慢病毒介导RNAi沉默IL-1 Ⅰ型受体在大鼠骨关节炎MAPK信号转导中的作用

摘要

Objective To observe the effects of lentivirus-mediated RNA interference silencing IL-1 type Ⅰ receptor on the expression of mitogen-activated protein kinase (MAPK) in a rat model of osteoarthritis (OA), and to explore the role of IL-1 and MAPK in OA. Methods Forty SD rats were randomly divided into four groups(A-D, each group: n = 10). The rats (A-C group) were underwent unilateral anterior cruciate ligament transaction and partial excision of the medial meniscus. At day 7 and 9 after operation, the rats of group A were respectively received 40 μL of lentivirus-mediated RNAi silencing IL-1 type Ⅰ receptor by intra-articular injection, while the rats of group B were received lentivirus-mediated RNAi non-silencing IL-1 type Ⅰ receptor, and the rats of group C were received saline. The rats of group D were taken for normal control. All rats were sacrificed four weeks after the surgery. The knees were harvested to observe macropathologic changes of the joint cartilage. ERK1/2, JNK1/2 and p38 proteins in the cartilage were detected by Western blot. Results Cartilage degradation in group A was milder than that in group B and C (P<0.05),but was worse compared to group D (P<0.05). Level of p38 expression in cartilage in group A was lower than that in group B and C (P<0. 05), but had no significant difference compared to group D (P>0.05). Levels of ERK1/2 and JNK1/2 expressions in cartilage in group A were lower than that in group B and C(P<0.05),but was significantly increased compared to group D(P<0.05). Conclusions Lentivirus-mediated RNAi silencing IL-1 type Ⅰ receptor can inhibit the expression of MAPK, in particular,p38 protein was strongly inhibited.%目的 观察慢病毒介导的RNAi沉默IL-1 Ⅰ型受体对实验性大鼠膝骨关节炎(osteoarthritis,OA)中的丝裂素原激活的蛋白激酶(mitogen-activated protein kinase,MAPK)途径的影响,探讨IL-1和MAPK在OA中的作用.方法 40只SD大鼠随机分为A～D 4组,每组10只.A～C组行单侧前交叉韧带和内侧半月板前1/3切除术,分别于术后第7和9天,A组关节腔中注射慢病毒介导IL-1 Ⅰ型受体有效RNA干扰片段,B组注射慢病毒介导IL-1 Ⅰ型受体无效RNA干扰片段,C组单纯注射生理盐水.D组为正常动物不做处理.术后4周处死动物,对关节软骨的状况进行评分.取膝关节软骨组织.用Western blot方法检测软骨组织中p38、ERK1/2和JNK1/2的蛋白质表达水平.结果 大体观察显示:A组软骨退行性变显著轻于B、C组(P<0.05),但高于D组(P<0.05).Western blot显示,A组p38蛋白质表达水平显著低于B、C组(P<0.05),与D组没有显著性差异(P>0.05).A组ERK1/2和JNK1/2的表达显著低于B、C组(P<0.05),但要显著高于D组(P<0.05).结论 慢病毒介导RNA干扰沉默IL-1 Ⅰ型受体能抑制MAPK的表达,特别是对其中的p38蛋白质抑制更为明显.