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Effects of high glucose and severe hypoxia on the biological behavior of mesenchymal stem cells at various passages

World Journal of Stem Cells 2024年第4期16卷 434-443页

作者： Fatimah Almahasneh Ejlal Abu-El-Rub Ramada R Khasawneh Rawan Almazari Department of Basic Medical Sciences Faculty of MedicineYarmouk UniversityIrbid 21163Jordan

BACKGROUND Mesenchymal stem cells(MSCs)have been extensively studied for therapeutic potential,due to their regenerative and immunomodulatory *** passage and stress factors may affect the biological characteristics of MSCs,but the details of these effects have not been recognized *** To investigate the effects of stress factors(high glucose and severe hypoxia)on the biological characteristics of MSCs at different passages,in order to optimize the therapeutic applications of *** In this study,we investigated the impact of two stress conditions;severe hypoxia and high glucose on human adipose-tissue derived MSCs(hAD-MSCs)at passages 6(P6),P8,and ***,senescence and apoptosis were evaluated measuring WST-1,senescence-associated beta-galactosidase,and annexin V,*** Cells at P6 showed decreased proliferation and increased apoptosis under conditions of high glucose and hypoxia compared to control,while the extent of senescence did not change significantly under stress *** P8 hAD-MSCs cultured in stress conditions had a significant decrease in proliferation and apoptosis and a significant increase in senescence compared to counterpart cells at *** cultured in high glucose at P10 had lower proliferation and higher senescence than their counterparts in the previous passage,while no change in apoptosis was *** the other hand,MSCs cultured under hypoxia showed decreased senescence,increased apoptosis and no significant change in proliferation when compared to the same conditions at *** These results indicate that stress factors had distinct effects on the biological processes of MSCs at different passages,and suggest that senescence may be a protective mechanism for MSCs to survive under stress conditions at higher passage numbers.

关键词： Mesenchymal stem cells high glucose Hypoxia Stressful microenvironment Serial passage Senescence Apoptosis

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high glucose levels impact visual response properties of retinal ganglion cells in C57 mice—An in vitro physiological study

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Science China(Life Sciences) 2017年第12期60卷 1428-1435页

作者： Yuan Zhou Chunxia Xiao Mingliang Pu Department of Anatomy/Embryology School of Basic Medical Sciences Peking University Functional Department Mawangdui Branch People's Hospital of Hunan Province The First Affiliated Hospital of Hunan Normal University

This study investigated visual response properties of retinal ganglion cells(RGCs) under high glucose levels. Extracellular single-unit responses of RGCs from mouse retinas were recorded. And the eyecup was prepared as a flat mount in a recording chamber and superfused with Ames medium. The averaged RF size of the ON RGCs(34.1±2.9, n=14) was significantly smaller than the OFF RGCs under the HG(49.3±0.3, n=12)(Phigher than OFF cells with the HG treatment(P<0.0001). The present results suggest that HG reduced receptive field center size, suppressed luminance threshold, and attenuated contrast gain of RGCs. The impact of HG on ON and OFF RGCs may be mediated via different mechanisms.

关键词： retinal ganglion cells high glucose high mannitol receptive field center size luminance threshold contrast threshold

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high glucose causes apoptosis of rabbit corneal epithelial cells involving activation of PERK-eIF2α-CHOP-caspase-12 signaling pathway

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International Journal of Ophthalmology(English edition) 2019年第12期12卷 1815-1822页

作者： Pan-Pan Yao Min-Jie Sheng Wen-Hao Weng Yin Long Hao Liu Li Chen Jia-Jun Lu Ao Rong Bing Li Department of Ophthalmology Yangpu HospitalTongji University School of MedicineShanghai 200090China Department of Clinical Laboratory Yangpu HospitalTongji University School of MedicineShanghai 200090China Department of Ophthalmology Tongji HospitalTongji University School of MedicineShanghai 200065China

AIM: To investigate the effect of high concentration of glucose(HCG) on double stranded RNA-activated protein kinase-like ER kinase(PERK)-eukaryotic initiation factor-2α(eIF2α)-transcription factor C/EBP homologous protein(CHOP)-cysteine aspartate specific proteinase(caspase-12) signaling pathway activation and apoptosis in rabbit corneal epithelial cells(RCECs). METHODS: RCECs were treated by different concentrations of glucose for 0-48 h. The expressions of PERK, p-PERK, eIF2α, p-eIF2α, 78 k Da glucose-regulated protein 78(GRP78), CHOP, B-cell lymphoma 2(Bcl-2), B-cell lymphoma-2-associated X protein(Bax) and caspase-12 were determined by Western blot. Apoptosis was detected by TUNEL assay. Meanwhile, the function of PERK-eI F2α-CHOP-caspase-12 signaling pathway activation in high glucose-induced apoptosis was evaluated using PERK inhibitor, GSK2606414. RESULTS: HCG significantly promoted the expression of p-PERK, p-eIF2α, GRP78, CHOP, Bax and cleaved caspase-12 in RCECs(Pglucose were in concentration-and time-dependent manners. Meanwhile, PERK and eIF2α expressions were not affected in all groups(P>0.05). TUNEL assay showed that the apoptosis rate of RCECs in the HCG group increased significantly in contrast with that in the normal concentration of glucose or osmotic pressure control group(Pglucose concentration within limits(P0.05). Correspondingly, GSK2606414 also significantly reduced the apoptosis rate induced by high glucose(P<0.05). CONCLUSION: HCG activates PERK-eIF2α-CHOPcaspase-12 signaling pathway and promotes apoptosis of RCECs.

