摘要
目的:探讨艾灸通过转化生长因子β1(TGFβ1)/转化生长因子β诱导基因22结构域家族蛋白1(TSC22D1)信号通路干预乳腺癌化疗性中性粒细胞减少症的作用机制.方法:以4T1乳腺癌细胞接种于Balb/c小鼠第3对乳腺脂肪垫下构建乳腺癌小鼠模型,以环磷酰胺(CTX)溶液腹腔注射乳腺癌小鼠构建乳腺癌化疗性中性粒细胞减少症(CIN)小鼠模型.分为正常对照组、荷瘤对照组、化疗模型组、艾灸组和西药组,每组4只.艾灸组取"神阙""足三里""三阴交"穴进行艾灸,西药组予以人粒细胞集落刺激因子10 μg/kg皮下注射.干预结束后采用HE染色观察骨髓组织病理变化,取眼眶静脉血采用磁珠吸附法提取外周血中性粒细胞,流式细胞术鉴定提取纯度,采用ELISA法检测小鼠中性粒细胞TGFβ1、TSC22D1、JAM3、ESAM、ICAM-1、VCAM-1蛋白含量,RT-qPCR法和Western Blot法检测TGFβ1、TSC22D1、JAM3、ESAM、ICAM-1、VCAM-1 mRNA和蛋白表达.结果:与荷瘤对照组比较,化疗模型组TGFβ1、JAM3 mRNA和蛋白表达显著升高(P＜0.05),TSC22D1、ESAM、ICAM-1、VCAM-1 mRNA和蛋白表达显著降低(P＜0.05);与化疗模型组比较,艾灸组、西药组TGF β1、JAM3 mRNA和蛋白表达显著降低(P＜0.05),艾灸组TSC22D1、ESAM、ICAM-1、VCAM-1 mRNA和蛋白表达显著升高(P＜0.05).结论:艾灸通过干预TGFβ1/TSC22D1信号通路并调控JAM3、ESAM、ICAM-1、VCAM-1黏附分子的表达,增加中性粒细胞黏附功能,可能是艾灸改善乳腺癌化疗性中性粒细胞减少症的机制之一.
Abstract
Objective:To observe the mechanism of moxibustion on improving chemotherapy-induced neutropenia(CIN)in breast cancer through transforming growth factor beta 1(TGF β1)/transforming growth factor beta stimulated clone-22 domain family protein-1(TSC22D1)signaling pathway.Methods:4T1 breast cancer cells were inoculated under the third pair of breast pads in Balb/c mice to establish a mouse model of breast cancer.Breast cancer mice were subjected to intraperitoneal injection of cyclophosphamide(CTX)solution to develop a breast cancer CIN mouse model.The mice were divided into normal control group,tumor-bearing control group,chemotherapy model group,moxibustion group,and western medicine group,with 4 mice in each group.In the moxibustion group,Shenque,Zusanli,and Sanyinjiao were used for moxibustion.The western medicine group received a subcutaneous injection of G-CSF 10 μg/kg.After the intervention,hematoxylin-eosin(HE)staining was used to observe pathological changes in the bone marrow tissue.Peripheral blood neutrophils were extracted from the orbital vein blood using a magnetic bead adsorption method,and flow cytometry was used to identify extraction purity.The ELISA method was applied to detect the protein levels of TGFβ 1,TSC22D1,JAM3,ESAM,ICAM-1,and VCAM-1 in the mice's neutrophils,while RT-qPCR and Western Blot methods were used to examine the mRNA and protein expressions of TGF β 1,TSC22D1,JAM3,ESAM,ICAM-1,and VCAM-1.Results:Compared with tumor control group,the mRNA and protein expressions of TGF β 1 and JAM3 of neutrophils in chemotherapy group were increased(P＜0.05),while the mRNA and protein expressions of TSC22D1,ESAM,ICAM-1,VCAM-1 of neutrophils in chemotherapy group were decreased(P＜0.05).Compared with the chemotherapy model group,the mRNA and protein expressions of TGF β1 and JAM3 of neutrophils in the moxibustion group and western medicine group were decreased(P＜0.05),while the mRNA and protein expressions of TSC22D1,ESAM,ICAM-1 and VCAM-1 of neutrophil in moxibustion group were increased(P＜0.05).Conclusion:Moxibustion could increase the adhesion function of neutrophils through the TGFβ1/TSC22D1 signaling pathway,induces neutrophils from infantile to mature,and then improves CIN of breast cancer.
基金项目
国家自然科学基金项目(82004446)
上海市针灸临床医学研究中心(20MC1920500)
促进市级医院临床技能与临床创新能力三年行动计划(SHDC2020CR4057)
上海市科学技术委员会科技计划项目(22Y11923000)
上海市科学技术委员会科技计划项目(20Y21901100)
国家中医药领军人才"岐黄学者"()
国家科技期刊平台
NSTL
万方数据