清肠温中方通过调节CD4^(+)T细胞HIF-1α对溃疡性结肠炎相关性结直肠癌小鼠瓦博格效应的影响  

Qingchang Wenzhong decoction modulates the HIF-1αin CD4^(+)T cells to influence the Warburg effect in mice with ulcerative colitis-associated colorectal cancer

作　　者：章晓思 李军祥[2] 胡俊聪 杨安琪 傅智浩 张黎明[1] 石磊[2] 毛堂友 王志斌[2] ZHANG Xiaosi;LI Junxiang;HU Juncong;YANG Anqi;FU Zhihao;ZHANG Liming;SHI Lei;MAO Tangyou;WANG Zhibin(Beijing University of Chinese Medicine,Beijing,100029,China;Department of Gastroenterology,Dongfang Hospital,Beijing University of Chinese Medicine)

机构地区：[1]北京中医药大学,北京100029 [2]北京中医药大学东方医院脾胃肝胆科

出　　处：《中国中西医结合消化杂志》2025年第3期310-316,共7页Chinese Journal of Integrated Traditional and Western Medicine on Digestion

基　　金：中央高校基本科研业务费专项资金项目(No:2022-JYB-JBZR-008);国家中医药管理局高水平中医药重点学科中西医结合临床(消化病学)(No:zyyzdxk-2023271)。

摘　　要：目的:观察清肠温中方对溃疡性结肠炎相关性结直肠癌(ulcerative colitis-associated colorectal cancer,UC-CRC)小鼠的整体干预效应,分析清肠温中方调节CD4^(+)T细胞中HIF-1α对UC-CRC小鼠瓦博格效应的影响。方法:将48只C57BL/6雄性小鼠随机分为正常组、模型组、阳性药物组及清肠温中方高、中、低剂量组,每组8只。采用诱变剂氧化偶氮甲烷和致炎剂葡聚糖硫酸钠诱导UC-CRC模型。自第2周注射氧化偶氮甲烷开始,阳性药物组每天予以美沙拉嗪0.04 g/kg剂量灌胃,清肠温中方高、中、低剂量组每天分别予以18.50、9.25、4.63 g/kg的剂量灌胃,连续10周。每周测量1次小鼠体重,检测小鼠血清中乳酸含量、结直肠的长度及瘤体数目;苏木精-伊红染色法观察小鼠结直肠组织形态学改变;酶联免疫吸附法检测小鼠结直肠组织中IL-6含量;RT-qPCR法检测小鼠结直肠组织中HIF-lα、Glut1、HK2、PKM2及LDHA的mRNA表达;免疫荧光双染检测CD4^(+)T细胞中HIF-1α的表达。结果:与正常组比较,模型组小鼠体重下降,结直肠长度明显缩短(P<0.0001),瘤体数目明显增多(P<0.0001);血清中乳酸含量显著升高(P<0.01);黏膜上皮欠完整,腺体结构紊乱或消失,隐窝萎缩,核大而深染,肿瘤组织形成;促炎因子IL-6含量明显升高(P<0.0001);结直肠组织中HIF-1α(P<0.0001)、Glut1(P<0.0001)、HK2(P<0.0001)、PKM2(P<0.01)及LDHA(P<0.01)显著升高,CD4^(+)T细胞中HIF-1α明显升高。经清肠温中方干预后小鼠体重下降情况得到改善,结直肠长度明显延长(P<0.0001),瘤体数目显著减少(P<0.001),血清中乳酸含量显著下降(P<0.05),可见上皮细胞形态较完整,有效改善了隐窝结构,减少肿瘤组织的形成;促炎因子IL-6含量明显下降(P<0.0001);结直肠组织中HIF-1α(P<0.001)、G1ut1(P<0.0001)、HK2(P<0.0001)、PKM2(P<0.05)及LDHA(P<0.01)显著下降,CD4^(+)T细胞中HIF-1α显著降低。结论:清肠温中方一定程度上可以减少UC-CRC模�Objective To observe the overall intervention effect of Qingchang Wenzhong decoction(QCWZD)on mice of colitis-associated colorectal cancer(UC-CRC)and analyze the impact of QCWZD on the Warburg effect in CD4^(+)T cells via regulating HIF-1α.Methods C57BL/6 male mice were randomly divided into a control group,model group,positive drug group,a high-dose QCWZD group,medium-dose QCWZD group,and low-dose QCWZD group,with 8 mice in each group.The model of UC-CRC was induced by the mutagenic agent azoxymethane(AOM)and the inflammatory agent dextran sulfate sodium(DSS).Since the second week of AOM,the positive drug group has been administered mesalazine at a dose of 0.04 g/kg by gavage daily.In contrast,the high,medi um,and low-dose groups of QCWZD have been administered with doses of 18.50 g/kg,9.25 g/kg,and 4.63 g/kg by gavage daily,respectively,for a consecutive period of 10 weeks.The body weight of the mice was measured once a week;serum lactic acid content was detected;the length of the colorectal tract and the number of tumors were recorded;histomorphological changes in the colorectal tissue were observed using Hematoxylin and Eosin staining;the levels of interleukin(IL)-6 in the colorectal tissue were measured by enzyme-linked immunosorbent assay(ELISA);the mRNA expression of HIF-lα,Glut1,HK2,PKM2,and LDHA in the colorectal tissue was detected by real-time fluorescence quantitative PCR(RT-qPCR);and the expression of HIF-1αin CD4^(+)T cells was assessed by immunofluorescence double staining.Results Compared with the control group,the model group showed a significant reduction in colorectal length(P<0.0001)and a marked increase in the number of tumors(P<0.0001);serum lactic acid content significantly increased(P<0.01);the mucosal epithelium was incomplete with disordered or absent glandular structures,crypt atrophy,enlarged and hyperchromatic nuclei,and tumor tissue formation;the levels of pro-inflammatory factors IL-6 significantly increased(P<0.0001);the expression of HIF-1α(P<0.0001),Glut1(P<0.0001),HK2(P<0.00

关 键 词：清肠温中方 溃疡性结肠炎相关性结直肠癌 缺氧诱导因子1Α CD4^(+)T细胞 瓦博格效应 

分 类 号：R256.3[医药卫生—中医内科学]