凉血消风汤对HaCaT细胞中组蛋白H4乙酰化及其相关酶的干预机制研究  

Study on Intervention Mechanism of Liangxue Xiaofeng Decoction on Acetyl-Histone H4 and Related Enzymes in HaCaT Cells

作　　者：王文欢 王亚翠 贾瑞璇 王红梅[4] WANG Wenhuan;WANG Yacui;JIA Ruixuan;WANG Hongmei(Dalian Dermatosis Hospital,Dalian Liaoning 116021,China;Sichuan Science City Hospital,Mianyang Sichuan 621999,China;Tianjin Emergency Center,Tianjin 300120,China;Tianjin Academy of Traditional Chinese Medicine Affiliated Hospital,Tianjin 300120,China)

机构地区：[1]大连市皮肤病医院,辽宁大连116021 [2]四川省科学城医院,四川绵阳621999 [3]天津市急救中心,天津300120 [4]天津市中医药研究院附属医院,天津300120

出　　处：《新中医》2025年第5期202-208,共7页New Chinese Medicine

基　　金：大连市医学科学研究计划项目(21Z11024)。

摘　　要：目的:研究凉血消风汤对人永生化角质形成细胞(HaCaT)中组蛋白H4乙酰化(Acetyl-Histone H4)及组蛋白乙酰基转移酶1(HAT1)、乙酰化转移酶P300、去乙酰化酶sirtuin(SIRT)3、SIRT5的干预作用,进而探讨乙酰化修饰参与银屑病发病的作用机制。方法:采用MTT法检测凉血消风汤对HaCaT细胞存活率的影响,设立HaCaT细胞空白对照组,凉血消风汤组(中药高、中、低浓度组)、雷公藤组、姜黄素组。用蛋白质印迹检测HaCaT细胞加药前后Acetyl-Histone H4及其相关酶HAT1、P300、SIRT3、SIRT5的蛋白表达,实时荧光定量PCR测定相关酶HAT1、P300、SIRT3、SIRT5 mRNA的表达。结果:根据MTT检测结果,凉血消风汤分别选2、1、0.25 ng/mL作为中药高、中、低浓度组的给药剂量,雷公藤组为2.5μg/mL,姜黄素组为5μmol/L。与空白对照组比较,雷公藤组及中药高、中浓度组中Acetyl-Histone H4、HAT1、P300、SIRT3的蛋白表达均增加(P<0.05),SIRT5蛋白表达降低(P<0.05)。与空白对照组比较,雷公藤组、中药高浓度组中HAT1、P300、SIRT3的mRNA表达增加,SIRT5 mRNA表达降低(P<0.05);中药中浓度组中HAT1、P300 mRNA表达增加(P<0.05),SIRT5 mRNA表达降低(P<0.05);中药低浓度组中SIRT5 mRNA表达降低(P<0.05);姜黄素组HAT1 mRNA表达降低(P<0.05)。结论:凉血消风汤可以上调HaCaT细胞中Acetyl-Histone H4、乙酰化酶HAT1、P300及去乙酰化酶SIRT3的表达水平,下调SIRT5的表达水平,推测凉血消风汤可能是通过调控组蛋白乙酰化及相关酶来治疗银屑病,乙酰化内稳态改变可能是银屑病发病的一个重要因素。Objective:To investigate the effect of Liangxue Xiaofeng Decoction on Acetyl-Histone H4,histone acetyltransferase 1(HAT1),acetyltransferase P300,deacetylase sirtuin(SIRT)3 and SIRT5 in HaCaT cells,and to further discuss the mechanism of acetylation in the pathogenesis change of psoriasis.Methods:The effect of Liangxue Xiaofeng Decoction on the survival rate of HaCaT cells was detected by MTT method,and the following groups of HaCaT were set up:the blank control group,the Liangxue Xiaofeng Decoction group(high-,medium-and lowconcentration Chinese medicine groups),the Tripterygium group and the Curcumin group.The Acetyl-Histone H4 and the protein expressions of HAT1,P300,SIRT3 and SIRT5 were detected by Western Blot before and after administration,and the mRNA expressions of HAT1,P300,SIRT3 and SIRT5 were detected by real-time fluorescent quantitative PCR.Results:According to the results of MTT test,2,1 and 0.25 ng/mL were selected as the dosages of high-,medium-and low-concentration Chinese medicine groups,respectively,2.5μg/mL in the Tripterygium group and 5μmol/L in the Curcumin group.When compared with those in the blank control group,the protein expressions of Acetyl-Histone H4,HAT1,P300 and SIRT3 in the Tripterygium group,high-and medium-concentration Chinese medicine groups were increased(P<0.05),and the protein expressions of SIRT5 were decreased(P<0.05).When compared with those in the blank control group,mRNA expressions of HAT1,P300 and SIRT3 were increased in the Tripterygium group and the high-concentration Chinese medicine group,and the mRNA expressions of SIRT5 were decreased(P<0.05).The mRNA expressions of HAT1 and P300 in the medium-concentration Chinese medicine group were increased(P<0.05),and the mRNA expression of SIRT5 was decreased(P<0.05).The mRNA expression of SIRT5 in the low-concentration Chinese medicine group was decreased(P<0.05).The mRNA expression of HAT1 in the Curcumin group was decreased(P<0.05).Conclusion:Liangxue Xiaofeng Decoction can up-regulate the expression levels of Acety

关 键 词：银屑病 凉血消风汤 人永生化角质形成细胞 组蛋白乙酰基转移酶1 乙酰化酶P300 去乙酰化酶sirtuin 3 去乙酰化酶sirtuin 5 

分 类 号：R285.5[医药卫生—中药学]