腺病毒介导的RNA干扰降低VEGF受体FLT-1,KDR转录抑制血管形成

本研究用细菌内同源重组的方法构建了针对VEGF受体FLT-1,KDR的siRNA表达重组腺病毒Ad H1-siRNA/FLT-1和Ad H1-siRNA/KDR,感染HUVEC细胞,观察两种病毒对HUVEC细胞体外增殖和形成微血管的干扰作用.通过RT-PCR实验表明,Ad H1-siRNA/FLT-1和Ad H1-siRNA/KDR均可特异性地下调血管内皮细胞(HUVEC)中的FLT-1mRNA和KDR mRNA水平.与对照组相比,FLT-1的mRNA水平下降为40%,KDR的mRNA水平下降为52%.Ad H1-siRNA/FLT-1和Ad H1-siRNA/KDR对HUVEC细胞增殖均有干扰作用,加入病毒9 d后,对照组的细胞平均计数为31.5×104/μL,Ad H1-siRNA/FLT-1干扰病毒对照组为28×104/μL,Ad H1-siRNA/KDR干扰组为21.5×104/μL.Ad H1-siRNA/FLT-1和Ad H1-siRNA/KDR对HUVEC细胞在Matrigel上形成微血管均有干扰作用,对照组每HPF形成微血管数量为10.75条,Ad H1-siRNA/FLT-1干扰病毒对照组为10.25条,实验组Ad H1-siRNA/KDR每HPF形成微血管数量为7条.证明Ad H1-siRNA/FLT-1,Ad H1-siRNA/KDR均可干扰血管形成.     

Effects of flavonoid derivatives on human microvascular endothelial cells

Some natural compounds, including flavonoids, are active in vasculature re-growth during hair follicle disruption, but their effects have not been yet evaluated directly on microvascular endothelial cells. Skin vascularisation regulates the physiological blood supply required for hair growth and its dysregulation is the basis of several human diseases. Follicle-derived vascular endothelial growth factor (VEGF) release from follicular keratinocytes promotes perifollicular vascularisation and increases follicle and hair size, while blockade of VEGF-mediated angiogenesis leads to impaired hair growth. Here, we tested three flavonoids, namely visnadin (VSD), hesperidin (HSP) and baicalin (BC), on cultured human microvascular endothelial cells (HMEC), comparing their effects with minoxidil (MXD), a synthetic drug broadly used in the treatment of androgenetic alopecia. The response to these compounds was assayed in terms of endothelial survival, proliferation, tubulogenesis and proangiogenic signalling. We show that BC promotes HMEC proliferation, while both VSD and MXD enhance tubulogenesis. Interestingly, only HSP increases VEGFR-2 phosphorylation.     

Ranking effects of candidate drugs on biological process by integrating network analysis and Gene Ontology

There are high preclinical attrition rates in current drug discovery. The efficient assessment approach in the high throughout candidate drugs screening still needs great improvement. We propose two hypotheses. First, both drug action process and biological process can be converted to a common space of gene or gene product profiling. Second, the strength of drug action on biological process can be realized in the context of biological network. Based on the above hypotheses, we establish an algorithm termed Network-based Assessment for Drug Action (NADA) to assess the action strength of candidate drugs on certain biological processes. Then NADA is used to prioritize the effects of six compounds from traditional Chinese medicine on endothelial cell migration, a simple process defined by Gene Ontology, in the biological network specific for a given pathological process, angiogenesis. The computational results are subsequently tested by the experiment on the migration of Human Umbilical Vein Endothelial Cells in vitro. The experimental ranks for six compounds generally agree with the predicted output of NADA. NADA also outperforms the DAVID and meet/min methods in terms of the experimental orders, suggesting that the network topological features may have a key role in catching the mechanistic relationship between drug action and biological process. Hopefully, the progress of network biology approaches for deciphering complex diseases will further expedite the preclinical screening and accelerate the development of treatment modalities.     

红景天苷上调大鼠心肌梗死后心肌组织VEGF的表达

目的:探讨红景天苷对大鼠心肌梗死后心肌组织血管内皮生长因子(VEGF)的表达调控作用及其可能机制。方法:建立大鼠心梗模型后,随机分为模型组、红景天苷3个不同剂量组,每组8只大鼠,另设假手术组8只。术后48 h,红景天苷组分别给予低、中、高,即10,20,40 mg·(kg·d)-1灌胃;模型组和假手术组给予生理盐水20 ml·(kg·d)-1灌胃。4周后处死大鼠,取大鼠心肌组织,反转录PCR(RT-PCR)法测定VEGF mRNA的表达。应用免疫组化和免疫印迹法分析左心室心肌组织VEGF蛋白的表达情况。结果:RT-PCR结果表明,和模型组相比,红景天苷各剂量组VEGF mRNA的表达均明显升高(P<0.01)。免疫组化和免疫印迹分析结果表明,和模型组相比,红景天苷各剂量组心肌组织胞浆中VEGF蛋白的表达均显著升高(P<0.01)。结论:红景天苷通过上调VEGF的表达而促大鼠心肌梗死后心肌组织的血管新生。     

