益肾达络饮对DPCPX干预下EAE小鼠H3K9ac、H4K5ac表达的影响  

Effect of Yishen Daluo Decoction on the Expression of H3K9ac and H4K5ac in EAE mice under DPCPX intervention

作　　者：王响 谢名宗 何静文 丁文婧 尚晓玲[1] WANG Xiang;XIE Mingzong;HE Jingwen;DING Wenjing;SHANG Xiaoling(Changchun University of Chinese Medicine,Changchun 130117,China)

机构地区：[1]长春中医药大学,长春130117

出　　处：《中国中医基础医学杂志》2023年第10期1634-1639,共6页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：国家自然科学基金项目(81873216)。

摘　　要：目的观察益肾达络饮在腺苷A1受体(adenosine A1 receptor,A1AR)抑制剂1,3-二丙-8-环戊黄嘌呤(8-cyclopentyl-1,3-dipropylxanthine,DPCPX)干预下对实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)小鼠的治疗作用及对组蛋白H3第9位赖氨酸乙酰化(histone H3 lysine residue 9 acetylation,H3K9ac)、组蛋白H4第5位赖氨酸乙酰化(histone H4 lysine residue 5 acetylation,H4K5ac)的影响。方法将60只C57BL/6J雌性小鼠随机平均分成6组。除正常对照组外,其余组均制作EAE模型。从造模第8天开始,模型组、DPCPX组予0.9%氯化钠溶液灌胃,西药组予醋酸泼尼松混悬液灌胃,中药组、DPCPX+中药组予益肾达络饮药液灌胃,连续给药14 d。每日记录各组小鼠的神经功能评分,运用Western Blot检测各组小鼠脑及脊髓中A1AR、H3K9ac、H4K5ac的表达水平,免疫组化观察脑及脊髓中H3K9ac、H4K5ac的表达情况。结果与正常对照组比较,模型组最高平均神经功能评分与第21天评分均显著升高(P<0.01);与模型组比较,中药组、DPCPX+中药组、西药组最高平均神经功能评分与第21天评分均显著下降(P<0.05)。Western Blot结果显示,与正常对照组比较,模型组脑与脊髓组织中A1AR显著下降(P<0.01),H3K9ac、H4K5ac显著升高(P<0.05);与模型组比较,DPCPX组脑中A1AR显著下降(P<0.05),脑与脊髓中H3K9ac、H4K5ac显著升高(P<0.05);中药组脊髓中A1AR表达量显著升高(P<0.05),脑中H3K9ac、H4K5ac显著下降(P<0.05);DPCPX+中药组脑与脊髓中A1AR显著升高(P<0.05),脑中H3K9ac显著升高(P<0.05),H4K5ac显著下降(P<0.05),脊髓中H4K5ac显著升高(P<0.05);西药组脊髓中A1AR显著升高(P<0.01),脑中H3K9ac显著下降(P<0.01),H4K5ac显著升高(P<0.05);免疫组化显示,与正常对照组比较,模型组脑与脊髓组织H3K9ac、H4K5ac表达量显著升高(P<0.05);与模型组比较,DPCPX组脑与脊髓组织H3K9ac、H4K5ac表达量显著升高(P<0.05),中药组、西药组脑与脊髓组织H3KObjective To observe the effect of Yishen Daluo decoction on the expression of histone H3 lysine residue 9 acetylation(H3K9ac)and histone H4 lysine residue 5 acetylation(H4K5ac)in experimental autoimmune encephalomyelitis(EAE)mice under the intervention of 8-cyclopentyl-1,3-dipropylxanthine(DPCPX)that is the inhibitor of adenosine A1 receptor(A1AR).Methods Sixty C57BL/6J female mice were randomly divided into 6 groups.Except for the normal control(NC)group,the EAE model was made in the other groups.From the 8th day of modeling,the model and DPCPX groups were given 0.9%sodium chloride solution by gavage,the hormone group was given prednisone acetate,the traditional Chinese medicine(TCM)and DPCPX+TCM groups were given Yishen Daluo decoction.The drugs were given continuously for 14 days.The clinical scores of mice were recorded daily.Western Blot(WB)was used to detect A1AR,H3K9ac,and H4K5ac in the brain and spinal cord.The expression of H3K9ac and H4K5ac in the brain and spinal cord were detected by immunohistochemistry(IHC).Results Compared with the NC group,the maximum average clinical scores and the clinical scores of the 21th day of the model group was significantly increased(P<0.01).Compared with the model group,the maximum average clinical scores and the clinical scores of the 21th day of the TCM,DPCPX+TCM and hormone groups were significantly decreased(P<0.05).The results of WB showed that,compared with the NC group,A1AR in the brain and spinal cord of the model group was significantly decreased(P<0.01),while H3K9ac and H4K5ac were significantly increased(P<0.05).Compared with the model group,in DPCPX group A1AR in the brain was significantly decreased(P<0.05),H3K9ac and H4K5ac in the brain and spinal cord were significantly increased(P<0.05).In TCM group,A1AR in spinal cord was significantly increased(P<0.05),H3K9ac and H4K5ac in the brain was significantly decreased(P<0.05).In DPCPX+TCM group,A1AR in the brain and spinal cord was significantly increased(P<0.05),H4K5ac in the brain was significantly decrease

关 键 词：益肾达络饮 多发性硬化 实验性自身免疫性脑脊髓炎 腺苷A1受体 组蛋白乙酰化 

分 类 号：R285.5[医药卫生—中药学]