RESUMO
@#AIM:To investigate the effect of silencing glial fibrillary acidic protein gene(GFAP)on proliferation and apoptosis of high glucose-induced human retinal microvascular endothelial cells(hRMECs)and its mechanism.<p>METHODS: Expression of GFAP in hRMECs treated with high sugar(30mmol/mL)and low sugar(5mmol/mL)was detected by qRT-PCR. The high glucose-induced hRMECs cells of silencing GFAP gene was established by lentiviral-mediated method. High glucose-induced hRMECs cells were treated with SRI-011381(TGF-β signaling pathway activator)and dimethyl sulfoxide(DMSO); Expression of GFAP, transforming growth factor-1(TGF-β1), activating transcription factor2(Smad2), Smad3 proteins were measured by Western blot, and cell proliferation and apoptosis were detected by CCK-8 and flow cytometry, respectively.<p>RESULTS: Expression of GFAP was significantly increased in high glucose treated hRMECs. The high glucose induced hRMECs cell model of GFAP gene silencing was successfully constructed by lentivirus mediation, and the cell proliferation ability was significantly improved, the apoptosis rate is significantly inhibited, and expression of TGF-1, Smad2 and Smad3 proteins in the TGF-β signaling pathway was significantly inhibited after silencing GFAP, while activation of TGF-β signaling pathway could reverse the inhibitory effect of silencing GFAP on the proliferation and apoptosis in high glucose hRMECs.<p>CONCLUSION: Silencing GFAP gene can promote the proliferation of high glucoseinducedhuman retinal microvascular endothelial cells and inhibit cell apoptosis,the mechanism may be related to the inactivation of TGF-β signaling pathway.
RESUMO
Objective To analyze the clinical and MRI features of patients with type Ⅱ Alexander disease (AxD) in order to better understand and diagnose it earlier.Methods Four type Ⅱ AxD patients identified by glial fibrillary acidic protein gene mutations from Peking University First Hospital and 128 type Ⅱ AxD cases from published literatures were collected,and the clinical and MRI features were summarized.Results (1) In 4 type Ⅱ AxD patients,2 adult patients showed abnormal MRI features without clinical manifestation.The other 2 children patients both manifested motor dysfunction of lower limbs,pyramidal signs,paroxysmal deterioration,and seizures during the course of disease,while 1 of them had bulbar paralysis.The MRI of all the cases was abnormal,but only 1 case MRI corresponded with typical MRI features of type Ⅱ AxD.In the other 3 cases MRI showed thc atrophy in the medulla and upper spinal cord,or the brainstem lesions and abnormal signal in the periventricular white matter,and abnormal basal ganglia region.(2) In 128 reported type Ⅱ AxD cases,the age of onset was (32±19) years old.The initial syndromes mainly contained bulbar and/or pseudobulbar paralysis (32.48％,38/117 cases),motor dysfunction of the lower limbs (31.62％,37/117 cases) and autonomic nerve dysfunction (13.67％,16/117 cases).During the course of the disease,the clinical manifestation showed bulbar and/or pseudobulbar paralysis (73.50％,86/117 cases),pyramidal signs (60.68％,71/117 cases) and ataxia (51.28％,60/117 cases).The MRI of all cases was characterized by atrophy or abnormal signals in the brainstem,especially in medulla oblongata,and spinal cord.And abnormal signals in the cerebellar dentate nuclei,white matter,basal ganglia and thalamus were also commonly shown in the MRI.Conclusions The patients with type Ⅱ AxD are late-onsct.The clinical manifestation mainly contains bulbar and/or pseudobulbar paralysis,motor dysfunction of the lower limbs and pyramidal signs.The MRI is characterized by atrophy or abnormal signals in the brainstem (especially in medulla oblongata) and spinal cord.