1.Investigation of mechanism of SchA to decreasing MPP+-induced SH-SY5Y cell damage
Pengyan JI ; Yan LI ; Wei ZHANG ; Shunli PENG ; Zhe HE
Chongqing Medicine 2014;(29):3932-3934
Objective To explore the possible mechanism of SchA ,which decreases MPP+induce SH-SY5Y cell damage .Meth-ods Cultured cells were divided into 5 groups ,one as control group ,cultured by free-blood serum media;the other 4 groups were treated with different concentrations of SchA(1 ,3 ,5 μmol/L) and MPP+ (1 mmol/L) for 48 h named model group ,1 ,3 ,5 μmol/L SchA group respetivly .The content of nitric oxide(NO) were measured by NO kit ;The expression levels of total Akt and p-Akt proteins were detected by Western blot .Results Compared with the control group ,the content of NO in group significantly in-creased after MPP+stimulating(P<0 .05);compared to the control group ,the content of NO in 5μmol/L SchA group significantly decreased(P<0 .05) .The expression levels of total Akt in all groups had no significant difference(P>0 .05) .The expression levels of p-Akt in model group significantly lowered ,while SchA(1、3、5 μmol/L) significantly increased the expression levels of p-Akt in comparision with cells in model group .Conclusion Decreasing MPP+ induced SH-SY5Y cell damage of SchA may be related to the content of NO and p-Akt expression .
2.Induction effect of NPPB on apoptosis of human glioma SHG-44 cells and its mechanism
Jing TIAN ; Ling QI ; Pengyan JI ; Nan SHEN ; Wanli CUI ; Chunyan WANG
Journal of Jilin University(Medicine Edition) 2016;42(4):637-641
Objective:To investigate the induction effect of NPPB,a chloride channel blocker,on the apoptosis of human glioma SHG-44 cells,and to explore its mechanism. Methods:The SHG-44 cells were cultured in vitro and divided into control group and NPPB groups (50,100,200 μmol· L-1 ).The cell viability was detected by MTT assay.The apoptotic rates were detected by flow cytometry.The expression levels of Bax, Bcl-2 and caspase-3 were detected by immunohistochemical analysis and Western blotting method.Results:Compared with control group,the cell viabilities of SHG-44 cells in 100 and 200 μmol·L-1 NPPB groups after treated for 24 and 48 h were decreased significantly (P < 0.01).The results of flow cytometry showed that the apoptotic rates of SHG-44 cells in 100 and 200 μmol·L-1 NPPB groups were 24.64% and 41.85%,and they were higher than that in control group (4.17%) (P <0. 01).The immunohistochemical analysis and Western blotting results showed that the expression levels of caspase-3 and Bax proteins in SHG-44 cells in 100 μmol · L-1 NPPB group were increased (P < 0.05 or P < 0. 01 ), and the expression level of Bcl-2 protein was decreased (P < 0.05 ). Conclusion:NPPB could induce the apoptosis of human glioma SHG-44 cells by the down-regulation of the expression of Bcl-2 and the up-regulation of the expression of Bax,and the activation of caspase-3.
3.Effect of component II of broccoli polypeptide on glioma cell apoptosis
Ling QI ; Junjie XU ; Donghai ZHAO ; Lei HAN ; Pengyan JI ; Weiyao WANG
Chinese Journal of Pathophysiology 2014;(9):1584-1589
AIM:To explore the effect of component II of broccoli polypeptide on the apoptosis in glioma cells . METHODS:Human glioma SHG-44 cells were cultured and divided into control group and 3, 10, 30 and 100 mg/L com-ponent II of broccoli polypeptide groups .Cell viability was detected by MTT assay .The apoptotic rates were examined by Annexin V/PI staining.The morphological changes of the cells were observed under inverted microscope .The protein ex-pression of Bax and Bcl-2 was detected by immunocytochemistry and Western blotting .The protein level of caspase-3 was also examined by Western blotting .RESULTS:Treatment with component II of broccoli polypeptide for 24 h, 48 h or 72 h induced significant inhibition of viability of SHG-44 cells in a time-and dose-dependent manner .The results of Annexin V/PI staining showed that the apoptotic rates were increased in treatment groups in a dose -dependent manner .The density of glioma cells was decreased after treated with increasing concentrations of the drug , and the apoptotic bodies were ob-served under inverted microscope at 72 h.The results of immunocytochemistry and Western blotting showed that the expres-sion of Bax protein was increased but Bcl-2 protein expression was decreased , and the ratio of Bax/Bcl-2 was increased sig-nificantly compared with control group (P<0.05 or P<0.01).The level of caspase-3 protein was increased in 30 and 100 mg/L component II of broccoli polypeptide groups compared with control group (P<0.01).CONCLUSION:The compo-nent II of broccoli polypeptide increases the ratio of Bax /Bcl-2 and activates caspase-3 protein, thus inducing the apoptosis of glioma cells.
