1.Role of PTEN in diabetes mellitus-induced reduction of hypoxic postconditioning-induced protection of cardiomyocytes: the relationship with GSK-3β-mediated mitochondrial apoptotic pathway
Rui XUE ; Zhongyuan XIA ; Shaoqing LEI ; Bo ZHAO ; Qingtao MENG ; Wei LI ; Bin ZHOU
Chinese Journal of Anesthesiology 2017;37(1):108-111
Objective To evaluate the role of phosphatase and tensin homologue deleted on chromosome 10 (PTEN) in diabetes mellitus-induced reduction of hypoxic postconditioning (HPO)-induced protection of cardiomyocytes and the relationship with glycogen synthase kinase-3β (GSK-3β)-mediated mitochondrial apoptotic pathway.Methods H9c2 cells incubated in high-glucose (30 mmol/L) medium for 24 h were divided into 6 groups (n =5 each) using a random number table:normoxia group (group N),hypoxia-reoxygenation (H/R) group,group HPO,PTEN gene silencing normoxia group (group P-N),PTEN gene silencing H/R group (group P-H/R),and PTEN gene silencing HPO group (group P-HPO).H9c2 cells were exposed to 95% N2-5% CO2 for 4 h followed by 2 h reoxygenation with 90% O2-10% CO2.HPO was induced by 3 cycles of 5 min reoxygenation followed by 5 min hypoxia before reoxygenation.At the end of reoxygenation,the level of lactate dehydrogenase (LDH) in the supernatant was detected by enzyme-linked immunosorbent assay,the changes in mitochondrial membrane potential (MMP were assessed by JC-1 fluorescence assay,the cell apoptosis was detected by AnnexinV-FITC/PI flow cytometry,and the expression of PTEN and phosphorylated GSK-3β (p-GSK-3β) was determined by Western blot.The JC-1 monomer/polymer ratio and apoptosis rate were calculated.Results Compared with group N,the amount of LDH released,JC-1 monomer/polymer ratio and apoptosis rate were significantly increased,and the expression of PTEN was up-regulated in H/R and HPO groups (P<0.05).There was no significant difference in the parameters mentioned above between group H/R and group HPO (P>0.05).Compared with group HPO,the amount of LDH released,JC-1 monomer/polymer ratio and apoptosis rate were significantly decreased,PTEN expression was down-regulated,and the expression of p-GSK-3β was up-regulated in group P-HPO (P<0.05).Compared with group N,the expression of PTEN was significantly down-regulated,and no significant changes were found in the other parameters mentioned above in group P-N (P>0.05).Compared with group H/R,the expression of PTEN was significantly down-regulated,and no significant changes were found in the other parameters mentioned above in group P-H/R (P>0.05).Conclusion PTEN is involved in diabetes mellitus-induced reduction of HPO-induced protection of cardiomyocytes,and the mechanism is associated with PTEN-induced activation of GSK-3β-modulated mitochondrial apoptotic pathway.
2.Expression of PTEN in Myocardial Tissue in Coronary Heart Disease
Xuerong LI ; Yong HE ; Yujia LEI ; Xihe QIN ; Qingtao WEI ; Xinmin PAN ; Lijuan LI ; Lin ZHANG
Journal of Forensic Medicine 2016;32(2):94-96,104
Objective To observe the expression of phosphatase and tensin hom ology deleted on chrom o-som e ten (PTEN) in m yocardial tissue in patients w ith coronary heart disease, and explore the relevance betw een the expression of PTEN and the occurrence and developm ent of coronary heart disease. Methods A total of 16 death cases w ith pathological diagnosis of coronary heart disease w ere collected as experi-m ental group, and 19 cases w ithout m yocardial lesions w ere selected as control group. The expression of PTENprotein and its m RNA w ere detected by im m unohistochem istry and real-tim e fluorescence quanti-tative PC R respectively. The correlation betw een the expression of PTEN and the pathogenesis of coronary heart disease w as analyzed. Results The expression of PTENprotein in myocardium in cases w ith coro-nary heart disease w as significantly low er com pared w ith the control group (P<0.05). There w as no sta-tistical difference of the expression of PTEN m RNA betw een experim ental and control group (P>0.05). Conclusion PTEN m ay be involved in the occurrence and developm ent of coronary heart disease.
3.Treatment of post-stroke dysphagia by vitalstim therapy coupled with conventional swallowing training.
Wenguang XIA ; Chanjuan ZHENG ; Qingtao LEI ; Zhouping TANG ; Qiang, HUA ; Yangpu ZHANG ; Suiqiang ZHU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(1):73-6
To investigate the effects of VitalStim therapy coupled with conventional swallowing training on recovery of post-stroke dysphagia, a total of 120 patients with post-stroke dysphagia were randomly and evenly divided into three groups: conventional swallowing therapy group, VitalStim therapy group, and VitalStim therapy plus conventional swallowing therapy group. Prior to and after the treatment, signals of surface electromyography (sEMG) of swallowing muscles were detected, swallowing function was evaluated by using the Standardized Swallowing Assessment (SSA) and Videofluoroscopic Swallowing Study (VFSS) tests, and swallowing-related quality of life (SWAL-QOL) was evaluated using the SWAL-QOL questionnaire. There were significant differences in sEMG value, SSA, VFSS, and SWAL-QOL scores in each group between prior to and after treatment. After 4-week treatment, sEMG value, SSA, VFSS and SWAL-QOL scores were significantly greater in the VitalStim therapy plus conventional swallowing training group than in the conventional swallowing training group and VitalStim therapy group, but no significant difference existed between conventional swallowing therapy group and VitalStim therapy group. It was concluded that VitalStim therapy coupled with conventional swallowing training was conducive to recovery of post-stroke dysphagia.
