基于转录组测序技术研究香砂六君子汤治疗慢性萎缩性胃炎的分子机制  

Molecular mechanism of Xiangsha Liujunzi Decoction in treating chronic atrophic gastritis based on transcriptome sequencing technology

作　　者：刘梦雅 梁永林[1] 成映霞 段永强 白敏 袁晓梅 刘自由 公艳霞 LIU Meng-ya;LIANG Yong-lin;CHENG Ying-xia;DUAN Yong-qiang;BAI Min;YUAN Xiao-mei;LIU Zi-you;GONG Yan-xia(Gansu University of Chinese Medicine,Lanzhou 730000,China;Gansu Province Laboratory Animal Industry Technology Center,Lanzhou 730000,China;Key Laboratory of Traditional Chinese Medicine for Prevention and Control of Regional High Incidence Diseases in Ningxia Ministry of Education,Ningxia Medical University,Yinchuan 750004,China;Afiliated Hospital of Traditional Chinese Medicine,Ningxia Medical University,Wuzhong 751100,China)

机构地区：[1]甘肃中医药大学,甘肃兰州730000 [2]甘肃省实验动物行业技术中心,甘肃兰州730000 [3]宁夏医科大学宁夏区域高发病中医药防治教育部重点实验室,宁夏银川750004 [4]宁夏医科大学附属中医医院,宁夏吴忠751100

出　　处：《中国中药杂志》2024年第18期4977-4985,共9页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金地区项目(81760830,82060828);国家中医药管理局重点学科建设项目(国中医药人教函[2023]85号);宁夏医科大学中医学院一流学科建设项目(ZY0019110305)。

摘　　要：基于转录组学技术探讨香砂六君子汤治疗慢性萎缩性胃炎(CAG)的分子机制并进行实验验证。通过1-甲基-3-硝基-1-亚硝基胍(MNNG)复合多因素法建立CAG大鼠模型,香砂六君子汤干预90 d。测定各组大鼠体质量和3 h进食量,采用HE染色观察各组大鼠胃组织病理改变,采用转录组学测序技术(RNA-Seq)测定各组大鼠胃组织中基因表达,对差异基因进行KEGG富集分析并对富集到的关键信号通路进行验证。结果显示,与空白组相比,模型组大鼠体质量、3 h进食量显著降低,胃组织表现典型的萎缩之征;与模型组相比,给药组大鼠体质量、3 h进食量显著增加,胃组织病理损伤均得到明显改善。RNA-Seq测序共获得香砂六君子汤改善CAG的相关基因944个,KEGG富集分析发现这些基因主要富集于多种涉及炎症、增殖、凋亡的信号转导途径以及环磷酸腺苷(cAMP)和环磷酸鸟苷(cGMP)/蛋白激酶G(PKG)通路。与空白组比较,模型组胃组织中腺苷A1受体(Adora1)、钙激活钾通道蛋白β2(Kcnmb2)mRNA表达显著升高,B型内皮素受体(Ednrb)、人Rho关联含卷曲螺旋蛋白激酶2(Rock2)、cGMP依赖性蛋白激酶Ⅱ(Prkg2)、环腺苷酸应答元件结合蛋白H抗体(Creb3l3)mRNA表达显著降低;与模型组比较,给药组胃组织中Adora1、Kcnmb2 mRNA表达显著降低,Ednrb、Rock2、Prkg2、Creb3l3 mRNA表达显著升高。与空白组比较,模型组胃组织和血浆中cAMP含量以及胃组织中Ras同源基因家族成员A(RhoA)、ROCK2、磷酸化肌球蛋白磷酸酶靶标亚基1(p-MYPT1)、磷酸化肌球蛋白轻链20(p-MLC20)蛋白表达均显著降低,胃组织和血浆中cGMP含量以及胃组织中PKG蛋白表达显著升高;与模型组比较,给药组胃组织和血浆中cGMP含量以及胃组织中PKG蛋白表达显著降低,胃组织中RhoA、ROCK2、p-MYPT1、p-MLC20蛋白表达显著升高。同时RT-qPCR结果表明cGMP/PKG通路关键基因的表达与RNA-Seq结果基本一致。综上所述,香Based on transcriptomics technology,this study investigated the molecular mechanisms of Xiangsha Liujunzi Decoction in treating chronic atrophic gastritis(CAG),which were confirmed through experimental validation.The CAG rat model was built by the MNNG composite multi-factor method,followed by a 90-day administration of Xiangsha Liujunzi Decoction.The study measured the rat body mass and 3-hour food intake in each group and observed the pathological changes in gastric tissue using HE staining.It used RNA sequencing(RNA-Seq)to determine the gene expression in the gastric tissue of rats in each group and then analyzed differential genes through KEGG enrichment analysis to validate enriched key signaling pathways.The results indicated that the body mass and 3-hour food intake in the model group were significantly lower than those in the blank group,alongside typical signs of gastric tissue atrophy.In contrast,the treatment group exhibited a significant increase in body mass and 3-hour food intake and significant improvements in the damage of gastric tissue pathology.A total of 944 genes associated with the improvement of CAG by Xiangsha Liujunzi Decoction were obtained via RNA-Seq.KEGG enrichment analysis revealed that these genes were mainly enriched in multiple signal transduction pathways involving signaling pathways such as inflammation,proliferation,apoptosis,as well as cyclic adenosine monophosphate(cAMP)and cyclic guanosine monophosphate(cGMP)/protein kinases(PKG)pathways.Compared to the blank group,the RNA-Seq results showed significant increases in adenosine A1 receptor recombina(Adora1)and BK channel subunit beta 3(Kcnmb2)mRNA expression in gastric tissue in the model group,while the expression of endothelin receptor type B gene(Ednrb),recombinant Rho associated coiled coil containing protein kinase 2(Rock2),cGMP-dependent protein kinase 2(Prkg2),and cAMP responsive element binding protein 3 like 3 gene(Creb3l3)mRNA was significantly decreased.Compared to the model group,the expression of Adora1 and Kcnmb2

关 键 词：香砂六君子汤 慢性萎缩性胃炎 cGMP/PKG通路 转录组测序技术 实验验证 

分 类 号：R285[医药卫生—中药学]