健脾化瘀解毒方抑制巨噬细胞焦亡治疗胃癌前病变的机制  

Mechanism of Jianpi Huayu Jiedu Formula ameliorating gastric precancerous lesions through inhibiting macrophage pyroptosis

作　　者：张丹[1] 张喜 倪家慧 杨良俊 刘伟[1,3] 潘华峰 ZHANG Dan;ZHANG Xi;NI Jiahui;YANG Liangjun;LIU Wei;PAN Huafeng(Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Tongde Hospital of Zhejiang Province,Hangzhou 310012,China;Lingnan Medical Research Center,the First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510006,China)

机构地区：[1]广州中医药大学,广州510006 [2]浙江省立同德医院,杭州310012 [3]广州中医药大学第一附属医院,岭南医学研究中心,广州510006

出　　处：《中华中医药杂志》2024年第8期3964-3970,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(No.81973816);国家自然科学基金青年科学基金项目(No.82004300);广东省重点领域研发计划项目(No.2020B1111100011);广东省基础与应用基础研究基金项目(No.2023A1515011107);广州市科学技术局基础研究计划(No.2023A04J1169);省部共建中医湿证国家重点实验室开放课题面上项目(No.SZ2022KF17);国家中医药管理局中医药创新团队及人才支持计划项目(No.ZYYCXTD-C-202208)。

摘　　要：目的:探讨健脾化瘀解毒方通过抑制巨噬细胞焦亡治疗胃癌前病变(GPL)的机制。方法:空白组小鼠常规饲养,造模组予以N-甲基-N-亚硝基脲(MNU)自由饮用隔日禁食法构建GPL模型,随机分为模型组,健脾化瘀解毒方高、低剂量组和维酶素组,每组12只,分别进行干预后,HE染色、AB-PAS染色观察小鼠胃黏膜组织病理学变化,Western Blot、RT-qPCR检测细胞焦亡相关指标、免疫荧光染色观察CDX2、Ki67、F4/80表达情况及F4/80与NLRP3共定位情况。体外构建J774A.1焦亡模型,予健脾化瘀解毒方含药血清干预,观察细胞焦亡相关分子表达情况。结果:与空白组比较,GPL小鼠胃黏膜NLRP3、Caspase-1、GSDMD-N、CDX2、Ki67、F4/80表达显著升高(P<0.01),NLRP3与F4/80存在共定位现象,集中分布于黏膜层。经健脾化瘀解毒方干预后,上述分子表达下降(P<0.01,P<0.05),且NLRP3与F4/80共定位现象散在分布。体外实验中,与空白组比较,模型组NLRP3、GSDMD-N、IL-1β、IL-18表达升高(P<0.01),PI染色阳性细胞比例增加(P<0.01),健脾化瘀解毒方可减少上述分子表达(P<0.01,P<0.05),减少PI染色阳性细胞比例(P<0.01,P<0.05)。结论:健脾化瘀解毒方可通过抑制巨噬细胞焦亡治疗GPL。Objective:To explore the mechanism of Jianpi Huayu Jiedu Formula(JPHYJD)ameliorating gastric precancerous lesions(GPL)through inhibiting macrophage pyroptosis.Methods:Mice in control group were fed regularly and others were exposed to MNU and were fed every other day to construct GPL models and were divided into model group,JPHYJD high-dose group,JPHYJD low-dose group and Vitacoenzyme(VIT)group randomly,with 10 in each group.HE staining and AB-PAS staining were used to evaluate the effect of JPHYJD.The expression of pyroptosis-related molecules were observed through Western Blot and RT-qPCR.CDX2,Ki67,F4/80 and the colocalization of F4/80 and NLRP3 were checked through immunofluorescent staining.J774A.1 cells were exposed to LPS and ATP to construct pyroptosis models and then JPHYJD-contained serum.Results:Compared with the control group,the expression of NLRP3,Caspase-1,GSDMD-N,CDX2,Ki67 and F4/80 in model group increased(P<0.01).The colocalization of F4/80 and NLRP3 were observed and concentrated in the mucosal layer.After treated by JPHYJD,molecules above decreased(P<0.01,P<0.05)and the colocalization only scattered.In vitro,compared with the control group,there showed a higher level of NLRP3,GSDMD-N,IL-1βand IL-18(P<0.01),and more PI-positive cells in model group(P<0.01),which were decreased by JPHYJD(P<0.01,P<0.05).Conclusion:JPHYJD could ameliorate GPL through inhibiting macrophage pyroptosis.

关 键 词：胃癌前病变 健脾化瘀解毒方 巨噬细胞 细胞焦亡 炎-癌转化 

分 类 号：R285.5[医药卫生—中药学]