Targeting Bone Metastases with a Bispecific Anticancer and Antiangiogenic Polymer–Alendronate–Taxane Conjugate

A polymer therapeutic designed for combination anticancer and antiangiogenic therapy inhibited the proliferation of prostate carcinoma cells and the proliferation, migration, and tube‐formation of endothelial cells. The nanoconjugate was formed from an N‐(2‐hydroxypropyl)methacrylamide (HPMA) copolymer, the bisphosphonate alendronate (for bone targeting), and the chemotherapy agent paclitaxel (PTX), which is cleaved by cathepsin B (see scheme).

     

Natural Phenolic Acid,Product of the Honey Bee,for the Control of Oxidative Stress,Peritoneal Angiogenesis,and Tumor Growth in Mice

Tumor-associated macrophages (TAM) are key regulators of the link between inflammation and cancer, and the interplay between TAM and tumor cells represents a promising target of future therapeutic approaches. We investigated the effect of gallic acid (GA) and caffeic acid (CA) as strong antioxidant and anti-inflammatory agents on tumor growth, angiogenesis, macrophage polarization, and oxidative stress on the angiogenic model caused by the intraperitoneal (ip) inoculation of Ehrlich ascites tumor (EAT) cells (2.5 × 10⁶) in Swiss albino mouse. Treatment with GA or CA at a dose of 40 mg/kg and 80 mg/kg ip was started in exponential tumor growth phase on days 5, 7, 9, and 11. On day 13, the ascites volume and the total number and differential count of the cells present in the peritoneal cavity, the functional activity of macrophages, and the antioxidant and anti-angiogenic parameters were determined. The results show that phenolic acids inhibit the processes of angiogenesis and tumor growth, leading to the increased survival of EAT-bearing mice, through the protection of the tumoricidal efficacy of M1 macrophages and inhibition of proangiogenic factors, particularly VEGF, metalloproteinases -2 and -9, and cyclooxygenase-2 activity.     

Antiangiogenic Activity and in Silico Cereblon Binding Analysis of Novel Thalidomide Analogs

Due to its antiangiogenic and anti-immunomodulatory activity, thalidomide continues to be of clinical interest despite its teratogenic actions, and efforts to synthesize safer, clinically active thalidomide analogs are continually underway. In this study, a cohort of 27 chemically diverse thalidomide analogs was evaluated for antiangiogenic activity in an ex vivo rat aorta ring assay. The protein cereblon has been identified as the target for thalidomide, and in silico pharmacophore analysis and molecular docking with a crystal structure of human cereblon were used to investigate the cereblon binding abilities of the thalidomide analogs. The results suggest that not all antiangiogenic thalidomide analogs can bind cereblon, and multiple targets and mechanisms of action may be involved.     

5-Formylhonokiol exerts anti-angiogenesis activity via inactivating the ERK signaling pathway

Our previous report has demonstrated that 5-formylhonokiol (FH), a derivative of honokiol (HK), exerts more potent anti-proliferative activities than honokiol in several tumor cell lines. In present study, we first explored the antiangiogenic activities of 5-formylhonokiol on proliferation, migration and tube formation of human umbilical vein endothelial cells (HUVECs) for the first time in vitro. Then we investigated the in vivo antiangiogenic effect of 5-formylhonokiol on zebrafish angiogenesis model. In order to clarify the underlying molecular mechanism of 5-formylhonokiol, we investigated the signaling pathway involved in controlling the angiogenesis process by western blotting assay. Wound-healing results showed that 5-formylhonokiol significantly and dose-dependently inhibited migration of cultured human umbilical vein enthothelial cells. The invasiveness of HUVEC cells was also effectively suppressed at a low concentration of 5-formylhonokiol in the transwell assay. Further F-actin imaging revealed that inhibitory effect of 5-formylhonokiol on invasion may partly contribute to the disruption of assembling stress fiber. Tube formation assay, which is associated with endothelial cells migration, further confirmed the anti-angiogenesis effect of 5-formylhonokiol. In in vivo zebrafish angiogenesis model, we found that 5-formylhonokiol dose-dependently inhibited angiogenesis. Furthermore, western blotting showed that 5-formylhonokiol significantly down-regulated extracellular signal-regulated kinase (ERK) expression and inhibited the phosphorylation of ERK but not affecting the total protein kinase B (Akt) expression and related phosphorylation, suggesting that 5-formylhonokiol might exert anti-angiogenesis capacity via down-regulation of the ERK signal pathway. Taken together, these data suggested that 5-formylhonokiol might be a viable drug candidate in antiangiogenesis and anticancer therapies.