基于转录组学探讨四神丸治疗慢性溃疡性结肠炎的作用机制  

Study on the mechanism of Sishen Pills in treating chronic ulcerative colitis based on transcriptomics

作　　者：李晓玲 吴玉泓 柳春[1] 李海龙[1] 殷银霞 李宬钰 尚立宇 梁永林[1] LI Xiaoling;WU Yuhong;LIU Chun;LI Hailong;YIN Yinxia;LI Chengyu;SHANG Liyu;LIANG Yonglin(Gansu University of Chinese Medicine,Lanzhou 730000,China;South China Hospital Affiliated to Shenzhen University.Shenzhen 518111,China;Shenzhen Hospital of Beijing University of Chinese Medicine(Longgang),Shenzhen 51811l,China)

机构地区：[1]甘肃中医药大学,兰州730000 [2]深圳大学附属华南医院,深圳518111 [3]北京中医药大学深圳医院(龙岗),深圳518111

出　　处：《中华中医药杂志》2024年第5期2504-2509,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81860810);2022年度甘肃省中医药科研课题(No.GZKP-2022-36);2022年度中医学一级学科“岐黄英才”导师专项基金博导项目(No.2022-5);2023年甘肃中医药大学(博士)研究生创新创业基金项目(No.2023CX16)。

摘　　要：目的:基于转录组学分析四神丸治疗溃疡性结肠炎(UC)的机制。方法:108只SPF级雄性C57BL/6小鼠随机选出18只为空白组,其余90只采用病证结合造模方法建立脾肾阳虚型UC模型。脾肾阳虚型UC模型建立后,将其随机分为模型组、四神丸高、中、低剂量组和阳性对照美沙拉秦组,每组18只,K组与M组给予蒸馏水灌胃,各治疗组给予对应药物及剂量灌胃治疗,连续21 d。观察各组小鼠一般状态,HE染色观察各组小鼠结肠黏膜组织病理变化;运用转录组学测序技术检测空白组、模型组、四神丸高剂量组小鼠结肠组织的基因表达谱,并将显著差异表达基因进行基因功能注释(GO)、京都基因与基因组百科全书(KEGG)富集分析;运用Western Blot和RT-qPCR分别检测小鼠结肠组织中瞬时受体电位V1、M8(TRPV1、TRPM8)蛋白及mRNA的表达。结果:与空白组比较,模型组小鼠一般状态差,结肠组织见严重病理损伤;与模型组比较,各治疗组小鼠一般状态、结肠组织损伤情况均得到不同程度的改善,其中四神丸各剂量组中以高剂量组改善最为明显。转录组学分析结果显示,空白组vs.模型组和模型组vs.高剂量组差异基因广泛参与了味觉转导、嗅觉传导、炎症介质对TRP通道的调节及细胞因子-细胞因子受体相互作用等相关信号通路,主要参与对温度刺激的反应的生物过程。Western Blot和RTqPCR验证结果显示,与空白组比较,模型组小鼠结肠组织TRPV1蛋白及mRNA表达水平显著降低(P<0.01),TRPM8蛋白及mRNA表达水平显著升高(P<0.01);与模型组比较,高剂量组小鼠结肠组织TRPV1蛋白及mRNA表达水平显著升高(P<0.01),TRPM8蛋白及mRNA表达水平显著降低(P<0.01)。结论:四神丸可能通过调控TRP通道相关蛋白及基因的表达,修复受损结肠黏膜组织,达到治疗UC的目的。Objective:To analyze the mechanism of Sishen Pills in the treatment of ulcerative colitis(UC)based on transcriptomics.Methods:Eighteen of 108 SPF male C57BL/6 mice were randomly selected as blank group(group K),and the other 90 mice were established with spleen and kidney yang deficiency UC model by disease and syndrome modeling method.After the establishment of the UC model of spleen-kidney yang deficiency,they were randomly divided into model group(group M),Sishen Pills high-dose,medium-dose and low-dose groups(groups G,Z and D)and Mesalazine positive control group(group Y),with 18 individuals/group.Groups K and M were given distilled water by intragastric administration,and each treatment group was given corresponding drugs and doses by intragastric administration for 21 consecutive days.The general state of mice in each group was observed,and the pathological changes of colonic mucosa were observed by HE staining.Transcriptomic sequencing technology was used to detect the gene expression profiles of colon tissues of mice in groups K,M and G,and the significantly differentially expressed genes were annotated into gene ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis;Western Blot and RT-qPCR were used to detect the expression of transient receptor potential vanilloid 1(TRPV1)and transient receptor potential melastatin 8(TRPM8)proteins and genes in mouse colon tissues,respectively.Results:Compared with group K,the general state of mice in group M was poor,and the colon tissue was severely damaged.Compared with group M,the general state and colon tissue damage of mice in groups G,Z,D and Y were improved to varying degrees,among which group G,Z and D,group G showed the most obvious improvement.Transcriptomic analysis showed that the K vs.M and M vs.G differential genes were widely involved in taste transduction,olfactory transduction,the regulation of TRP channels by inflammatory mediators and the interaction between cytokines and cytokine receptors,and were mainly involved in biologic

关 键 词：四神丸 溃疡性结肠炎 脾肾阳虚 转录组学 TRP通道 

分 类 号：R285.5[医药卫生—中药学]