Combination of ferulic acid, ligustrazine and tetrahydropalmatine inhibits invasion and metastasis through MMP/TIMP signaling in endometriosis

Yi Tan; Chengling Zhang; Ying Zhang; Xueshan Dai; Qinghua Wei; Jiahui Wei; Pingli Xu; Yi Chen

doi:10.7717/peerj.11664

Combination of ferulic acid, ligustrazine and tetrahydropalmatine inhibits invasion and metastasis through MMP/TIMP signaling in endometriosis

PeerJ. 2021 Jun 28:9:e11664. doi: 10.7717/peerj.11664. eCollection 2021.

Authors

Yi Tan^#^{1

2

3

4}, Chengling Zhang^#^{1

2

3

4}, Ying Zhang^{1

2

3

4}, Xueshan Dai^{1

2

3

4}, Qinghua Wei^{1

2

3

4}, Jiahui Wei^{1

2

3

4}, Pingli Xu⁵, Yi Chen^{1

2

3

4}

Affiliations

¹ College of Pharmaceutical Sciences & Chinese Medicine, Southwest University, Chongqing, China.
² Chongqing Key Laboratory of New Drug Screening from Traditional Chinese Medicine, Chongqing, China.
³ Pharmacology of Chinese Materia Medica-the Key Discipline Constructed by the State Administration of Traditional Chinese Medicine, Chongqing, China.
⁴ National Demonstration Center for Experimental Pharmacy Education (Southwest University), Chongqing, China.
⁵ Chongqing Hospital of Traditional Chinese Medicine, Chongqing, China.

^# Contributed equally.

Abstract

Background: The design of the combination of ferulic acid, ligustrazine and tetrahydropalmatine (FLT) is inspired by the Chinese herbal prescription Foshou San. Previous work has shown that FLT inhibited endometriosis growth in rat autograft models. However, the mechanism behind this is unclear. MMP/TIMP signaling is considered as the vital pathway of metastasis and invasion in endometriosis. In this study, we aim to disclose effects of FLT on MMP/TIMP signaling in invasion and metastasis during endometrial cells and xenograft endometriosis.

Methods: In vivo, effect of FLT on endometriosis was evaluated in a xenogeneic mice model. In vitro, cell viability assay was performed with an IC50 measurement of FLT in hEM15A and HEC1-B cells. The effect of FLT on invasion and metastasis was analyzed in scratch wound and transwell assay. Gene and protein expression of MMP/TIMP signaling were detected by qPCR and Western blotting.

Results: In xenograft endometriosis, FLT reduced ectopic volume without effect on weight. FLT inhibitory effects on cell growth exhibited a dose-dependent manner in hEM15A and HEC1-B cells. IC50s of FLT in hEM15A cells were 839.30 ± 121.11 or 483.53 ±156.91 μg·ml^-1 after the treatment for 24 or 48 h, respectively. In HEC1-B cells, IC50 values of 24 or 48 h were 625.20 ± 59.52 or 250.30 ± 68.12 μg·ml^-1. In addition, FLT significantly inhibited invasion and metastasis in scratch wound and transwell assay. Furthermore, FLT inactivated MMP/TIMP signaling with decreasing expression of MMP-2/9, and an enhancing expression of TIMP-1.

Conclusions: MMP/TIMP inactivation is a reasonable explanation for the inhibition of FLT on invasion and metastasis in endometriosis. This result reveals a potential mechanism on the role of FLT in endometriosis and may benefit for its further application.

Keywords: Endometriosis; Ferulic acid; Invasion and metastasis; Ligustrazine; MMP/TIMP signaling; Tetrahydropalmatine.

Grants and funding

This work was supported by the National Natural Science Foundation of China (No. 81773984, 81402441), the Chongqing Natural Science Foundation of China (No. CSTC2020JCYJ-MSXM1639), and the Traditional Chinese Medicine Research Project of Chongqing Health Bureau (No. 2020ZY023665). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.