Yao Molly, Ganguly Samayita, Shin Jane Hae Soo, Elbayoumi Tamer
Department of Pharmaceutical Sciences, College of Pharmacy-Glendale, Midwestern University, Cholla Hall 216, 19555 N. 59th Ave., Glendale, AZ 85308, USA.
College of Graduate Studies, Midwestern University, Science Hall, 19555 N. 59th Ave., Glendale, AZ 85308, USA.
Methods Protoc. 2021 Oct 14;4(4):74. doi: 10.3390/mps4040074.
Homeostasis of vascular tone is intricately and delicately maintained systemically and locally, by autonomic nerves and hormones in the blood and by intimal vasoactive substances, respectively. The balance can be acutely or chronically interrupted secondary to many alterations, especially under pathological conditions. Excessive matricellular glycoprotein thrombospondin 1 (TSP1) levels in circulation have been found to play an important role in ischemia-reperfusion injuries of different organs, by acutely suppressing vasorelaxation and chronically remodeling vascular bed. Our laboratory has been interested in identifying new drug moieties, which can selectively and effectively counteract TSP1-induced vascular dysfunction, in order to address associated clinical complications. Preliminary studies using computational docking and molecular models revealed potential drug candidates for further evaluation via vascular functional bioassay to prove the antagonism using an ex vivo vascular model. Herein, we described an efficient screening method for the identification of active drug candidates, by adapting a multiwire myograph system to perform a protocol with different treatments, in the presence of pathological levels of TSP1. We discussed the promising pharmacological evaluation results and suggested suitable modification for versatile applications. We also described the necessity of pre-determination of optimal resting tension to obtain the maximal response, if the experimental test model is different from those with determined optimal resting tension.
血管张力的稳态通过血液中的自主神经和激素以及内膜血管活性物质分别在全身和局部进行复杂而精细的维持。这种平衡可能会因许多改变而急性或慢性中断,尤其是在病理条件下。已发现循环中过多的基质细胞糖蛋白血小板反应蛋白1(TSP1)水平在不同器官的缺血再灌注损伤中起重要作用,它通过急性抑制血管舒张和慢性重塑血管床来实现。我们实验室一直致力于鉴定新的药物部分,这些药物部分可以选择性且有效地对抗TSP1诱导的血管功能障碍,以解决相关的临床并发症。使用计算对接和分子模型的初步研究揭示了潜在的药物候选物,可通过血管功能生物测定进一步评估,以使用离体血管模型证明其拮抗作用。在此,我们描述了一种有效的筛选方法,通过采用多线肌张力测定系统在存在病理水平的TSP1时进行不同处理的方案来鉴定活性药物候选物。我们讨论了有前景的药理学评估结果,并建议进行适当修改以用于多种应用。我们还描述了如果实验测试模型与已确定最佳静息张力的模型不同,则预先确定最佳静息张力以获得最大反应的必要性。