基于TGF-β1/TSC22D1介导的CAMs通路探讨艾灸改善乳腺癌化疗性粒缺的机制

前期发现，艾灸神阙、足三里和三阴交，能一定程度上改善乳腺癌患者化疗性中性粒细胞减少症，与上调外周血中性粒细胞TSC22D1表达有关；RNA-seq结合生物信息学分析，TGFβ1、TSC22D1为关键分子，细胞粘附分子（CAMs）通路尤为重要。本研究在前期研究基础上，采用乳腺癌环磷酰胺化疗小鼠，研究TGFβ1/TSC22D1介导的CAMs通路在艾灸升粒机制的核心调节作用：应用流式细胞、磁珠吸附等现代分子生物学技术与方法，①拟明确艾灸对乳腺癌环磷酰胺化疗小鼠骨髓、外周血和脾脏不同来源中性粒细胞绝对值的变化规律；②初步阐明艾灸调节骨髓、外周血和脾脏不同来源中性粒细胞TGFβ1/TSC22D1表达，以及对ITGB1/3、JAM3、CLDN5、ESAM等粘附分子的作用机制；③探索不同时间灸程对艾灸升粒效应机制的影响，以期提高艾灸防治乳腺癌化疗性粒缺的临床疗效，为艾灸升粒的临床推广和应用提供科学依据。

The previous studies have shown that the mechanism of moxibustion on Shenque, Zusanli and Sanyinjiao acupoints which can improve chemotherapy-induced neutropenia to a certain extent, is related to the up-regulation of TSC22D1 expression in peripheral neutrophils of breast cancer patients. Combined with bioinformatics analysis, RNA-seq found that TGFβ1 and TSC22D1 are the key molecules, among them cell adhesion molecules (CAMs) pathway is particularly important. Based on previous research results, we are going to study on the critical regulatory effect of CAMs pathway mediated by TGFβ1/TSC22D1 in the mechanism of moxibustion increasing neutrophils using cyclophosphamide treated mice with breast cancer .With modern molecular biological techniques and methods such as flow cytometry and magnetic bead adsorption, etc., our objectives are: ① to clarify the variation of moxibustion on absolute neutrophil counts from different sources including bone marrow, peripheral blood and spleen in cyclophosphamide treated mice with breast cancer; ②to preliminarily clarify the mechanism of moxibustion regulating the expression of TGFβ1/TSC22D1 in neutrophils from different sources including bone marrow, peripheral blood and spleen, as well as ITGB1/3、JAM3、CLDN5、ESAM and other adhesion molecules; ③ to explore the effect of different moxibustion course on the mechanism of increasing neutrophils, so as to improve the clinical effect of moxibustion on prevention and treatment of chemotherapy-induced neutropenia in breast cancer, as well as providing scientific basis for clinical application of moxibustion.