1.Ocular findings and histological change in glaucoma model of DBA/2J Mice
Fan, YANG ; Lingling WU ; Xiujuan, GUO ; Liping, YANG ; Ying, LI ; Lemeng, WU ; Dongmei, WANG
Chinese Ophthalmic Research 2010;28(2):103-108
Background As a hereditary chronic glaucoma model,DBA/2J mouse is widely used in the experimental research of glaucoma worldwide.Although some researches have determined the ocular change induced by hypertention of DBA/2J mouse,there is seldom reports about the research on the relationship of ocular pathological abnormality and development course in DBA/2J mouse.Objective The present study is to characterize the ocular abnormalities and histological changes induced by hypertention in different ages of DBA/2J mice.Methods The clean female DBA/2J mice aged 3-,5-,7-,9-,11-,14-month-old (6 mice for each) were used in this study,and age matched 18 female C57BL/6J mice were as controls.Intraocular pressure (IOP) of mice was measured by anterior chamber puncture of microneedle.The animals were sacrificed and retinal flat mounts were prepared for histopathological examination under the light microscope.Retinal ganglion cells (RGCs) were counted by retinal Nissl staining.Morphology of optic nerve cup in frozen section was examined under the light microscope.The experiment followed the Standard of Association for Research in Vision and Ophthalmology.Results Developing pigment dispersion,iris stroma atrophy,transillumination defects and pupil deformation were found in DBA/2J mice.IOP began to rise in 7-month-old DBA/2J mouse,peaked in 9-month-old mouse and returned to normal in 14-month-old DBA/2J mouse.A significant difference was found in IOP among different ages of DBA/2J mice (F=27.600,P<0.05) but not C57BL/6J mice (F=0.249,P=0.781).RGCs loss was observed in 7-month-old DBA/2J mice and more serious from 9 to 11-month-old mice,showing a significant decline of RGCs among different ages of DBA/2J mice (F=23.594,P=0.000) but not C57BL/6J mice (F=1.816,P=0.211).The abnormality of optic nerve cupping and decrease of retinal nerve fiber layer were found in 9 to 14-month-old months old DBA/2J mice and were normal in age-matched C57BL/6J mice.Conclusion The abnormal alteration of the ocular anterior segment in DBA/2J mouse is gradually aggravated with aging.The findings of eye in DBA/2J mouse is characterized by secondary glaucoma.DBA/2J mouse offer an ideal animal model for the research of glaucoma.
2.Proliferation effects of berberine on rat neural stem cells induced by hydrogen peroxide
Shengnan SUN ; Xin LIU ; Miaomiao TONG ; Shuyan YI ; Lemeng FAN ; Shuang LIU ; Xinru LI ; Leqi FAN ; Nan ZHANG
Chinese Journal of Comparative Medicine 2018;28(5):60-64
Objective To investigate the effect of berberine on proliferation of neural stem cells(NSCs)induced by hydrogen peroxide(H2O2). Methods NSCs from Sprague-Dawley rats were isolated and purified by suspension culture. Cells were divided into a control group,H2O2group(NSCs exposed to H2O2injury),berberine group(NSCs were incubated with berberine concentrations ranging from 0.5 to 20 μmol/L and exposed to H2O2), and DAPT(a blocker of the Notch signaling pathway)group. Cell viability was evaluated using the Cell Counting Kit-8 assay. Proliferation of NSCs was evaluated by a neurosphere formation assay and Ki67 protein expression. Expression of key proteins in the Notch signaling pathway(including notch1 and hes1)in response to berberine treatment or DAPT(a Notch inhibitor)was determined by Western blotting. Results Cell viability of NSCs was significantly increased by berberine compared with the H2O2group. The neurosphere growth assay showed that 5 or 10 μmol/L berberine increased NSC proliferation. The ratio of Ki67 +/DAPI cells and notch1 and hes1 protein expression increased significantly compared with the H2O2group. Conclusions Berberine treatment upregulates Notch signaling in NSCs,whereas DAPT attenuates these effects. Berberine is a drug that promotes NSC proliferation and exerts a protective effect on NSCs via the Notch signaling pathway.