Elucidation the binding mechanism of Nelumbo nucifera-derived isoquinoline alkaloids as Rho-kinase 1 inhibitors by molecular docking and dynamic simulation.

Rho-kinase 1 (ROCK1) is a key molecular target for controlling smooth muscle (SM) contraction in asthma, gastrointestinal disorders, hypertension. Embryos of lotus seed (Nelumbo nucifera) are traditional folk herbs widely used in treating various diseases which are closely related to SM contraction. With the aim of explaining the mechanism of embryos of lotus seed, 27 isoquinoline alkaloids were isolated from the embryos of lotus seed, the inhibitory activity of these alkaloids against ROCK1 were virtual screened via molecular docking and molecular dynamics (MD) simulations. The docking results indicated that 5 bisbenzylisoquinolines (BBIs) and 1 tribenzylisoquinoline (TBI) were potent inhibitors with high binding affinity for both A and B chains of ROCK1 (AcRock and BcRock). The MD results also revealed that neoliensinine (28) was the most potent inhibitor, which was corresponding to the irreversible relaxation effect of neoliensinine on SM. Moreover, through the MD simulation, it also indicated that neoliensinine (28) interacted in its stretched conformation through polar solvation interactions and van der Waal forces. Finally, with the best calculation results, the inhibition effect of neoliensinine (28) on the contraction of vascular smooth muscle cells (VSMCs) and ROCK1 was also confirmed by several biological tests.Communicated by Ramaswamy H. Sarma.

Expression of Fibroblast Growth Factor 4 in a Rat Model of Polydactyly of the Thumb Induced by Cytarabine.

BACKGROUND The aim of this study was to assess the expression and mechanisms of fibroblast growth factor 4 in polydactyly of the thumb induced by cytarabine. MATERIAL AND METHODS Rats were intraperitoneally injected with cytarabine at different gestation periods (12.5 days, 13.5 days, and 14.5 days) to establish a polydactyly of the thumb model. Then, the expression of FGF4 in polydactyly was studied by whole-mount in situ hybridization. We used hematoxylin & eosin stain and cartilage stain to investigate the development of the skeleton and tissues in the embryo. Pictures were taken to determine the general shape of the deformity, then X-rays were taken to detect bone distortion of the rats born with a congenital malformation. RESULTS In the experimental group (11.5 days, 12.5 days, 13.5 days, and 14.5 days), whole-mount in situ hybridization showed that the FGF4 expression at the tip of the embryonic limb bud was significantly increased compared with the control group and FGF4 was distributed in a wider range and lasted longer than in the control group (P<0.01). HE staining and cartilage staining showed that there was an extra metacarpal bone and a phalanx in the rats with polydactyly of the thumb (P<0.01). Images of the deformed limbs showed polydactyly and syndactyly of the thumb in the rats. Further X-ray examination revealed 1 extra metacarpal bone and 1 extra phalanx. CONCLUSIONS Cytarabine can induce polydactyly and syndactyly of the thumb in rats. In this process, cytarabine can induce the expression of FGF4 on the tip of the embryonic limb bud, which further leads to abnormal development of the embryonic limb bud and eventually causes a congenital deformity.