摘要
目的:基于转录组学结合免疫浸润分析探讨红芪蜜炙前后配伍醋莪术抗结肠炎相关结直肠癌(CAC)的药效及潜在机制差异。方法:75只SPF级C57/6J小鼠随机分为5组:空白组、氧化偶氮甲烷(AOM)/葡聚糖硫酸钠(DSS)组、柳氮磺胺吡啶阳性对照组、红芪∶醋莪术(4∶1)组(6.825 g/kg)及炙红芪∶醋莪术(4∶1)组(6.825 g/kg)。采用AOM与DSS联合诱导CAC小鼠模型的同时,各给药组给药干预,空白组给予0.9%氯化钠溶液,干预17周后实验结束。计算各组小鼠体质量、DAI评分、结肠长度、肿瘤计数、生存指数、结肠长度/结肠质量、结肠及脾、胸腺、肝、肺、肾与心等脏器指数,HE染色观察小鼠结肠病变情况,ELISA法检测小鼠血清白介素(IL)-1β及肿瘤坏死因子(TNF)-α表达水平,有参转录组测序结合免疫浸润分析筛选并预测给药后免疫细胞表达情况,流式细胞术对所筛选的免疫细胞进行检测。结果:与AOM/DSS组比较,炙红芪-醋莪术与红芪-醋莪术均可显著升高CAC小鼠体质量、结肠长度、生存率及胸腺指数(P<0.05,P<0.01),显著降低DAI评分、肿瘤计数、HE病理评分、结肠长度/结肠质量及结肠、脾、胸腺、肝、肺等脏器指数与血清IL-1β与TNF-α表达水平(P<0.05,P<0.01);但从结肠HE病理切片评分及TNF-α水平来看,炙红芪-醋莪术的作用要显著优于红芪-醋莪术(P<0.05,P<0.01)。有参转录组测序结合免疫浸润分析发现,红芪-醋莪术与炙红芪-醋莪术均可通过上调小鼠脾脏CD4^(+)T淋巴细胞、CD8^(+)T淋巴细胞及CD19+B淋巴细胞表达量,降低CD4^(+)T/CD8^(+)T值;但对小鼠脾脏CD8^(+)T淋巴细胞表达的影响,炙红芪-醋莪术作用显著优于红芪-醋莪术(P<0.01)。结论:红芪-醋莪术与炙红芪-醋莪术均具防治CAC作用,机制与调节机体T、B淋巴细胞表达量,改善机体免疫,从而改善炎症状态,延缓炎症进程,改善结肠病变有关。但炙红芪-醋莪术防治CAC优于红芪-�
Objective:To study on efficacy and immunomodulatory mechanism for Hedysari Radix(HR)before and after honey-processing(HPHR)paired with vinegar-processed Curcumae Rhizoma(VPCR)against colitis-associated colorectal cancer(CAC).Methods:Seventy-five SPF-grade C57/6J mice were randomly divided into five groups:control group,Azoxymethane(AOM)/Dextran sulphate sodium salt(DSS)group,Sulfasalazine group,HR∶VPCR(4∶1)group(6.825 g/kg),and HPHR∶VPCR(4∶1)group(6.825 g/kg).While the CAC mouse model was induced by the combination of AOM and DSS.Each dosing group was given drug intervention and the control group was given saline,and the experiment was ended after 17 weeks of intervention.Body mass,DAI score,colon length,tumor counts,survival index,colon length/colon mass,colon and organ indices such as spleen,thymus,liver,lungs,kidneys and heart were calculated,and colonic lesions were observed by HE staining.The expression levels of interleukin(IL)-1βand tumor necrosis factor(TNF)-αin serum were detected by ELISA.The expression of the immune cells was screened and predicted by reference transcriptome sequencing combined with immune infiltration analysis.The screened immune cells were detected by flow cytometry.Results:Compared with AOM/DSS group,both HRVPCR and HPHR-VPCR significantly increased the body mass,colon length,survival rate and thymus index(P<0.05,P<0.01),and significantly decreased the DAI scores,tumor counts,HE pathology scores,colon length/colon mass,and indexes of organs such as the colon,spleen,thymus,liver,and lungs in association with the serum IL-1βand TNF-αexpression levels(P<0.05,P<0.01).However,the effect of HPHR-VPCR was significantly superior to HR-VPCR in terms of HE pathological scores in colon and TNF-αlevels in serum(P<0.05,P<0.01).The transcriptome sequencing combined with immune infiltration analysis revealed that both HR-VPCR and HPHR-VPCR could reduce the CD4^(+)T/CD8^(+)T values and up-regulated the expression of CD4^(+)T cells,CD8^(+)T cells,and CD19+B cells in spleen.However,for t
作者
张育贵
张转红
王彦钧
刘婷
高飞云
李越峰
ZHANG Yugui;ZHANG Zhuanhong;WANG Yanjun;LIU Ting;GAO Feiyun;LI Yuefeng(College of Pharmacy,Gansu University of Chinese Medicine,Lanzhou 730000,China;Scientific Research and Experimental Center,Gansu University of Chinese Medicine,Lanzhou 730000,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2024年第5期2349-2356,共8页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(No.81960713,No.82160750)
甘肃省教育厅产业支撑计划项目(No.2021CYZC-21)
甘肃省教育厅双一流重大科研项目(No.GSSYLXM-05)
甘肃省科学技术厅-科技计划(创新基地与人才计划)基础研究创新群体项目(No.21JR7RA569)
甘肃省中药炮制技术传承基地项目(No.甘卫中医函[2022]288号)
2024年省级人才项目(No.甘组通字[2024]4号)。
关键词
红芪-醋莪术
炙红芪-醋莪术
结肠炎相关结肠癌
药效
转录组学
免疫浸润
免疫调节
Hedysari Radix-vinegar-processed Curcumae Rhizoma
Honey-processed Hedysari Radix-vinegar-processed Curcumae Rhizoma
Colitis-associated colorectal cancer(CAC)
Efficacy
Transcriptomics
Immune infiltration
diilation
