[Frontiers In Bioscience, Landmark, 24, 1050-1059, March 1, 2019]

C1q contributes to post-stroke angiogenesis via LAIR1-HIF1α-VEGF pathway

Guofeng Fan1, Qiming Li2, Jian Qian1

1Department of Emergency Medicine, Nanjing Drum Tower Hospital Affiliated to Nanjing University School of Medicine, Nanjing, 210008, P.R. China, 2Department of Emergency Medicine, the First Affiliated Hospital of Shenzhen University, Health Science Center, the Second People’s Hospital of Shenzhen, Shenzhen, 518035, P.R. China

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Materials and Methods
3.1. Experimental animals
3.2. Western blot analysis
3.3. Measurement of vascular density
3.4. Cell culture
3.5. Hypoxia culture condition
3.6. Cell viability assay
3.7. VEGF production detection
3.8. Migration assay
3.9. Tube Formation
3.10. RNA interference
3.11. Measurement of brain water content
3.12. Statistical Analysis
4. Results
4.1. Increased C1q expression in IBZ enhances angiogenesis in rat with pMCAO
4.2. C1q promotes viability and angiogenic function of RBMECs and HBMECs
4.3. C1q upregulates VEGF expression and secretion in RBMECs, HBMECs and in IBZ in pMCAO rats
4.4. C1q activates LAIR1-HIF1-VEGF axis in RBMECs
4.5. Administration of C1q enhances neovasculariation after pMCAO in rat
5. Discussion
6. Acknowledgement
7. References

1. ABSTRACT

Vascular remodeling is a critical event following a stroke. It is a well known fact that C1q is the first molecule in the complement classical pathway. However, its role in the neovascularization that ocurs after a stroke, remains unclear. In this study, we investigated the effects of C1q on post-stroke angiogeneis in pMCAO rats. We found that increased C1q levels in IBZ enhanced angiogenesis in rats with pMCAO. C1q promoted viability and angiogenic function of RBMECs and HBMECs. Upregualtion of VEGF expression and secretion by C1q was also observed in RBMECs, HBMECs and in IBZ in pMCAO rats. Furthermore, we demonstrated that C1q enhanced angiogenic function of RBMECs through its receptor, LAIR1. The results show that administration of C1q enhanced neovascularization and reduced brain edema after pMCAO in rats. On the basis of these findings, we suggest that C1q plays an important role in post-stroke angiogenesis at least through LAIR- HIF1α-VEGF axis. C1q shows promise as a potential therapeutic candidate for stroke treatment.

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Abbreviations: HBMECs: Human Brain Microvascular Endothelial Cells; RBMECs : Rat Brain Microvascular Endothelial Cells; HIF1α: Hypoxia Inducible Factor 1 α; IBZ: Ischemic Boundary Zone; LAIR1: Leukocyte-associated Inhibitory Receptor-1; pMCAO: Permanent Middle Cerebral Artery Occlusion; VEGF: Vascular Endothelial Growth Factor; PVDF: Polyvinylidene Difluoride

Key Words: C1q, Stroke, Angiogenesis, LAIR1, VEGF, HIF1 alpha

Send correspondence to: Jian Qian, Department of Emergency Medicine, the Affiliated Drum Tower Hospital of Nanjing University School of Medicine, Nanjing, 210008, P.R. China, Tel: 86-25-0831096666, Fax: 86-25-86508960 E-mail: jianqian08@gmail.com