摘要
OBJECTIVE: To observe the effects of serum containing Mahuang(Herba Ephedra Sinica) or Wuweizi(Fructus Schisandrae Chinensis) on the migration of alveolar macrophages(AM) and interstitial macrophages(IM) from normal rats, and to analyze and compare the mechanisms leading to cell migration differences.METHODS: Rats were randomly divided into three groups: Mahuang(Herba Ephedra Sinica), Wuweizi(Fructus Schisandrae Chinensis), and blank serum. After treatment with the herbs, serum was extracted from the rats. AM and IM were isolated from normal rats and cultured. The effects of Mahuang(Herba Ephedra Sinica) and Wuweizi(Fructus Schisandrae Chinensis) medicated serum on normal rat AM and IM chemotactic migration were determined by transwell assays. The CC chemokine receptor(CCR) 2, CCR5, voltage-gated Kvl. 3 K^+channel(Kv1. 3), and voltage-gated Kvl. 5 K^+channel(Kv1. 5) protein levels were analyzed by western blotting.RESULTS: The migration quantities of AM and IM in the Mahuang(Herba Ephedra Sinica) and Wuweizi(Fructus Schisandrae Chinensis) medicated serum groups were significantly higher than those in the blank serum group(P < 0.01). Compared with the Wuweizi(Fructus Schisandrae Chinensis) medicated serum group, the migration quantity of cultured rat AM in the Mahuang(Herba Ephedra Sinica) medicated serum group was significantly increased(P <0.01). Meanwhile, compared with the Mahuang(Herba Ephedra Sinica) medicated serum group, the migration quantity of cultured rat IM in the Wuweizi(Fructus Schisandrae Chinensis) medicated serum group was significantly increased(P < 0.01).CCR2, CCR5, Kv1. 3, and Kv1. 5 proteins were expressed on the AM cell surface, and showed significantly higher expression in the Mahuang(Herba Ephedra Sinica) medicated serum group compared with the Wuweizi(Fructus Schisandrae Chinensis)medicated serum group. In contrast, CCR5, Kv1.3,and Kv1.5 proteins were expressed on the IM cell surface, and showed significantly higher expression in the Wuweizi(Fructus Schisandrae Chinensis)medicated serum group compared with the Mahuang(Herba Ephedra Sinica) medicated serum group.CONCLUSION: Mahuang(Herba Ephedra Sinica)and Wuweizi(Fructus Schisandrae Chinensis) can promote AM and IM migration ability, with Mahuang(Herba Ephedra Sinica) targeting AM more apparently and Wuweizi(Fructus Schisandrae Chinensis) targeting IM more apparently. The mechanism may be that, by stimulating cells, Mahuang(Herba Ephedra Sinica) and Wuweizi(Fructus Schisandrae Chinensis) promote expression of CCR2 and CCR5 receptors on the AM and IM cell surface,which pass signals to Kvl.3 and Kvl.5 ion channels,leading to changes in the cytoskeleton, and ultimately promoting chemotactic cell migration.
OBJECTIVE： To observe the effects of serum containing Mahuang（Herba Ephedra Sinica） or Wuweizi（Fructus Schisandrae Chinensis） on the migration of alveolar macrophages（AM） and interstitial macrophages（IM） from normal rats, and to analyze and compare the mechanisms leading to cell migration differences.METHODS： Rats were randomly divided into three groups： Mahuang（Herba Ephedra Sinica）, Wuweizi（Fructus Schisandrae Chinensis）, and blank serum. After treatment with the herbs, serum was extracted from the rats. AM and IM were isolated from normal rats and cultured. The effects of Mahuang（Herba Ephedra Sinica） and Wuweizi（Fructus Schisandrae Chinensis） medicated serum on normal rat AM and IM chemotactic migration were determined by transwell assays. The CC chemokine receptor（CCR） 2, CCR5, voltage-gated Kvl. 3 K~＋channel（Kv1. 3）, and voltage-gated Kvl. 5 K~＋channel（Kv1. 5） protein levels were analyzed by western blotting.RESULTS： The migration quantities of AM and IM in the Mahuang（Herba Ephedra Sinica） and Wuweizi（Fructus Schisandrae Chinensis） medicated serum groups were significantly higher than those in the blank serum group（P 〈 0.01）. Compared with the Wuweizi（Fructus Schisandrae Chinensis） medicated serum group, the migration quantity of cultured rat AM in the Mahuang（Herba Ephedra Sinica） medicated serum group was significantly increased（P 〈0.01）. Meanwhile, compared with the Mahuang（Herba Ephedra Sinica） medicated serum group, the migration quantity of cultured rat IM in the Wuweizi（Fructus Schisandrae Chinensis） medicated serum group was significantly increased（P 〈 0.01）.CCR2, CCR5, Kv1. 3, and Kv1. 5 proteins were expressed on the AM cell surface, and showed significantly higher expression in the Mahuang（Herba Ephedra Sinica） medicated serum group compared with the Wuweizi（Fructus Schisandrae Chinensis）medicated serum group. In contrast, CCR5, Kv1.3,and Kv1.5 proteins were expressed on the IM cell surface, and showed significantly higher expression in the Wuweizi（Fructus Schisandrae Chinensis）medicated serum group compared with the Mahuang（Herba Ephedra Sinica） medicated serum group.CONCLUSION： Mahuang（Herba Ephedra Sinica）and Wuweizi（Fructus Schisandrae Chinensis） can promote AM and IM migration ability, with Mahuang（Herba Ephedra Sinica） targeting AM more apparently and Wuweizi（Fructus Schisandrae Chinensis） targeting IM more apparently. The mechanism may be that, by stimulating cells, Mahuang（Herba Ephedra Sinica） and Wuweizi（Fructus Schisandrae Chinensis） promote expression of CCR2 and CCR5 receptors on the AM and IM cell surface,which pass signals to Kvl.3 and Kvl.5 ion channels,leading to changes in the cytoskeleton, and ultimately promoting chemotactic cell migration.
基金
Supported by The National Natural Science Foundation of China(No.81373887)
The Traditional Chinese Medicine Standardization Project of the State Administration of Traditional Chinese Medicine(No.ZYXX-2014)
The Beijing Municipal Administration of Traditional Chinese Medicine"Generation 3+3"Project-Jin Shiyuan the Well-Known Old Man Chinese Medicine Studio Station(Beijing University of Chinese Medicine)Construction Project