4.Role of caveolin-1 in penehyclidine hydrochioride-induced inhibition of lipopolysaccharide-induced activation of TLR4/p38 MAPK signaling pathway in macrophages of mice
Qian KONG ; Xiaojing WU ; Lei ZHANG ; Bo ZHAO ; Ying JIANG ; Qingtao MENG ; Liying ZHAN ; Eyou WANG ; Zhongyua XIA
Chinese Journal of Anesthesiology 2017;37(9):1132-1135
Objective To evaluate the role of caveolin-1 (Cav-1) in penehyclidine hydrochioride (PHC)-induced inhibition of lipopolysaccharide(LPS)-induced activation of Toll-like receptor 4 (TLR4) /p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway in macrophages of mice.Methods Macrophages of mice were seeded in 6 em diameter dishes (5 ml per dish) and divided into 5 groups (n=20 each) using a random number table:Scr-siRNA group (S group),Scr-siRNA + LPS group (LPS group),Ser-siRNA+LPS +PHC group (LPS+P group),Cav-1-siRNA+LPS group (C+LPS group) and Cav-1-siRNA+LPS+PHC group (C+LPS+P group).Macrophages were transfected with Scr-siRNA for 24 h in S,LPS and LPS+P groups and with Smart pool Cav-1 siRNAs for 24 h in C+LPS and C+LPS+P groups.LPS at the final concentration of 1 μg/ml was added after the end of transfection,and macrophages were then incubated for 2 h in LPS,LPS+P,C+LPS and C+LPS+P groups.In LPS+P and C+LPS+P groups,PHC at the final concentration of 2 μg/ml was added at 2 h of incubation with LPS,and macrophages were then incubated for 2 h.The expression of Cav-1 and TLR4 was detected by Western blot.The expression of p38 MAPK was determined by immunofluorescence.The level of tumor necrosis factor-alpha (TNF-α) in the culture medium was determined by enzyme-linked immunosorbent assay.The activity of myeloperoxidase (MPO) in macrophages was measured by colorimetry.Results Compared with group S,the expression of TLR4 and p38 MAPK was significantly up-regulated,and the concentration of TNF-α in the culture medium and activity of MPO were increased in the other four groups,the expression of Cav-1 was significantly downregulated in LPS and C+LPS groups (P <0.05),and no significant change was found in the expression of Cav-1 in group LPS+P (P>0.05).Compared with group LPS,the expression of Cav-1 was significantly up-regulated,the expression of TLR4 and p38 MAPK was down-regulated,and the concentration of TNF-α in the culture medium and activity of MPO were decreased in group LPS+P,and the expression of Cay-1 was significantly down-regulated,the expression of TLR4 and p38 MAPK was up-regulated,and the concentration of TNF-α in the culture medium and activity of MPO were increased in group C+LPS (P<0.05).Compared with group LPS+P,the expression of Cav-1 was significantly down-regulated,the expression of TLR4 and p38 MAPK was up-regulated,and the concentration of TNF-α in the culture medium and activity of MPO were increased in group C+LPS+P (P<0.05).Compared with group C+LPS,the expression of Cav-1 was significantly up-regulated,the expression of TLR4 and p38 MAPK was down-regulated,and the concentration of TNF-α in the culture medium and activity of MPO were decreased in group C+LPS+P (P<0.05).Conclusion The mechanism by which PHC inhibits LPS-induced activation of TLR4/p38 MAPK signaling pathway in macrophages is related to up-regulating Cav-1 expression in mice.
5.Treatment of Post-stroke Dysphagia by VitalStim Therapy Coupled with Conventional Swallowing Training
XIA WENGUANG ; ZHENG CHANJUAN ; LEI QINGTAO ; TANG ZHOUPING ; HUA QIANG ; ZHANG YANGPU ; ZHU SUIQIANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(1):73-76
To investigate the effects of VitalStim therapy coupled with conventional swallowing training on recovery of post-stroke dysphagia,a total of 120 patients with post-stroke dysphagia were randomly and evenly divided into three groups:conventional swallowing therapy group,VitalStim therapy group,and VitalStim therapy plus conventional swallowing therapy group.Prior to and after the treatment,signals of surface electromyography (sEMG) of swallowing muscles were detected,swallowing function was evaluated by using the Standardized Swallowing Assessment (SSA) and Videofluoroscopic Swallowing Study (VFSS) tests,and swallowing-related quality of life (SWAL-QOL) was evaluated using the SWAL-QOL questionnaire.There were significant differences in sEMG value,SSA,VFSS,and SWAL-QOL scores in each group between prior to and after treatment.After 4-week treatment,sEMG value,SSA,VFSS and SWAL-QOL scores were significantly greater in the VitalStim therapy plus conventional swallowing training group than in the conventional swallowing training group and VitalStim therapy group,but no significant difference existed between conventional swallowing therapy group and VitalStim therapy group.It was concluded that VitalStim therapy coupled with conventional swallowing training was conducive to recovery of post-stroke dysphagia.