北京中医药大学学报2018，Vol.41Issue(3)：196-202,7.DOI:10.3969/j.issn.1006-2157.2018.03.004
回阳生肌方不同拆方对巨噬细胞表型转化的调节作用
Regulation effects of different disassembled formulas of Huiyang Shengji Fang on macrophage phenotype transition
摘要
Abstract
Objective To observe the influence of Huiyang Shengji Fang(Restoring Yang and Promoting Tissue Regeneration Decoction, HYSJF) and its decomposed formulas-Yiqi Wenyang Fang (Qi-supplementing and Yang-warming Decoction,YQWYF) and Huoxue Tongluo Fang(Blood-activating and collateral-freeing Decoction,HXTLF) on macrophage phenotype inversion,and discuss the mechanism of YQWYF and HXTLF in regulating macrophage phenotype inversion during the period of chronic skin ulcer. Methods THP-1 cells were stimulated and transformed into macrophages with PMA. After stimulated with LPS and INF-γ for 48 h, the macrophages were transformed into M1 macrophages. Lovastatin (10 μmol /L) was taken as control drug. The influence of YQWYF and HXTLF on activities of macrophages was observed by using CCK-8 method, and their influence on the phagocytosis of macrophages was detected by using neutral red uptake assay (NRU). The expressions of iNOS mRNA and Arg-1 mRNA were detected by using polymerase chain reaction (PCR) after M1 macrophages stimulated with lovastatin,YQWYF and HXTLF for 24 h. The levels of supernate TNF-α,IL-6,IL-12, IL-10 and VEGF were detected by using CBA method. The level of supernate TGF-β was detected by using ELISA. Results YQWYF,in dose of 4 mg/L (low-dose YQWYF group),0.8 mg/L (mid-dose YQWYF group) and 0.16 mg/L (high-dose YQWYF group), and HXTLF, in dose of 32 mg/L (low-dose HXTLF group),6.4 mg/L (mid-dose HXTLF group) and 1.28 mg/L (high-dose HXTLF group) had no influence on proliferation of macrophages but improved phagocytosis of NRU of macrophages. The expression of iNOS mRNA of M1 macrophages increased significantly after stimulated by LPS and INF-γ, and decreased significantly in lovastatin group, YQWYF group and HXTLF group compared with model group (P <0.01). The expression of Arg-1 mRNA of M2 macrophages decreased significantly after stimulated by LPS and INF-γ compared with normal group, was improved significantly in lovastatin group, YQWYF group and HXTLF group compared with model group (P <0. 01), and decreased significantly in YQWYF group and HXTLF group compared with lovastatin group. After M1 macrophages stimulated by above drugs for 24 h, the levels of TNF-α, IL-6 and IL-1 decreased significantly, and levels of VEGF and TGF-β increased significantly. The level of IL-10 increased significantly in high-dose YQWYF group (P<0.05),The levels of IL-10,VEGF and TGF-β had statistical difference in HXTLF group and high-dose YQWYF group compared with lovastatin group. Conclusion YQWYF and HXTLF can inhibit expression of iNOS mRNA of M1 macrophages and induce expression of Arg-1 mRNA of M2 macrophages, and can inhibit the secretion of M1 macrophage cytokine and improve secretion of M2 macrophage cytokine.关键词
回阳生肌方/慢性疮面愈合/益气温阳/活血通络/巨噬细胞表型/人单核细胞株Key words
Huiyang Shengji Fang(Restoring Yang and Promoting Tissue Regeneation Deloction)/supplementing qi and warming yang/activating blood and freeing collateral vessels/macrophages pheno-type/human monocytic line分类
医药卫生引用本文复制引用
林燕,何秀娟,赵京霞,薛妍,蒙玉娇,李萍..回阳生肌方不同拆方对巨噬细胞表型转化的调节作用[J].北京中医药大学学报,2018,41(3):196-202,7.基金项目
国家自然科学基金项目(No.81403409,No.81403270,No.81774328)National Natural Science Foundation of China(No.81403409,No.81403270,No.81774328) （No.81403409,No.81403270,No.81774328）
