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
张丹 1张喜 1倪家慧 1杨良俊 2刘伟 3潘华峰1
作者信息
- 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,广州 510006;广州中医药大学第一附属医院,岭南医学研究中心,广州 510006
- 折叠
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-1 β、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.
Abstract
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-1 β and 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Key words
Gastric precancerous lesions(GPL)/Jianpi Huayu Jiedu Formula/Macrophage/Pyroptosis/Inflammation-carcinoma transition引用本文复制引用
基金项目
国家自然科学基金面上项目(81973816)
国家自然科学基金青年科学基金项目(82004300)
广东省重点领域研发计划项目(2020B1111100011)
广东省基础与应用基础研究基金项目(2023A1515011107)
广州市科学技术局基础研究计划(2023A04J1169)
省部共建中医湿证国家重点实验室开放课题面上项目(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(ZYYCXTD-C-202208)
出版年
2024
国家科技期刊平台
NSTL
万方数据