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OBJECTIVE@#To investigate the effects of Korean Magnolia obovata crude extract (KME) on plateletderived growth factor (PDGF)-BB-induced proliferation and migration of vascular smooth muscle cells (VSMCs).@*METHODS@#KME composition was analyzed by high-performance liquid chromatography (HPLC). VSMCs were isolated from the aorta of a Sprague-Dawley rat, incubated in serum free-Dulbecco's modified Eagle's medium in the presence or absence of KME (10, 30, 100, and 300 μg/mL), then further treated with PDGF-BB (10 ng/mL). VSMC proliferation was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and VSMC migration was determined using the Boyden chamber and scratch wound healing assays. Western blot analysis was used to detect phosphorylation of extracellular signal-regulated protein kinases 1 and 2 (p-ERK1/2), protein kinase B (p-Akt), and stress-activated protein kinase/c-Jun NH2-terminal kinase (p-SAPK/JNK). The antimigration and proliferation effects of KME were tested using aortic sprout outgrowth.@*RESULTS@#The HPLC analysis identified honokiol (0.45 mg/g) and magnolol (0.34 mg/g) as the major components of KME. KME (30, 100, and 300 μg/mL) significantly decreased the proliferation and migration of PDGF-BB-stimulated (10 ng/mL) VSMCs and the PDGF-BB-induced phosphorylation of EKR1/2, Akt, and SAPK/JNK (P<0.05). Furthermore, PDGF-BBinduced VSMCs treated with 300 μg/mL of KME showed reduction in aortic sprout outgrowth.@*CONCLUSION@#KME could inhibit abnormal proliferation and migration of VSMCs by down-regulating the phosphorylation of EKR1/2 and Akt. Thus, KME might be a functional food for preventing vascular disorders.
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Objective To confirm the influence on the path of signal transduction of ERK by BaiHui-QuBin Scalp-acupuncture in rats of experimental acute intercerebral hemorrhage.Methods 192 healthy Wistar male rats were selected by examination.The rats were assigned into 4 groups randomly:matched blank group,model group,pin-prick group,excitomotor group.The latter three groups contained 60 rats in each;another 12 rats were assigned as the matched blank group.All rats were made to be the intercerebral hemorrage model.The pinprick group was acupunc-tured on the BaihuipenetratingQubin.We observed the change surrounding the hamatoma of the proteinum expression of p-ERK of each group in deferent time phase point with the method of Western blot.Results All of the rats after being made model appeared the phenomenon of the expression of p-ERK of rat's brain tissue surrounding the hematoma and increased to the peak in 7 days.1 d, 2 d, 3 d, 7 d, compare with the model group [(0.10 ±0.04),(0.17 ±0.02),(0.09 ±0.05),(0.10 ±0.06)],the expressions of p-ERK of the pinprick group [(0.19 ±0.04),(0.23 ±0.07),(0.24 ±0.05),(0.25 ±0.06),q=11.45,P0.05).Conclusion The therapy of BaiHui-QuBin Scalp-acupuncture can activate the path of signal transduction of ERK,increase the expression of the proteinum p-ERK and so on,which have the function of protecting the nerve cell.
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Objective To explore the role of integrin αvβ6 in mediating the tolerance of gastric cancer AGS cells to 5-fluorouracil, and to determine whether direct β6-extracellular signal-regulated protein kinase(ERK) binding is involved. Methods Gastric cancer AGS cells in the logarithmic phase were incubated with either 5-fluorouracil for 24 hours ( control group), with 0.1 g/L of mouse anti-αvβ6 monoclonal antibody 10D5 for 6 hours and then with 5-fluorouracil for24 hours ( 10D5 group), with IgG2a and 5-fluorouracil ( IgG2a group), or with 5-fluorouracil and 20 μnol/L of ERK inhibitor PD98059 for 24 hours ( PD98059 group). Cell proliferation,apoptosis and the expression of Bcl-2 and caspase-3 protein were detected by MTT assay, flow cytometry and western blotting, respectively. All data were analysed by one-way analysis of variance and LSD test. Results The cell inhibition rates of the control group, 10D5 group, IgG2a group and PD98059 group were 28.1% ±2.7%,84.5% ± 1.6%, 31.4% ±5.2%, 86.7% ±5.2%, respectively, with a significant difference ( F = 342. 5, P ＜0.05). The apoptosis rates of the control group, 10D5 group, IgG2a group, and PD98059 group were 6.6% ±1.4%, 30.6% ± 2.4%, 8.1% ± 1.3%, 36.0% ±4.0%, respectively, with a significant difference among the four groups (F = 105.4, P ＜0.05 ). There was a significant difference in the expression of caspase-3 and Bcl-2 among the four groups (F=292.1, 181.6, P＜0.05). Conclusion Integrin αvβ6 can mediate the tolerance of gastric cancer AGS cells to 5-fluorouracil through direct β6-ERK binding.
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Objective To explore the change in gene expression of extracellular signal regulated protein kinase1 (erk1),erk2,p38MAPK and 3 c-Jun N- terminal kinases (jnk1,jnk2,jnk3) in fetal skin at different developmental stages and children skin. Methods After morphological characteristics of fetal skin at different developmental stages were examined histologically,gene expressions of erk1,erk2,p38MAPK,jnk1,jnk2 and jnk3 were determined with reverse transcription-polymerase chain reaction analysis (RT-PCR). Results Compared with early gestational fetal skin,the levels of gene expression of erk1 and erk2 showed no substantial change in late gestational fetal skin,while the contents of transcripts of p38MAPK and jnk1 were significantly decreased,the expressions of mRNA of jnk2 and jnk3 were obviously elevated. In children skin,gene expressions of erk2,p38MAPK and jnk1 were even more remarkably lowered. In contrary,gene expressions of jnk2 and jnk3 were marked enhanced. Conclusion The relative elevation of gene transcription of erk2 and p38MAPK and the inhibition of gene expression of jnk2 and jnk3 in fetal skin of earlier developmental stage might be related to fetal scarless healing.