Inhibition of eNOS by L-NAME resulting in rat hind limb developmental defects through PFKFB3 mediated angiogenetic pathway

Abstract

l-arginine/NOS/NO signaling pathway plays a critical role in controlling variety of vascular diseases. However, whether NOS inhibition by L-NAME suppresses late embryonic development is undefined. The aim of this study is to determine whether NOS inhibition by L-NAME is critical for late embryonic rat hind limb development. The pregnant rat at E13.5 administrated L-NAME by consecutive intraperitoneal injection. The embryos been harvested from E16.5 to E 20.5. Hematoxylin and Eosin Staining, Immunofluorescence and Immunohistochemistry performed to determine hind limb Vasculogenesis, HUVEC culture, Adenoviral PFKFB3 infection, Real time PCR and western blot were performed to determine whether l-arginine/NOS/NO pathway controlling late embryonic hind limb development through PFKFB3 mediated angiogenetic pathway. NOS inhibition by L-NAME resulting in late embryonic hind limb developmental defects characterized by severe hemorrhage. The in vivo studies showed that NOS inhibition strongly suppressed hind limb angiogenetic remodeling by impairing differentiation of endothelial cells and smooth muscle cells, and extracellular matrix synthesis. For underlie mechanism, our studies indicated that L-NAME treatment dramatically suppresses PFKFB3 expression in hematopoietic progenitor cells, tubulogenetic endothelial cells and smooth muscle cells. Knockdown of PFKFB3 dramatically inhibits the expression of angiogenetic genes, as well as tubulogenesis and extracellular matrix related genes. Taken together, our data in this study demonstrated that l-arginine-eNOS-NO pathway is important for rat hind limb development during late embryonic stage. This could be both a useful animal model and a promising therapeutic treatment for defects of late embryonic developmental hind limbs.

Restoration of Mitochondrial Function Is Essential in the Endothelium-Dependent Vasodilation Induced by Acacetin in Hypertensive Rats

“…Furthermore, as shown in Figure 3 B, a left shift in the dose-response curve with acacetin exhibiting a greater endothelium-dependent vessel relaxation was detected in WKY rats. Therefore, the above results suggested that acacetin improved vasodilation through activating the NO mediated pathway [ 17 , 18 ].…”

Impact of Non-pharmacological Chronic Hypertension on a Transgenic Rat Model of Cerebral Amyloid Angiopathy

“…For example, it was reported that AD-like symptoms in Tg-SwDI mice worsened with pharmacologically induced chronic HTN ( Kruyer et al, 2015 ). In this study, the endothelial nitric oxide synthase (eNOS) inhibitor, L-NAME ( Wu et al, 2020 ) was used. In similar studies, HTN was pharmacologically induced by eNOS inhibition in conjunction with administration of angiotensin II in the Tg2576 mouse model of AD-like pathology ( Passos et al, 2016 ; Nyúl-Tóth et al, 2020 ).…”

Wenshenyang recipe treats infertility through hormonal regulation and inflammatory responses revealed by transcriptome analysis and network pharmacology

“…NO is an important free radical species that is important in embryonic development. The activity of NO and NOS is necessary for embryos to initiate the formation of Frontiers in Pharmacology frontiersin.org the vasculature, and inhibition of NOS can lead to developmental defects or death of the embryo (Wu et al, 2020;Hitchler and Domann, 2021). Nuclear receptors play a very important role in the regulation of the final stage of ovarian follicle growth.…”