RESUMEN
OBJECTIVE: To investigate the effect of biflavonoid 4'-O-methylochnaflavone (MF) on palmitic acid-induced endothelial dysfunction in rat cavernous endothelial cells (RCECs). METHODS: The isolated RCECs were commercially available and randomly divided into four groups: normal+BSA group (NC group), palmitic acid (PA) group, MF group, and icariside â ¡ (ICA â ¡) group. The protein expression levels of protein kinase B (PKB/AKT) and endothelial nitric oxide synthase (eNOS) in each group were evaluated via Western blotting. The differences in the intracellular nitric oxide of RCECs treated by MF or ICA â ¡ were detected by DAF-FM DA that served as a nitric oxide fluorescent probe. Effects of MF and ICA â ¡ on cell proliferation of PA-stimulated RCECs were determined via CCK-8 assay. RESULTS: The content of nitric oxide in RCECs was significantly increased after the treatment of MF and ICA â ¡ in comparison with the NC group (P < 0.05). Moreover, compared with ICA â ¡ group, MF demonstrated a more obvious effect in promoting nitric oxide production (P < 0.05). Compared with the NC group, the expression levels of eNOS and AKT in the PA group were significantly decreased, indicating that a model for simulating the high-fat environment in vitro was successfully constructed (P < 0.05). Meanwhile, the intervention of MF and ICA â ¡ could effectively increase the expression of eNOS and AKT, suggesting that MF and ICA â ¡ could promote the recovery of endothelial dysfunction caused by high levels of free fatty acids (P < 0.05). The results of CCK-8 assays showed that PA could significantly reduce the proli-feration ability of RCECs (P < 0.05). Furthermore, the decreased cell viability induced by PA was significantly elevated by treatment with ICA â ¡ and MF (P < 0.05). CONCLUSION: In RCECs, MF and ICA â ¡ could effectively increase the content of nitric oxide. The down-regulation of the expression of proteins associated with the AKT/eNOS pathway after PA treatment revealed that this pathway was involved in the development of endothelial dysfunction, which could be effectively reversed by MF and ICA â ¡. In addition, the cell proliferation ability was significantly decreased following PA treatment, but MF and ICA â ¡ could restore the above changes. Overall, biflavonoid MF has an obvious repairing effect on PA-stimulated endothelial dysfunction.
Asunto(s)
Biflavonoides , Proteínas Proto-Oncogénicas c-akt , Animales , Biflavonoides/farmacología , Células Cultivadas , Células Endoteliales/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico/farmacología , Óxido Nítrico Sintasa de Tipo III/metabolismo , Óxido Nítrico Sintasa de Tipo III/farmacología , Ácido Palmítico/farmacología , Fosforilación , Proteínas Proto-Oncogénicas c-akt/metabolismo , Ratas , Transducción de SeñalRESUMEN
OBJECTIVE: To study the feasibility of transplantation of normal rat penile corpus cavernosum and major pelvic ganglion (MPG) into the renal subserous region of a Nu/Nu mouse based on allograft technology. METHODS: Penile corpus cavernosum and MPG, harvested from Sprague-Dawley (SD) rats under sterile condition, were transplanted underneath the kidney capsule of Nu/Nu mice through the microsurgery instruments and surgery microscope. The histopathologic changes and cellular proliferation in the transplanted penile corpus cavernosum and MPG were then analyzed at the end of 1week and 4 weeks after transplantation. Histological staining and immunohistochemical staining were used to evaluate the main outcome measures. RESULTS: After 1 week, the tissue morphology of the transplanted corpus cavernosum underneath the kidney capsule of Nu/Nu mice was consistent with normal penile corpus cavernosum, and blood could be observed in the penis cavernous sinus of the graft; after 4 weeks, the mophorlogy of the tranplanted corpus cavernosum near the kidney was consistent with normal penile corpus cavernosum, while fibrosis was noteworthy in the graft away from the kidney, but blood could still be seen in the penis cavernous sinus. After 1 week, the tissue morphology of the transplanted MPG was consistent with normal MPG, multiple islet-like cell clusters could be seen in the transplanted MPG in the renal subserous region, and angiogenesis could be observed near the kidney; after 4 weeks, a network of blood vessels was clearly visible away from the kidney, and islet-like cell clusters were still clearly observed in the transplanted MPG. In addition, ki67 positive cells were observed in the transplanted penile corpus cavernosum and MPG after 4 weeks of transplantation, which indicated that there was still cell proliferation activity in the grafts. CONCLUSION: The transplanted corpus cavernosum and MPG underneath the kidney capsule of Nu/Nu mice could survive at least 4 weeks. Moreover, the inner structure of the transplanted corpus cavernosum and MPG was close to the normal tissue. The underlining mechanism may be related to the local microenvironment underneath the kidney capsule of Nu/Nu mice and the neovascularization in the transplanted grafts.
RESUMEN
OBJECTIVE: To study the feasibility of transplantation of normal rat penile corpus cavernosum and major pelvic ganglion (MPG) into the renal subserous region of a Nu/Nu mouse based on allograft technology. METHODS: Penile corpus cavernosum and MPG, harvested from Sprague-Dawley (SD) rats under sterile condition, were transplanted underneath the kidney capsule of Nu/Nu mice through the microsurgery instruments and surgery microscope. The histopathologic changes and cellular proliferation in the transplanted penile corpus cavernosum and MPG were then analyzed at the end of 1week and 4 weeks after transplantation. Histological staining and immunohistochemical staining were used to evaluate the main outcome measures. RESULTS: After 1 week, the tissue morphology of the transplanted corpus cavernosum underneath the kidney capsule of Nu/Nu mice was consistent with normal penile corpus cavernosum, and blood could be observed in the penis cavernous sinus of the graft; after 4 weeks, the mophorlogy of the tranplanted corpus cavernosum near the kidney was consistent with normal penile corpus cavernosum, while fibrosis was noteworthy in the graft away from the kidney, but blood could still be seen in the penis cavernous sinus. After 1 week, the tissue morphology of the transplanted MPG was consistent with normal MPG, multiple islet-like cell clusters could be seen in the transplanted MPG in the renal subserous region, and angiogenesis could be observed near the kidney; after 4 weeks, a network of blood vessels was clearly visible away from the kidney, and islet-like cell clusters were still clearly observed in the transplanted MPG. In addition, ki67 positive cells were observed in the transplanted penile corpus cavernosum and MPG after 4 weeks of transplantation, which indicated that there was still cell proliferation activity in the grafts. CONCLUSION: The transplanted corpus cavernosum and MPG underneath the kidney capsule of Nu/Nu mice could survive at least 4 weeks. Moreover, the inner structure of the transplanted corpus cavernosum and MPG was close to the normal tissue. The underlining mechanism may be related to the local microenvironment underneath the kidney capsule of Nu/Nu mice and the neovascularization in the transplanted grafts.