睡眠剥夺抑制大鼠血管生物钟因子CRY1表达促进血管衰老的实验研究

Experimental study on sleep deprivation inhibiting clock gene CRY1 expression in vascular tissue and promoting vascular senescence

摘要:

目的]探讨睡眠剥夺与血管衰老之间的关系及可能存在的机制。 [方法]雄性SD大鼠分为对照组、衰老模型组、睡眠剥夺组及睡眠剥夺+衰老组,每组各6只。改良水平台法剥夺大鼠睡眠时长。衰老相关β半乳糖苷酶(SA-β-Gal)染色检测大鼠血管衰老状况。实时荧光定量PCR(RT-qPCR)、Western blot及免疫组织化学法测定各组大鼠血管组织中肿瘤抑制蛋白p53、沉默信息调节因子1(SIRT1)及生物钟因子隐花色素1(CRY1)的表达水平。 [结果]与对照组相比,衰老模型组大鼠血管组织中SA-β-Gal染色加深,p53水平升高,SIRT1水平降低。与对照组相比,睡眠剥夺组、睡眠剥夺+衰老组大鼠血管组织存在与衰老模型组相似的改变,包括SA-β-Gal染色加深,p53水平升高,SIRT1水平降低。此外,相较于对照组,睡眠剥夺组大鼠血管组织中CRY1水平降低,睡眠剥夺+衰老组大鼠血管组织中仅CRY1 mRNA水平降低。与衰老模型组相比,睡眠剥夺+衰老组大鼠血管组织中的SA-β-Gal染色加深,p53水平升高,SIRT1水平降低,CRY1 mRNA水平降低。 [结论]睡眠剥夺可促进血管衰老相关因子表达,其机制可能与抑制血管组织中CRY1的表达有关。

Abstract:

Aim To investigate the relationship between sleep deprivation and vascular aging, as well as the underlying mechanisms. Methods Male Sprague-Dawley rats were divided into control group, senescence group, sleep deprivation group, and sleep deprivation+senescence group, with 6 rats in each group. The modified level table method deprived rats of sleep duration. Senescence-associated β-galactosidase (SA-β-Gal) staining was used to detect the senescence status of rat vascular tissue. The mRNA and protein expression of tumor suppressor protein p53, silent information regulator 1 (SIRT1) and clock gene cryptochrome 1 (CRY1) was detected by real-time fluorescence quantification PCR (RT-qPCR), Western blot and immunohistochemistry. Results Compared with the control group, the intensity of SA-β-Gal staining was increased in the vascular tissues of the senescence group rats, the expression level of p53 was elevated , the expression level of SIRT1 was decreased. Similar changes were observed in the sleep deprivation group and the sleep deprivation+senescence group, including intensified SA-β-Gal staining, elevated p53 levels, and reduced SIRT1 levels in vascular tissues. Additionally, compared with the control group, the sleep deprivation group showed reduced CRY1 levels in vascular tissues, while only CRY1 mRNA levels were reduced in the sleep deprivation+senescence group. Furthermore, compared with the senescence group, the sleep deprivation+senescence group exhibited intensified SA-β-Gal staining, increased p53 level, decreased SIRT1 level, and reduced CRY1 mRNA level in vascular tissues. Conclusion Sleep deprivation may promote the expression of vascular aging-related factors, potentially through the inhibition of CRY1 expression in vascular tissues.