the knee,without cutting the anterior cruciate ligament and the medial meniscus;modeling complete after 8 weeks,the model group,DAIa2 GIP treatment group were injected with 25 nmol / kg glucose-dependent incretin analogs DAIa2 GIP,2 times / week,8 injections weeks;sham group was given intraperitoneally injected with equal volume of saline.After the 16 th week,sudden death(1)SD rats in each group,each group of the knee joint and remove substantially ICRS score,and the knee joint of the rats were Safranin "O"-Fast Green staining,observation of rats the structural characteristics of the knee joint cartilage;(2)mitochondrial membrane potential assay kit(JC-1)detected by the transition of mitochondrial membrane potential fluorescent colors change.(3)synovial fluid extraction SD rats of each group,ELISA Detection of rats taken knee MMP-13 and IL-1? factor;(4)Western blot measuring cytochrome c,a change in type II collagen.Results:(1)(1)generally Rating: compared with the sham group(15.50 � 0.83),slightly lower scoring model treated group(13.67 � 0.81,P> 0.05),significantly lower scores in model group(8.00 � 0.89,P <0.05),the model treatment group compared with the model group(P <0.05).(2)Safranin O-fast green staining: sham proteoglycan expression of multiple uniform,relatively normal thickness cartilage;compared with sham group,model group,the expression of proteoglycan less uniform groups thereof and cartilage thinner thickness,degree of fibrosis / range are large and heavy;treatment group and the model used proteoglycan expression after treatment DAla2 GIP substantially,even still,cartilage thickness increase over the hand model group.(2)Mitochondrial membrane potential change: comparing the treatment group model membrane potential change ranges slightly sham group(12.80% � 2.91%)(15.68% � 1.19%,P> 0.05),membrane potential change range of the model group increased significantly(21.06% �0.87%,P <0.05),model group and the treatment group compared model(P <0.05).(3)ELISA Detection:(1)IL-1?: sham group(369.36 � 33.42 pg / ml)were compared,IL-1? treatment group content model is slightly higher(387.24 � 24.05 pg / ml,P>0.05),IL-1? model group It was significantly increased(896.57 � 76.19 pg / ml,P<0.05);model treatment group compared with the model group(P <0.05).(2)MMP-13:compared with the sham group(500.83 � 65.38 pg / ml),the content of the model of MMP-13 treatment group increased slightly(562.13 � 79.23 pg / ml,P> 0.05),the content of the model group significantly MMP-13 increase(803.25 � 84.47 pg / ml,P<0.05),treating the model group compared with the model group(P <0.05).(4)Westernblot detection of tissue samples:(1)Cyt-c: compared with the sham group(100%� 3%),a model for treatment group Cyt-c content was slightly higher(140% � 33%,P> 0.05),model group Cyt-c It was significantly increased(188% � 16%,P <0.05);model treatment group compared with the model group(P <0.05).(2)col-II: sham operation group(98% � 2%),col-II content model treatment group decreased slightly(92% � 16%,P> 0.05),model group col-II significantly decreased(39% � 7%,P<0.05);model treatment group compared with the model group(P <0.05).Conclusion:(1)DAla2GIP can inhibit cartilage damage abnormal cell mitochondrial membrane potential depolarization release trigger Cyt C;(2)DAla2GIP reversible disturbance of extracellular matrix osteoarthritis cells,such as IL-1?,col-II,MMP-13,to further protect cartilage.