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OBJECT IVE To study the effects of ergosterol peroxide derivatives EP-3P on the proliferation ，migration and invasion of human tripe negative breast cancer cell MDA-MB- 231，and to provide reference for the development of breast cancer related drugs. METHODS MTT assay was adopted to detect the proliferation of MDA-MB- 231 cells after treated with 0（blank control），1.25，2.5，5，10，20，40 μmol/L EP-3P for 24，48 and 72 h. Wound healing assay and Transwell chamber method were adopted to detect the migration and invasion ability of MDA-MB- 231 cells after treated with 0（blank control ），5，10，20 EP-3P for 24 h. The apoptosis and cell cycle distribution were detected by flow cytometry. Western blot assay was used to detect the expressions of B-cell lympho ma-2（Bcl-2），Bcl-2 associated X protein （Bax），caspase-3，cleaved-caspase-3，cytochrome C （Cyt-C），matrix metalloproteinase- 2（MMP-2）and MMP- 9. RESULTS Compared with blank control group ，2.5，5，10，20，40 μmol/L EP-3P could significantly increase the inhibitory rate of cell proliferation （P＜0.05 or P＜0.01）in a dose and time- dependent manner. After 24 h treatment of EP- 3P（10，20 μmol/L），the rate of cell migration and the number of invasive cells were decreased significantly （P＜0.01），and cell was arrested at G 2/M stage （P＜0.05 or P＜0.01）；the apoptotic rate was increased significantly （P＜0.05）；the protein expressions of Bax ，Cyt-C and cleaved-caspase- 3 were upregulated significantly ， while those of Bcl- 2，caspase-3，MMP-2 and MMP- 9 were downregulated significantly （P＜0.01）. CONCLUSIONS EP-3P can inhibit the proliferation ，migration and invasion of human tripe negative breast cancer cells MDA-MB- 231 through mitochondrial mediated endogenous caspase pathway ，and induce the apoptosis of cells .
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Objective To establish a mouse model of H.pylori infection, and to evaluate the chronic pathological changes in the gastric mucosa associated with H.pylori infection.Methods 34 male 5~6-week old SPF C57BL/6 mice were used in this study.The mice were intragastrically administrated with a suspension of H.pylori SS1 strain.Two weeks after infection, rapid urease test and PCR were performed to confirm the H.pylori infection.Successfully infected mice were randomly divided into 3 groups including the control group, 6-week and 12-week infected groups.Samples of gastric mucosa were taken for pathological analysis using HE and borax methylene blue staining.The contents of myeloperoxidase (MPO), superoxide dismutase (SOD), malondialdehyde (MDA) and catalase (CAT) in the gastric tissues were detected by biochemistry, and the expression levels of COX-2, iNOS, TNF-α and IL-1β were examined by RT-qPCR.Results Compared with the control group, H.pylori colonization was observed in the gastric mucosa of the 6-week and 12-week infected mice, with chronic inflammatory cell infiltration, glandular atrophy and intestinal metaplasia to varying extents.The contents of CAT and SOD were significantly decreased, while the levels of MPO and malonaldehyde MDA, and the expression levels of COX-2,iNOS,TNF-α and IL-1β were significantly increased (P<0.05 or P<0.01).Conclusions Intragastric administration with H.pylori in C57BL/6 mice can be successfully used to generate the bacterial colonization, leading to chronic inflammatory cell infiltration, enhanced oxidative stress, and up-regulated expression of proinflammatory genes in the gastric glandular tissues at 6 and 12 weeks after inoculation.However, the inflammatory changes are more extensive in the mice at 12 weeks after infection, with glandular atrophy and intestinal metaplasia.
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AIM To speculate the hypoglycemic mechanism for rats with type 2 diabetes by exploring the therapeutic effects of leaf aqueous extract from Cyclocarya paliurus on liver insulin receptor (InsR) and insulin receptor substrate 2 (IRS-2).METHODS The diabetic rat model was established through intraperitoneal injection of streptozotocin and fed with high-fat diet.The moleled rats were equally assigned into the control group and leaf aqueous extract from Cyclocarya paliurus group (extract group).After the test extract was orally administrated for four weeks,body weight,urine output,food intake,water intake and fasting blood-glucose (FBG) were measured,and the levels of serum insulin,InsR and IRS-2 mRNA in liver tissue were investigated in rats.RESULTS Compared with the control group,the extract group showed a reduction in urine output,food intake,water intake,FBG and insulin levels.Meanwhile,the rats' body weights in extract group were presented a trend to increase.The gene expressions of InsR and IRS-2 in liver tissue were up-regulated.Moreover,the insulin sensitivity was improved.CONCLUSION The leaf aqueous extract from Cyclocarya paliurus can reduce FBS,improve insulin sensitivity,which may be associated with the increase of InsR and IRS-2 gene expression in liver tissue.