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Objective @#To investigate the effect of ginsenoside Rg1 (GRg1) on human retinal microvascular endothelial cells (HRMECs) glycolysis by regulating pyruvate kinase M2 ( PKM2) expression.@*Methods @#HRMECs were cultured in vitro and divided into normal control (NC) group, high glucose (HG) group, high glucose + ginsenoside Rg1 (HG + GRg1) group, high glucose + ginsenoside Rg1 + low expression PKM2 ( HG + GRg1 + si-PKM2) group, and high glucose + ginsenoside Rg1 + overexpression PKM2 (HG + GRg1 + OE⁃PKM2) group. si-PKM2 and OE⁃PKM2 were transfected into HRMECs cells by cell transfection. The expression of PKM2 mRNA in HRMECs was detected by qRT⁃PCR. The expression levels of related proteins in HRMECs were detected by Western blot. The number of lumen formation in vitro was observed under an inverted microscope to quantify the angiogenesis ability. Cell culture medium of each group was collected, and glucose intake, lactate production and adenosine triphosphate(ATP)content were detected by glucose detection kit, lactate detection kit and ATP detection kit,re spectively. @*Results @#HG induced HRMECs significantly increased the number of blood vessel formation, glycolysis and PKM2 expression, while GRg1 treatment significantly reduced the number of blood vessel formation, glycolysis and PKM2 expression; transfection of si⁃PKM2 assisted the inhibitory effect of GRg1 on glycolysis and angiogenesis while transfection of OE⁃PKM2 interfered with the function of GRg1 . @*Conclusion @#GRg1 inhibits angiogenesis by inhibiting PKM2 to reduce glycolysis of HRMECs.
RESUMEN
Objective To preliminarily explore the feasibility of tree shrew as a new kind of animal model in research of amblyopia,to discuss the primary visual cortex plasticity mechanism of form deprivation in tree shrew,and to provide a theoretical basis for further understanding the mechanism of amblyopia formation and recovery.Methods Sixty 30-days old tree shrews were divided into five groups,12 in each group:the group A had the right eye sutured for 1 month;the group B had the right eye sutured for 2 months;the group C had the left eye sutured for 1 month and then opened and the righ eye was sutured for 1 month,in other words,the group C was performed by alternating suture;the tree shrews of control group 1(D1) were in the same age as the the group A,but fed in normal breedingenvironment;the tree shrews of control group 2(D2) were at the same age of groups B and C,but fed with a normal diet.Samples of the visual cortex were taken after the completion of modeling,and were processed to observe the histology and ultrastructure of the visual cortex,the neuron apoptosis,and the c-fos protein expression in the tree shrews of different groups.Results Damages to different degrees were found by histological and electron microscopic examination of the visual cortex in each experimental group,and they were more obvious in the group sutured for 2 months.TUNEL staining showed that there were no significant differences between the apoptosis in the experimental and control groups.The expression of c-fos mRNA and protein in the experimental groups was decreased,and it was the lowest in the group sutured for 2 months.There was a small increase in the c-fos expression after the alternate suture,and no significant difference of c-fos expression was found in the control groups.Conclusions Different degrees of deprivation amblyopia lead to different histopathological changes.There is a plasticity in the neurons affected by amblyopia.Tree shrew can be used as an ideal animal model for the studies of form deprivation amblyopia.