| Objective: To investigate whether there is a difference in the expression of lnc RNAHOTAIR in articular cartilage of patients with knee osteoarthritis compared with normal controls,and to explore the role of HOTAIR expression in IL-1β-mediated chondrocyte apoptosis and extracellular matrix degradation.Methods:The medial knee condylar cartilage of patients with knee osteoarthritis(OA)undergoing total knee arthroplasty(TKA)in our hospital was taken as the experimental group(OA group),and the articular cartilage of patients without arthritis was taken as the control group(Nor group).The expression difference of lnc RNAHotair in articular cartilage of the two groups was analyzed by RT-PCR.The rat model of osteoarthritis was established and divided into 4W,6W,and 8W according to the sampling time.The difference of lnc RNAHOTAIR in articular cartilage of osteoarthritis rats(OA group)and normal rats(Nor group)was analyzed,and the difference of HOTAIR expression in articular cartilage of osteoarthritis rats(4W,6W,and 8W)was analyzed by intra-group comparison.Rat chondrocytes were cultured and stimulated with IL-1β to study the changes of HOTAIR in chondrocytes and to further clarify whether HOTAIR could achieve overexpression through IL-1βstimulation.The experimental designs were the Con group,IL-1β group,siHOTAIR group,and IL-1β+ si-HOTAIR group.Respectively,representing blank control,IL-1β was added for intervention,si RNA of the HOTAIR gene was added for interference,and IL-1β and si-HOTAIR were added together for intervention.CCK-8 assay and flow cytometry were used to determine whether HOTAIR gene expression could affect the activity and apoptosis of IL-1βmediated chondrocytes.Experiments by Western blot method different experimental expression of type collagen Ⅱ.To determine whether the HOTAIR gene is affected by IL-1β and interference of chondrocyte cartilage matrix synthesis.Results:1.The human cartilage experiment showed that the expression of HOTAIR was significantly up-regulated in the OA group compared with the Nor group.Animal experiments showed that the expression of HOTAIR in the oa group was also up-regulated compared with that in the nor group,and with the change of modeling time,the expression of HOTAIR showed an upward trend.2.To determine whether HOTAIR was affected by IL-1β,we designed the con group and IL-1β group,and the results showed that HOTAIR expression was up-regulated in the IL-1β group.3.The results of the CCK-8 experiment showed that compared with the Con group,the activity of rat articular chondrocytes was decreased in the IL-1βgroup,and there was no significant change in the activity of rat articular chondrocytes in the si-HOTAIR group.Compared with the IL-1β group,the activity of articular chondrocytes increased in the IL-1β+ si-HOTAIR group.4.Flow cytometry showed that compared with the Con group the apoptosis level of the IL-1β group was significantly increased,but there was no significant difference in the si-HOTAIR group.Compared with the IL-1βgroup,the apoptosis level of the IL-1β+ si-HOTAIR group was significantly decreased.5.Through the Western-blot method,the experimental results show that compared with Con group,IL-1 beta group cut collagen type Ⅱ expression,siHotair collagen type Ⅱ protein expression did not change;Compared with IL-1beta group,IL-1 beta + si-Hotair group showed an increased collagen typeⅡprotein expression.Conclusion: HOTAIR expression is up-regulated in articular cartilage of knee arthritis,and this regulation is mediated by IL-1β.Hotair expression is mediated by IL-1beta,making reduced articular cartilage cell vitality,increased apoptosis,collagen tpyeⅡ expression is reduced,and however,after the interference of Hotair,cartilage cell-mediated by IL-1 beta vitality increased,apoptosis reduced,collagen typeⅡexpression increased;These results indicate that HOTAIR is an important factor in the development of arthritis and can exert its negative effects as a downstream pathway in IL-1β-mediated arthritis. |
PDF Full Text Request