慢性睡眠剥夺对大鼠颞下颌关节ERK信号通路的影响

发布时间：2018-07-22 12:16

【摘要】：目的：探讨细胞外信号调节激酶(ERK)通路在慢性睡眠剥夺(CSD)引起大鼠颞下颌关节(TMJ)结构变化中的作用。方法： 270只雄性Wistar大鼠随机分为3组：慢性睡眠剥夺组(CSD)、慢性睡眠剥夺+U0126(ERK抑制剂)组(U0126)、对照组(CON),每组90只。每组根据试验时间不同分别分为3个亚组：7天(7d)、14天(7d)、21天(21d)组,每个亚组30只。参考改良多平台法(MMPM)建立大鼠的慢性睡眠剥夺模型。通过矿场试验及测定大鼠血清中皮质酮(CORT)和促肾上腺皮质激素(ACTH)水平,评估模型的效果；分别通过HE染色和扫描电镜观察大鼠TMJ的病理和超微结构变化；通过Western Blot测定髁突软骨ERK和磷酸化ERK (P-ERK)的蛋白表达变化；通过Western Blot和实时荧光定量PCR检测MMP-1、MMP-3和MMP-13的表达变化。结果： CSD组的大鼠旷场试验中,CSD各组大鼠的水平得分及垂直得分均明显高于CON各组大鼠(P0.05)；CSD各组大鼠的血清CORT和ACTH水平与CON组相比明显升高(P0.05)。 HE染色和扫描电镜结果显示,CSD组的大鼠TMJ出现病理性的改变,U0126组的大鼠TMJ关节破坏程度减轻。与CON组比较,CSD组P-ERK的蛋白表达水平明显升高(P0.01),总ERK水平无明显变化(P0.05),MMP-1、MMP-3和MMP-13的mRNA转录和蛋白表达水平明显升高(P0.05)。与CSD组比较,U0126组P-ERK的蛋白表达明显降低(P0.01),总ERK水平无明显变化(P0.05),MMP-1、MMP-3和MMP-13的mRNA转录和蛋白表达水平下降(P0.05)。结论：改良多平台法是一种比较理想的使大鼠处于慢性睡眠剥夺的应激状态模型。慢性睡眠剥夺能够引起大鼠颞下颌关节髁突软骨的病理性变化。这种病理性变化可能是引起颞下颌关节功能紊乱综合征的病理性基础。慢性睡眠剥夺能够激活大鼠颞下颌关节髁突软骨的ERK通路,通过上调MMP-1、MMP-3和MMP-13的表达水平,继而引起大鼠颞下颌关节的病理性变化。ERK通路抑制剂能够减轻这种病理性变化。这些说明ERK通路在慢性睡眠剥夺引起的颞下颌关节病理变化中起关键作用。
[Abstract]:Aim: to investigate the role of extracellular signal-regulated kinase (ERK) pathway in the structural changes of temporomandibular joint (TMJ) induced by chronic sleep deprivation (CSD) in rats. Methods: 270 male Wistar rats were randomly divided into three groups: chronic sleep deprivation group (CSD), chronic sleep deprivation U0126 (ERK inhibitor) group (U0126) and control group (Con) with 90 rats in each group. Each group was divided into 3 subgroups: 7 days (7 days), 14 days (7 days) and 21 days (21 days), with 30 rats in each subgroup. A rat model of chronic sleep deprivation was established by reference to the modified multi-platform method (MMPM). The effect of the model was evaluated by field test and determination of corticosterone (Cort) and adrenocorticotropic hormone (ACTH) in serum of rats, and the pathological and ultrastructural changes of TMJ were observed by HE staining and scanning electron microscope, respectively. The protein expression of ERK and phosphorylated ERK (P-ERK) in condylar cartilage was detected by Western Blot, and the expression of MMP-3 and MMP-13 in condylar cartilage was detected by Western Blot and real-time fluorescent quantitative PCR. Results: the horizontal and vertical scores in CSD group were significantly higher than those in Con group (P0.05). The levels of serum Cort and ACTH in CSD group were significantly higher than those in Con group (P0.05). The pathological changes of TMJ in CSD group were observed by HE staining and scanning electron microscope. The degree of joint destruction of TMJ in U0126 group was reduced. Compared with the Con group, the expression of P-ERK protein in CSD group was significantly higher (P0.01), but the total ERK level had no significant change (P0.05) the mRNA transcription and protein expression level of MMP-1MMP-3 and MMP-13 were significantly increased (P0.05). Compared with CSD group, the expression of P-ERK protein in U0126 group was significantly lower (P0.01), but the total ERK level had no significant change (P0.05) the mRNA transcription and protein expression of MMP-1MMP-3 and MMP-13 decreased (P0.05). Conclusion: the modified multi-platform method is an ideal stress state model for chronic sleep deprivation in rats. Chronic sleep deprivation can cause pathological changes of condylar cartilage of temporomandibular joint in rats. This pathological change may be the pathological basis of temporomandibular joint dysfunction syndrome. Chronic sleep deprivation can activate ERK pathway of condylar cartilage of temporomandibular joint in rats. By up-regulating the expression of MMP-1, MMP-3 and MMP-13, the pathological changes of temporomandibular joint can be induced by chronic sleep deprivation. ERK pathway inhibitor can attenuate this pathological change. These results suggest that ERK pathway plays a key role in the pathological changes of temporomandibular joint induced by chronic sleep deprivation.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R740;R782

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