Evaluation of rat osteoarthritis chondrocyte models induced by interleukin-1beta / 中国组织工程研究

ABSTRACT

BACKGROUND:Establishing a chondrocyte model of osteoarthritis is of great significance for further explaining the pathological process of osteoarthritis and evaluating and screening the therapeutic drugs of osteoarthritis. OBJECTIVE:To evaluate the effect of interleukin-1β to induce osteoarthritis models in rat chondrocyte models,thereby providing a reference for further exploration of drug treatment of osteoarthritis. METHODS:Chondrocytes were isolated from the hip cartilage of 3-week-old Sprague-Dawley rats by mechanical shearing and enzymatic digestion,and then identified.Chondrocytes were randomly divided into three groups:control group,5 ng/mL interleukin-1β-induced group,10 ng/mL interleukin-1β-induced group,with induction times of 24 and 48 hours.Chondrocyte proliferation activity was detected by MTT.Real-Time PCR was used to detect the expression of type Ⅱ collagen,Aggrecan,sex-determining region Y-box protein 9(Sox9),matrix metalloproteinase 13,and a disintegrin and metaloproteinase with thrombospondin-like motifs-5(Adamts5)mRNA.Western blot was used to detect the expression of type Ⅱ collagen,Sox9,matrix metalloproteinase 13 and Adamts5. RESULTS AND CONCLUSION:Primary rat chondrocytes were successfully isolated and cultured.Induction of chondrocytes by interleukin-1β at 10 ng/mL for 24 hours could significantly reduce cell proliferation and viability(P＜0.05),while the 5 ng/mL interleukin-1β-induced group required 48 hours of induction to cause a significant decrease in cell proliferation and viability.Real-Time PCR results showed that compared with the control group,5 or 10 ng/mL interleukin-1β induction for 24 and 48 hours significantly reduced the expression levels of type Ⅱ collagen,Aggrecan,Sox9 mRNAs(P＜0.05)and increased the expression levels of matrix metalloproteinase 13 and Adamts5 mRNAs(P＜0.05).Compared with the 10 ng/mL interleukin-1β-induced group,5 ng/mL interleukin-1β induction significantly increased the mRNA expression of matrix metalloproteinase 13 and Adamts5 in chondrocytes after 48 hours of induction(P＜0.05).Western blot results showed that compared with the control group,10 ng/mL interleukin-1β induction for 24 hours and 5 ng/mL interleukin-1β induction for 48 hours significantly decreased the protein expression of type Ⅱ collagen and Sox9 in chondrocytes(P＜0.05),and significantly increased the protein expression of matrix metalloproteinase 13 and Adamts5(P＜0.05).To conclude,compared with 5 ng/mL interleukin-1β,10 ng/mL interleukin-1β may have more obvious effects on chondrocytes for 24 hours,while 5 and 10 ng/mL interleukin-1β have similar effects after 48 hours of intervention.