Effect Of GGCX On Rabbit Cartilage Cell Apotosis In Osteoarthritis And Expression Of IL-1β、TNF

Objective:To investigate the effect of GGCX mediated by lentivirus on rabbit cartilage cell apotosis in osteoarthritis and the expression of IL-1β, TNF.Methods:1、12 male Japanese big ear rabbits were choosed for knee osteoarthritis model induced by anterior cruciate ligament transaction. After 6 weeks, the constructed model rabbits were sacrificed to isolate and culture articular chondrocytes. Type II collagen fiber staining and Alcian Blue staining method were used to indentify the cultured chondrocytes.2 、The cultured chondrocytes were divided randomly into three groups: the blank group(no treatment), the negative group(transduct by lentivirus without GGCX), and the positive group(transduct by lentivirus with GGCX). The expression of GGCX, type II collagen, TNF, IL-1β at mRNA and protein levels in different groups was detected by qRT-PCR and Western blot。3、The apoptosis ratios of cells in different groups were analysised by Annexin V-FITC & PI method.Results:1、Osteoarthritis cartilage cells at passage 1 were predominantly consisted of spolygonal and flat cells with a plurality of protrusions. After type II collagen fiber staining, cells and extracellular matrix were stained yellow or brown. After Alcian Blue staining, cells showed blue.2、 The results of qRT-PCR and Western blot showed that the expression of GGCX and type II collagen at protein and mRNA levels was significantly increased in the positive group compared with the blank group and the negative control group,the difference was statistically significant(P < 0.05); however, the blank group and the negative control group have no significant difference(P > 0.05). The expression of TNF 、 IL-1β at protein and mRNA levels was significantly decreased in thepositive group compared with the blank group and the negative group, the difference was statistically significant(P < 0.05); moreover, the blank control group and the negative control group have no significant difference(P > 0.05).3、The apoptosis ratios of cells analysised by Annexin V-FITC & PI method showed that the apoptosis ratios of cells in the positive group was(17.833±2.577)%,revealing a significantly decrease compared with the blank group with(25.467±2.511)% and the negative group with(26.167±2.616)%, the difference was statistically significant(P < 0.05). The blank group and the negative group have no significant difference(P > 0.05).Conclusions:1 、 The knee osteoarthritis model was constructed sucessefully induced by anterior cruciate ligament transaction, and articular chondrocytes can be isolated from the knee osteoarthritis model.2、Transducting GGCX by lentivirus can significantly increase the expression of GGCX, type II collagen in chondrocytes.3、Transducting GGCX by lentivirus can significantly decrease the expression of TNF, IL-1β in chondrocytes.4、Transducting GGCX by lentivirus can inhibit the apoptosis of cartilage cells.