Ac-SDKP对矽肺大鼠磷酸化热休克蛋白27/SNAI1途径的调节作用  

Regulatory effect of Ac-SDKP on phosphorylated heat shock protein 27/SNAI1 pathway in silicotic rats

作　　者：曹伟 姚珊珊 巩海波 朱丽艳 苗智颖 邓海静 Cao Wei;Yao Shanshan;Gong Haibo;Zhu Liyan;Miao Zhiying;Deng Haijing(School of Basic Medical Sciences,North China University of Science and Technology,Hebei Key Laboratory for Chronic Diseases,Tangshan 063210,China)

机构地区：[1]华北理工大学基础医学院,河北省慢性疾病基础医学重点实验室,唐山063210

出　　处：《中华劳动卫生职业病杂志》2022年第2期90-96,共7页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金(81302395、31800715);河北省高等学校科学技术研究重点项目(ZD2020340);华北理工大学博士科研启动项目;河北省自然科学基金培育基金;华北理工大学杰出青年基金项目(JQ201708)。

摘　　要：目的研究抗纤维化四肽N-乙酰基-丝氨酰-天冬氨酰-赖氨酰-脯氨酸(Ac-SDKP)对矽肺大鼠纤维化肺组织中磷酸化热休克蛋白27(P-HSP27)及锌指家族核转录因子1(SNAI1)表达的调控,以探讨Ac-SDKP抗矽肺纤维化作用。方法于2014年12月,采用一次性支气管灌注二氧化硅(SiO_(2))粉尘的方法制备大鼠矽肺动物模型,选取80只SPF级健康成年Wistar大鼠,根据随机数字表法将大鼠分为8组,每组10只。模型对照4周组(饲养4周)、模型对照8周组(饲养8周):支气管灌注生理盐水1.0 ml/只;矽肺模型4周组(饲养4周)、矽肺模型8周组(饲养8周):支气管灌注50 mg/ml的SiO_(2)混悬液1.0 ml/只;单独Ac-SDKP给药4周组(饲养4周)、单独Ac-SDKP给药8周组(饲养8周):Ac-SDKP 800μg·kg^(-1)·d^(-1)腹腔泵给药;Ac-SDKP预防治疗组:Ac-SDKP 800μg·kg^(-1)·d^(-1)给药48 h后,支气管灌注SiO_(2)混悬液1.0 ml/只,饲养8周;Ac-SDKP抗纤维化治疗组:支气管灌注SiO_(2)混悬液1.0 ml/只4周后,给予Ac-SDKP 800μg·kg^(-1)·d^(-1)治疗4周。蛋白免疫印迹(Western blotting)检测各组P-HSP27、SNAI1、α-平滑肌肌动蛋白(α-SMA),以及Ⅰ型和Ⅲ型胶原蛋白的表达,免疫组织化学技术检测P-HSP27和SNAI1的表达,激光共聚焦显微镜检测P-HSP27和α-SMA共定位表达。结果与模型对照组比较,矽肺模型组大鼠矽肺纤维化区域P-HSP27、SNAI1、α-SMA、Ⅰ型和Ⅲ型胶原蛋白表达增强,差异均有统计学意义(P<0.05)。给予Ac-SDKP干预后,与矽肺模型8周组比较,Ac-SDKP预防和抗纤维化治疗组大鼠P-HSP27、SNAI1、α-SMAⅠ型和Ⅲ型胶原蛋白表达明显降低,差异均有统计学意义(P<0.05)。而单独Ac-SDKP给药组与模型对照组P-HSP27、SNAI1、α-SMA、Ⅰ型和Ⅲ型胶原蛋白表达无明显变化,差异均无统计学意义(P>0.05)。激光共聚焦结果显示,矽肺模型组大鼠肺组织表达P-HSP27和α-SMA的阳性细胞多于模型对照组;与矽肺模型组比较,Ac-SDKP预防和抗纤维化Objective To study the effect of anti-fibrotic tetrapeptide N-acetyl-seryl-aspartyl-lysyl-proline(Ac-SDKP)on phosphorylated heat shock protein 27(P-HSP27)and zinc finger family transcriptional repressor 1(SNAI1)expression to explore the anti-silicosis fibrosis effect of Ac-SDKP.Methods In December 2014,the rat silicosis animal model was prepared by one-time bronchial infusion of silicon dioxide(SiO_(2))dust.80 SPF healthy adult Wistar rats were selected,and the rats were divided into 8 groups according to the random number table method,10 in each group.Model control group for 4 weeks(feeding for 4 weeks),model control group for 8 weeks(feeding for 8 weeks):bronchial perfusion with normal saline 1.0 ml per animal.Silicosis model group for 4 weeks(feeding for 4 weeks)and silicosis model group for 8 weeks(feeding for 8 weeks):bronchial perfusion of 50 mg/ml SiO_(2)suspension 1.0 ml per animal.Ac-SDKP administration group for 4 weeks(feeding for 4 weeks),Ac-SDKP administration group for 8 weeks(feeding for 8 weeks):Ac-SDKP 800μg·kg^(-1)·d^(-1)was administered by intraperitoneal pump.Ac-SDKP preventive treatment group:48 h after Ac-SDKP 800μg·kg^(-1)·d^(-1)administration,bronchial perfusion of SiO_(2)suspension 1.0 ml per animal,raised for 8 weeks.Ac-SDKP anti-fibrosis treatment group:after bronchial perfusion of 1.0 ml of SiO_(2)suspension for 4 weeks,Ac-SDKP 800μg·kg^(-1)·d^(-1)was administered for 4 weeks.Western blotting was used to detect the expression of P-HSP27,SNAI1,α-smooth muscle actin(α-SMA),and collage typeⅠandⅢin each group.The expression of P-HSP27 and SNAI1 was detected by immunohistochemistry,and the co-localized expression of P-HSP27 andα-SMA was detected by laser confocal microscopy.Results Compared with the model control group,the expressions of P-HSP27,SNAI1,α-SMA,and collage typeⅠandⅢin the silicosis fibrosis area of the rats in the silicosis model group were enhanced,and the differences were statistically significant(P<0.05).After Ac-SDKP intervention,compared with silicosis

关 键 词：矽肺 大鼠 N-乙酰基-丝氨酰-天冬氨酰-赖氨酰-脯氨酸(Ac-SDKP) 上皮-间质转化 热休克蛋白质类 锌指家族核转录因子1(SNAI1) 

分 类 号：R135.2[医药卫生—劳动卫生]