文献类型：期刊文献

中文题名：半夏泻心汤含药肠吸收液对胃癌微环境中PMN-MDSCs凋亡的影响

英文题名：Effect of Banxia Xiexintang-containing Intestinal Absorption Solution on PMN-MDSCs Apoptosis in Gastric Cancer Microenvironment

作者：魏晶晶[1];朱中博[1];刘喜平[1];李沛清[1];陈启明[2];戴丽蓉[1];施丽娟[1];段海婧[1];王庆苗[1]

第一作者：魏晶晶

机构：[1]甘肃中医药大学甘肃省中药新产品创制工程实验室,甘肃省中医方药挖掘与创新转化重点实验室,兰州730000;[2]兰州大学第一医院,兰州730013

第一机构：甘肃中医药大学科研实验中心（甘肃省中医药标准化技术委员会秘书处）

年份：2023

卷号：29

期号：10

起止页码：58

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2023_2024】；

基金：国家自然科学基金项目(81860813,81860782);甘肃省高等学校创新基金项目(2020B-007);甘肃省委组织部陇原青年创新创业人才项目(2020RCXM183);甘肃省中医方药挖掘与创新转化重点实验室开放基金项目(ZYFY-2020-003)。

语种：中文

中文关键词：胃癌微环境;髓源性抑制细胞;细胞凋亡;半夏泻心汤

外文关键词：gastric cancer microenvironment;myeloid-derived suppressor cells;apoptosis;Banxia Xiexintang

摘要：目的:观察半夏泻心汤含药肠吸收液对胃癌微环境中多形核髓系来源抑制细胞(PMN-MDSCs)凋亡的影响。方法:制备半夏泻心汤含药肠吸收液,以Transwell小室将胃癌细胞与PMN-MDSCs非接触共培养建立胃癌微环境模型,采用细胞增殖与活性检测-8(CCK-8)法筛选0.63 g·m L^(-1)复溶液制备而成的0~100%半夏泻心汤含药肠吸收液的最佳干预浓度及时间,分为空白组、模型组、奥沙利铂组(10 mg·L^(-1))及半夏泻心汤组(26%、18%、10%含药肠吸收液),采用CCK-8法检测PMN-MDSCs的增殖,流式细胞术检测PMN-MDSCs的凋亡,蛋白免疫印迹法(Western blot)检测PMN-MDSCs胃癌微环境凋亡相关蛋白,B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、胱天蛋白酶-3(Caspase-3)蛋白表达水平。结果:作用24、48 h,与空白组比较,5%、50%、75%、100%半夏泻心汤含药肠吸收液对PMN-MDSCs的抑制率均有升高((P<0.05,P<0.01),作用72 h,除50%半夏泻心汤含药肠吸收液对PMN-MDSCs的抑制率低于48 h(P<0.01),其余浓度各自抑制率差异无统计学意义,且48 h半数抑制浓度(IC_(50))为18.40%,说明18%半夏泻心汤含药肠吸收液在48 h为最佳干预浓度及时间。与空白组比较,模型组PMN-MDSCs的存活率明显升高(P<0.05),凋亡率明显下降(P<0.05);与模型组比较,半夏泻心汤组PMN-MDSCs存活率均明显降低(P<0.05),凋亡率明显升高(P<0.05),以半夏泻心汤组(26%含药肠吸收液)最为明显(P<0.05)。与空白组比较,模型组PMN-MDSCs促凋亡蛋白Bax、Caspase-3表达明显降低(P<0.05),抗凋亡蛋白Bcl-2表达明显上升(P<0.05)。与模型组比较,奥沙利铂组、半夏泻心汤组PMN-MDSCs Caspase-3蛋白表达升高(P<0.05),Bcl-2蛋白表达明显降低(P<0.05),奥沙利铂组、半夏泻心汤组(10%含药肠吸收液)Bax蛋白表达升高(P<0.05)。结论:半夏泻心汤含药肠吸收液能通过调控胃癌微环境PMN-MDSCs凋亡相关蛋白Bax、Caspase-3及Bcl-2的表达,诱导PMN-MDSCs凋亡。
Objective:To observe the effect of Banxia Xiexintang(BXT)-containing intestinal absorption solution on the apoptosis of polymorphonuclear myeloid-derived suppressor cells(PMN-MDSCs)in gastric cancer microenvironment.Method:BXT-containing intestinal absorption solution was prepared,and gastric cancer cells and PMN-MDSCs were non-contact co-cultured in Transwell chamber to establish gastric cancer microenvironment.Cell counting kit-8(CCK-8)assay was used to screen the optimal intervention concentration and time of 0-100%BXT-containing intestinal absorption solution prepared by 0.63 g·mL^(-1) reconstitution solution.Cells were classified into blank group,model group,oxaliplatin group(10 mg·L^(-1)),and BXT(26%,18%,10%BXT-containing intestinal absorption solution)group,and the apoptosis of PMN-MDSCs was detected by flow cytometry.The expression of apoptosis-related B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),and cysteine-aspartic acid protease-3(Caspase-3)in PMN-MDSCs was detected by Western blot.Result:After treatment for 24 h and 48 h,the PMN-MDSCs-inhibiting rate was increased by 5%,50%,75%,and 100%BXT-containing intestinal absorption solution compared with that in the blank group(P<0.05,P<0.01).At 72 h,the PMN-MDSCs-inhibiting rate by 50%BXT-containing intestinal absorption solution was lower than that at 48 h(P<0.01),and the PMN-MDSCs-inhibiting rate by 5%,75%,and 100%BXT-containing intestinal absorption solution showed no significant difference from that at 48 h.Moreover,the half-maximal inhibitory concentration(IC_(50))at 48 h was 18.40%.Thus,18%BXT-containing intestinal absorption solution and 48 h were the optimal intervention concentration and time.The survival rate of PMN-MDSCs in model group was higher than that in the blank group(P<0.05),and the apoptosis rate was lower than that in the blank group(P<0.05).Compared with model group,BXT containing intestinal absorption solution lowered the survival rate and raised apoptosis rate of PMN-MDSCs(P<0.05),particularly the 26%BXT-containing intestinal absorption solution(P<0.05).The expression of Bax and Caspase-3 in PMN-MDSCs was lower in the model group than in the blank group(P<0.05),and the expression of Bcl-2 was higher in the model group than in the blank group(P<0.05).The expression of Caspase-3 in PMN-MDSCs increased(P<0.05)and the expression of Bcl-2 decreased(P<0.05)in oxaliplatin group and BXT group compared with those in the model group.The expression of Bax rose in oxaliplatin group and BXT group(10%BXT-containing intestinal absorption solution)(P<0.05).Conclusion:BXT can induce the apoptosis of PMN-MDSCs by regulating the expression of apoptosis-related proteins Bax,Caspase-3,and Bcl-2 in gastric cancer microenvironment.