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Objective To research the effect and mechanism of phosphatidylinositol 3‐kinase(PI3K)/serine threonine protein kinase(AKT) and mitogen extracellular signal‐regulated kinase(MEK)/extracellular signal‐regulated kinase(ERK) signaling path‐way in tumor vascular endothelial cell migration .Methods The different concentrations of PI3K/AKT signaling pathway inhibitor LY294002 (2 .50 ,7 .50 ,15 .00 μmol/L) and the MEK/ERK signaling pathway inhibitor PD98059 (2 .50 ,7 .50 ,15 .00 μmol/L) were used to treat the tumor‐derived endothelial cells respectively .The DMEM‐F12 culture medium was added into 0 .10% DMSO culture medium as the control .The cell scratch test ,cell directional migration test and Transwell test were adopted to detect the effects of different signaling pathway inhibitors on the tumor vascular endothelial cells level ,horizontal ,vertical and directional mi‐gration .Results 0 .10% DMSO had no significant effect on endothelial cell migration and the role of endothelial cells′migration a‐bility after its action had no obvious difference compared with the single DMEM‐F12 medium action ,indicating that small doses of DMSO as the solvent of LY294002 ,PD98059 did not affect the tumor vascular endothelial cell migration function ;after treatment by signaling pathway inhibitor LY294002 and PD98059 ,the endothelial cell migration ability was suppressed and increased with the in‐hibitor concentration increase ;compared with the PD98059 group ,the migration distance in the LY294002 group was smaller and the number of migrating cells was less ,the differences had statistical significance (P<0 .05) .Conclusion The inhibition of PI3K/AKT and MEK/ERK signaling pathway can inhibit the level of tumor vascular endothelial cells ,vertical and directional migration , and which is positively correlated with the concentration of inhibitor ;the effect of PI3K/AKT signal pathway on the migration of endothelial cells is more significant than that of the MEK/ERK signal pathway .
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Objective To investigate that ox-LDL inhibits endothelial differentiation of bone marrow mesenchymal stem cells (MSCs) in rats and its underlying mechanism .Methods Cultured MSCs were divided into four groups:blank groups , ox-LDL groups ( 5 μg/mL ox-LDL ) , ox-LDL +NAC groups ( 5 μg/mL ox-LDL and pre-treated NAC), and negative LDL groups (5 μg/mL nLDL).Cell morphology, endothelial marker and differentiation effi-ciency as well as signal of oxidative and pathway protein were detected after induction by Western blot , real-time PCR.Results MSCs can differentiate into endothelial cells with the expression of endothelial marker vWF , Flk-1 and CD31 at the mRNA and protein level , vascular morphology , ox-LDL obvious inhibited endothelial differentia-tion of MSCs ( P<0.05 ) , but NAC can reverse the inhibition , the amount of ROS in ox-LDL groups was higher than that in ox-LDL+NAC groups ( P <0.05 ) , The expression of phosphorylated Akt decreased distinctly after treatment with ox-LDL( P<0.05 ) , NAC can stimulated its expression close to normal .Conclusions ox-LDL can inhibit endothelial differentiation of MSCs via ROS , NAC in this procese shows inhibition to effect of ox-LDL and Akt signaling also played a critical role .
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Objective To research the effect mechanism of ultrashortwave in the correlation of ultrashortwave and the tail replantation, provide the experiment basis of clinical practice of prevention and cure for the vascular crisis after micromodule anastomosis. Methods Eighty Sprague-Dawley(SD) rats of clean grade were 3-month-old,female,and were divided into four groups:control group (group 0),model group (group 1 ),contrast group (group 2),ultrashortwave (USW) group (group 3).The preparation of tail replantation model was cut off soft tissue except for caudal veins on both sides of the tail. The coccyx was not broken away from tail.At last,the audal artery under abdominal main centre ditch was anastomosed.In experiment process, the USW group was divided into high dosage group (group 3A) and low dosage group (group 3B). The caudal arterys were ligated and not anastomosed in the group 0. Caudal arterys in other groups were anastomosed.Rats in the group 0 and group 1 received no treatment,normal management after the operation. Rats in the group 2 were given abdominal cavity injection of papaverin liquid immediately,then once a day to 5 days after the tail replantation.Rats in the group 3 were immediately given USW therapy of twenty minutes on the anastomosis section,and then once a day for 5 days after the tail replantation.The USW dosage of group 3A was 3th grade and 50mA. The USW dosage of group 3B was 2th grade and 28mA.The survival rate of the rat tails was observed after the tail replantation for 10 day.Before being grouped,it was measured that the tail skin temperature diference between near and far side of anastomosis section.After the tail relpantation, the temperature diference was inspected daily for 10 postoperative days hence. Before rats were grouped and the eighth hour after the tail replantation, rats were collected blood plasma specimens and measured contents of nitric oxide with destination colorimetric mathods of nitric oxide.Results Carrying out comparison of survival rate of every group,the output weve:between tail cutting off group (group 0,0) and tail replantation group (group 1,2,3,43.94％) to compare P ＞ 0.05; between each group of the tail replantation groups (1,2,3A,3B group) to compare P＞ 0.05,group 3B ＜ 2 ＜ 1 ＜ 3A; between group 3B and group 1 to compare P ＞ 0.05; between group 2 and group 1 to compare P ＞ 0.05. Each group were compared with the change daily between postoperative and preceding operative the skin temperature diference,single-factor analysis of variance (One-Way ANOVA) analysis:Postoperative 1 day,group 3A ＜ 1,P ＜ 0.05.Postoperative 6 day:3A ＜ 3B ＜ 1 ＜ 2,P ＞ 0.05.Postoperative 7 day:group 3B ＜ 1 ＜ 3A ＜ 2,P ＜ 0.05.Each group were compared with the change of the content of nitric oxide between postoperative 8 hour and preceding operative,with rank-sum test:group 3B ＞ 3A ＞ 2 ＞ 1 ＞ 0,H =33.760,P ＜ 0.05,shows statistically significant.Conclusions USW therapy,especially USW low-dose therapy,can reduce vascular crisis and improve the survival rate of replanted rat tails,after the postoperative 1,6,7 days,reduce skin temperature,improve blood supply,improve nitric oxide at postoperative eighth hour,prevent vascular crisis.Rat tail replantation model in this experiment is feasible.