主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

JOURNAL OF BEIJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE ›› 2019, Vol. 42 ›› Issue (11): 923-933.doi: 10.3969/j.issn.1006-2157.2019.11.008

• Chinese Medicinal Pharmacology • Previous Articles     Next Articles

Influence of Siwu Tang on proliferation of MC3T3-E1 cells based on GPER-mediated EGFR/PI3K pathway*

Cui Lixia1, Shi Danning1, Jiao Shihong1, Zhao Piwen1#, Sun Liping1, Niu Jianzhao2   

  1. 1 School of Life Sciences, Beijing University of Chinese Medicine, Beijing 102488, China;
    2 School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 102488, China
  • Received:2019-05-06 Online:2019-11-30 Published:2019-12-03
  • Contact: Prof. Zhao Piwen, Ph.D., Doctoral supervisor. School of Life Sciences, Beijing University of Chinese Medicine, Intersection of Yangguang Nandajie Road and Baiyang Donglu Road, Fangshan District, Beijing 102488. E-mail: pwzhao@263.net

Abstract: Objective To investigate the influence of Siwu Tang (Four Ingredients Decoction) on proliferation efficacy of mouse embryonic osteoblasts (MC3T3-E1) based on G-protein-coupled estrogen receptor (GPER)-mediated pathway of epidermal growth factor receptor (EGFR)/ phosphatidylinositol 3-kinase (PI3K), and study the molecular mechanism. Methods Female SD rats (n=40) were randomly divided into normal group, estradiol group, and high-dose, mid-dose and low-dose Siwu Tang groups (each n=8). At the same time, the high-dose Siwu Tang group was divided into high-dose Siwu Tang+G1 group, high-dose Siwu Tang+G15 group, high-dose Siwu Tang+AG1478 group and high-dose Siwu Tang+LY294002 group. Taken E2 medicated serum as positive control, MC3T3-E1 was intervened with high-dose, mid-dose and low-dose Siwu Tang medicated serum. The influence of 24-h intervention with medicated serum on MC3T3-E1 proliferation was detected by using methyl thiazolyl tetrazolium test (MTT) in all groups. The influence of high-dose Siwu Tang in high dose on protein expressions of EGFR, PI3K, cyclin B1 and cyclin D1 in MC3T3-E1 were detected by suing Western blot assay and immunofluorescence technique(IF). The effective pathway and molecular mechanism of Siwu Tang in high dose influencing MC3T3-E1 proliferation were detected by using GPER agonist (G1), GPER antagonist (G15), EGFR inhibitor (AG1478) and PI3K inhibitor (LY294002). Results The proliferation of MC3T3-E1 was improved in high-dose, mid-dose and low-dose Siwu Tang groups compared with normal group (P<0.01), and G1 increased significantly the improving effect of Siwu Tang in high dose on proliferation of MC3T3-E1 (P<0.01). G15, AG1478 and LY294002 decreased the improving effect of Siwu Tang in high dose on proliferation of MC3T3-E1 (P<0.01). The Results of Western blotting assay and FIA showed that the protein expressions of EGFR, PI3K, cyclin B1 and cyclin D1 in MC3T3-E1 increased significantly in all Siwu Tang groups compared with normal group (P<0.01). G1 promoted the above-mentioned expressions (P<0.01), and G15, AG1478 and LY294002 reduced the above-mentioned expressions (P<0.01, P<0.05) compared with Siwu Tang in high dose. Conclusion Siwu Tang has significant improving effect on the proliferation of MC3T3-E1, and the effect mechanism may be related to the conduction of GPER-mediated EGFR/PI3K signaling pathway.

Key words: Siwu Tang (Four Ingredients Decoction), MC3T3-E1, estrogen, G-protein-coupled estrogen receptor, epidermal growth factor receptor

CLC Number: 

  • R285.5