The influence of Tongqiao Huashuan Decoction on the expression of VEGF and EphB2 in MCAO model rats.

BACKGROUND: To observe the effect of Miao medicine and Tongqiao Huashuan Decoction on the expression of vascular endothelial growth factor (VEGF) and ephrin-B2 (EphB2) in the frontal lobe of the involved side and the cerebellum of rat models of middle cerebral artery occlusion (MCAO) and to reveal the pharmacological mechanism of Tongqiao Huashuan Decoction in treating acute ischemic stroke. METHODS: Seventy healthy male SD rats were randomly divided into sham operation, model, salvia miltiorrhiza (S. miltiorrhiza), and Miao medicine groups. Modified Longa's method was used to prepare a cerebral ischemia reperfusion model. After the operation, the rats in the sham operation group and model group were intragastrically administered with saline, those in the Miao medicine group were intragastrically administered with Tongqiao Huashuan Decoction, and those in the S. miltiorrhiza group were intraperitoneally injected with S. miltiorrhiza. After 14 days of administration, the neurological deficit scores of the rats in each group were compared before and after treatment. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of the brain tissues in the right infarcted areas of the rats. VEGF expression in the frontal lobe and cerebellum was observed through immunohistochemistry, and in situ hybridization to detect EphB2 expression in the frontal lobe and cerebellum. RESULTS: The neurological deficit scores were significantly improved in the Miao medicine and S. miltiorrhiza groups aftertreatment compared with those of the model group (P<0.05) and was higher in the Miao medicine group than in the S. miltiorrhiza group. The VEGF expression in the right frontal lobe and cerebellum was significantly increased in the Miao medicine and S. miltiorrhiza groups (P<0.05) with the former having higher levels than the latter (P<0.05). EphB2 expression was significantly increased in the frontal lobe and cerebellum in the Miao medicine and S. miltiorrhiza groups (P<0.05) and was higher in the frontal lobe of the Miao medicine group than that of the S. miltiorrhiza group (P<0.05) but was not significantly different in the cerebellum in the S. miltiorrhiza and Miao medicine groups (P>0.05). CONCLUSIONS: Tongqiao Huashuan Decoction can improve the neurological function score and promote the VEGF expression in the frontal lobe and cerebellum and the EphB2 expression in the frontal lobe of the involved side of MCAO rats. The pharmacological mechanism of Tongqiao Huashuan Decoction in treating acute ischemic stroke may be related to its regulation of VEDF and EphB2 expression in the distal part of the involved side.

Synergistic Effect of TPD7 and Berberine against Leukemia Jurkat Cell Growth through Regulating Ephrin-B2 Signaling.

TPD7, a novel biphenyl urea taspine derivative, and berberine have presented inhibition on VEGFR2 that can be regulated by ephrin-B2 reverse signaling through interactions with the PDZ domain. The purpose of this study is to investigate the inhibitory effect of the combination of TPD7 and berberine (TAB) on T-cell acute lymphoblastic leukemia cell growth. TPD7 and berberine together synergistically inhibited the proliferation of Jurkat cells. Also, the combination of TAB induced G1 -phase cell-cycle arrest by downregulating the level of cyclin D1, cyclin E, and CDC2. Furthermore, the combination of TAB significantly enhanced apoptosis in Jurkat cells, and the apoptosis most likely resulted from the modulation of the level of Bcl-2 family members. Most importantly, the concomitant treatment simultaneously regulated the ephrin-B2 and VEGFR2 signaling, as well as modulated the MEK/ERK and PTEN/PI3K/AKT/mTOR signaling. Therefore, the combination treatment of TAB may be a promising therapeutic method in treating T-cell acute lymphoblastic leukemia. Copyright © 2017 John Wiley & Sons, Ltd.

Berberine inhibits the proliferation and migration of breast cancer ZR-75-30 cells by targeting Ephrin-B2.

BACKGROUND: Berberine, a plant-derived compound isolated from Coptis chinensis used in traditional Chinese medicine, has been shown to possess anti-cancer properties. However, no study has shown that berberine could target ephrin-B2, which plays a critical role in cell proliferation and migration. PURPOSE: The aim of this study is to investigate the effect of berberine on cancer cell growth and migration, through the regulation of ephrin-B2 and downstream signaling molecules. METHODS: In this study, a high ephrin-B2-expressing cell membrane chromatography method was developed to investigate 48 crude extracts from traditional Chinese medicine that could act on ephrin-B2. Cell proliferative and wound-healing assays were used to study the effect of berberine on cancer cell growth and migration. The mechanism of berberine was investigated using western blot. RESULTS: Berberine was isolated from C. chinensis extracts and showed activity on the HEK293/ephrin-B2 cell membrane chromatography column. Berberine showed a greater inhibitory effect in high-expressing ephrin-B2 cells (HEK293/ephrin-B2 cells) than in normal HEK293 cells, and decreased the expression of ephrin-B2 and its PDZ binding proteins, which indicates that ephrin-B2 is a target of berberine. Furthermore, berberine downregulates the phosphorylation of VEGFR2 and downstream signaling members (AKT and Erk1/2), which in turn downregulates the expression of MMP2 and MMP9. CONCLUSION: The above data confirm the inhibitory effects of berberine on ZR-75-30 cell proliferation and cell migration. Overall, our studies demonstrate that berberine inhibits cell growth and migration by targeting ephrin-B2.

Cartilage-specific deletion of ephrin-B2 in mice results in early developmental defects and an osteoarthritis-like phenotype during aging in vivo.

BACKGROUND: Ephrins and their related receptors have been implicated in some developmental events. We have demonstrated that ephrin-B2 (EFNB2) could play a role in knee joint pathology associated with osteoarthritis (OA). Here, we delineate the in vivo role of EFNB2 in musculoskeletal growth, development, and in OA using a cartilage-specific EFNB2 knockout (EFNB2(Col2)KO) mouse model. METHODS: EFNB2(Col2)KO was generated with Col2a1-Cre transgenic mice. The skeletal development was evaluated using macroscopy, immunohistochemistry, histomorphometry, radiology, densitometry, and micro-computed tomography. Analyses were performed at P0 (birth) and on postnatal days P15, P21, and on 8-week- and 1-year-old mice. RESULTS: EFNB2(Col2)KO mice exhibited significant reduction in size, weight, length, and in long bones. At P0, the growth plates of EFNB2(Col2)KO mice displayed increased type X collagen, disorganized hyphertrophic zone, and decreased mineralization. At P15, mutant mice demonstrated a significant reduction in VEGF and TRAP at the chondro-osseous junction and a delay in the secondary ossification, including a decrease in bone volume and trabecular thickness. At P21 and 8 weeks old, EFNB2(Col2)KO mice exhibited reduced bone mineral density in the total skeleton, femur and spine. One-year-old EFNB2(Col2)KO mice demonstrated OA phenotypic features in both the knee and hip. By P15, 27 % of the EFNB2(Col2)KO mice developed a hip locomotor phenotype, which further experiments demonstrated reflected the neurological midline abnormality involving the corticospinal tract. CONCLUSION: This in vivo study demonstrated, for the first time, that EFNB2 is essential for normal long bone growth and development and its absence leads to a knee and hip OA phenotype in aged mice.