4.Study on mechanism of total flavonoids from hemerocallis fulva on oxidative stress and hepatocyte apoptosis in alcoholic liver injury
Bo XU ; Yan LI ; Pengyan JI ; Ling QI ; Qian LU ; Weinan WU ; Nan SHEN
Chongqing Medicine 2017;46(10):1304-1307
Objective To study the influence of total flavonoids of hemerocallis fulva(TFHF) on hepatocyte apoptosis and related protein expression in mice with alcoholic hepatic injury.Methods A total of 40 mice were randomly divided into four groups:blank control,model control andsmall and high dose TFHF groups,10 cases in each group.The mice were given the continuous gavage administration for 7 d.Then the model group was given once gavage by 50% ethanol 12.0 mL/kg after 1 h of the last administration.The blank control group was given the equal volume of distilled water.The activity levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum as well as the superoxide dismutase(SOD) activity and malondialdehyde (MDA) content in liver tissue hemogenate were detected.Hematoxylin and Eosin(HE) staining was performed for observing the pathological changes of the liver tissue.The flow cytometer was used to test the apoptosis ratio in hepatocyte suspension.The expressions of caspase-3,Bcl-2 and Bax protein were detected by Western blot.Results The various TFHF groups could decrease the activities of ALT and AST in serum (P<0.05),while could decrease the MDA content in liver tissue hemogenate (P<0.01) and increased the SOD activity;the liver tissue pathological examination showed that the high dose TFHF group could make the liver cell degeneration,alleviated the necrosis degree and relieved the pathological change of hepatic tissue;compared with the model group,the hepatocyte apoptosis rate in each TFHF group was decreased significantly;Western blotting results showed that the caspase-3 protein level in each TFHF group was decreased,expression of Bcl-2 protein was increased,whereas which of Bax protein was decreased and Bax/Bcl-2 ratio was reduced.Conclnsion TFHF has obvious protective effect on mice acute hepatic injury induced by ethanol,and can inhibit the hepatocyte apptosis,its action mechanism may be related to its antioxidation and regulation of caspase-3,Bcl-2 and Bax expression.
5.Effects of chronic intermittent hypobaric hypoxia on expression and promoter region methylation of key enzyme genes related to glucose metabolism in diabetic mice
Chunhong SUI ; Yantao HE ; Yawei XU ; Pengyan JI ; Ying CHANG ; Dongfang ZHANG ; Donghai ZHAO ; Lianhai JIN ; Cheng WANG
Journal of Environmental and Occupational Medicine 2024;41(8):911-918
Background Chronic intermittent hypobaric hypoxia (CIHH) can effectively alleviate type 2 diabetes mellitus (T2DM). In this process, the underlying mechanism in its association with the epigenetic regulation of DNA methylation in the promoter regions of glucose metabolism key enzyme genes remains unclear yet. Objective To investigate the effects of CIHH on expression and promoter region methylation of key enzyme genes related to glucose metabolism in diabetes mice, and to explore the underlying mechanism by which CIHH regulates glucose metabolism. Methods Forty C57BL/6J male mice were divided randomly into a normobaric normoxic control (NN/CON) group, a chronic intermittent hypobaric hypoxia intervention control (CIHH/CON) group, a normobaric normoxic diabetic model (NN/DM) group, and a chronic intermittent hypobaric hypoxia intervention diabetic model (CIHH/DM) group. The mice in the NN/DM and the CIHH/DM groups were fed for 7 weeks with high-fat and high-sugar diet. Subsequently, these mice were intraperitoneally injected consecutively with 50 mmol·L−1 streptozotocin (STZ) for 5 d at a dose of 40 mg·kg−1 (body weight) per day to create T2DM model mice. The mice in the CIHH/DM and the CIHH/CON groups were intervened by simulating hypobaric hypoxia at