关键词： high glucose rabbit corneal epithelial cells PERK-eIF2α-CHOP-caspase-12 pathway apoptosis

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high glucose Promotes the CTGF Expression in Human Mesangial Cells via Serum and Glucocorticoid-induced Kinase 1 Pathway

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2008年第5期28卷 508-512页

作者：王全胜张阿丽李仁康刘建国谢纪文邓安国冯玉锡朱忠华 Department of Integrated Chinese Medicine and Western Medicine Union Hospital TongJi Medical College Huazhong University of Science and Technology Department of Radiotherapy and Chemotherapy Zhongnan Hospital Wuhan University 3Department of Nephrology Union Hospital TongJi Medical College Huazhong University of Science and Technology

The role of serum and glucocorticoid-induced kinase 1 （SGK1） pathway in the connective tissue growth factor （CTGF） expression was investigated in cultured human mesangial cells （HMCs） under high glucose. By using RT-PCR and Western blot, the effect of SGK1 on the CTGF expression in HMCs under high glucose was examined. Overexpression of active SGK1 in HMCs transfected with PIRES2-EGFP- S422D hSGK1 （SD） could increase the expression of phosphorylated SGK1 and CTGF as compared with HMCs groups transfected with PIRES2-EGFP （FP） under high glucose or normal glucose. Overexpression of inactive SGK1 in HMCs transfected with PIRES2-EGFP- K127N hSGK1 （KN） could decrease phosphorylated SGK1 and CTGF expression as compared with HMCs groups transfected with FP under high glucose. In conclusion, these results suggest that high glucose-induced CTGF expression is mediated through the active SGK1 in HMCs.

关键词： high glucose serum and glucocorticoid-induced protein kinase 1 human mesangial cells connective tissue growth factor diabetic nephropathy

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high glucose induces myocardial cell injury through increasing reactive oxygen species production

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Asian Pacific Journal of Tropical Medicine 2018年第1期11卷 63-67页

作者： Yu-Jun Wang Xiao-Yu Lyu Li Yu Department of Critical Care Medicine the Central Hospital of WuhanTongji Medical CollegeHuazhong University of Science and Technology Department of Endocrinology the Central Hospital of WuhanTongji Medical CollegeHuazhong University of Science and Technology

Objective: To study the injury effect and molecular mechanism of high glucose on myocardial cells. Methods: Myocardial cells H9 c2 were cultured and divided into the control group treated with DMEM containing 5.5 mmol/L glucose, the high glucose group treated with DMEM containing 35 mmol/L glucose, and the N-acetylcysteine(NAC) group pre-treated with 1000μmol/L NAC and treated with DMEM containing 1000 μmol/L NAC and 35 mmol/L *** production of ROS and the expression of mitochondria pathway apoptosis molecules in cells as well as the contents of collagen and collagen metabolism molecules were ***: After 8 h, 16 h and 24 h of treatment, ROS RFU as well as Bax, CytC, Caspase-3 and Caspase-9 protein expression in cells and Col-I, Col-Ⅲ, PINP and PⅢNP protein levels in culture medium of high glucose group were higher than those of control group, Bcl-2 protein expression were lower than those of control group, but CTX-Ⅰ protein levels in culture medium were not significantly different from those of control group; after 24 h of treatment, Bax, CytC,Caspase-3 and Caspase-9 protein expression in cells as well as Col-Ⅰ, Col-Ⅲ, PINP and PIIINP protein levels in culture medium of NAC group were lower than those of high glucose group whereas Bcl-2 protein expression was higher than that of high glucose group. Conclusions:high glucose can induce myocardial cell apoptosis, increase collagen synthesis and accelerate interstitial fibrosis by increasing the production of reactive oxygen species.

关键词： Diabetic cardiomyopathy high glucose Myocardial cells Reactive oxygen species Apoptosis Collagen

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Anti-Oxidative and Anti-Inflammatory Effects of Cinnamaldehyde on Protecting high glucose-Induced Damage in Cultured Dorsal Root Ganglion Neurons of Rats

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Chinese Journal of Integrative Medicine 2016年第1期22卷 19-27页

作者：杨丹梁晓春石玥孙青刘頔刘伟张宏 Department of Traditional Chinese Medicine Peking Union Medical College HospitalPeking Union Medical CollegeTranslational Medicine CenterChinese Academy of Medical Sciences Department of Cell Resource Center Institute of Basic Medical SciencePeking Union Medical CollegeChinese Academy of Medical Sciences

Objective： To examine the mechanism underlying the beneficial role of cinnamaldehyde on oxidative damage and apoptosis in high glucose （HG）-induced dorsal root ganglion （DRG） neurons in vitro. Methods： HG-treated DRG neurons were developed as an in vitro model of diabetic neuropathy. The neurons were randomly divided into five groups： the control group, the HG group and the HG groups treated with 25, 50 and 100 nmol/L cinnamaldehyde, respectively. Cell viability was examined by 3-（4,5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide （MTT） assay and apoptosis rate was evaluated by the in situ TdT-mediated dUTP nick end labeling （TUNEL） assay. The intracellular level of reactive oxygen species （ROS） was measured with flow cytometry. Expression of nuclear factor-kappa B （NF-κ B）, inhibitor of κ B （I κ B）, phosphorylated I κ B （p-IκB）, tumor necrosis factor （TNF）-α, interleukin-6 （IL-6） and caspase-3 were determined by western blotting and real-time quantitative reverse transcription polymerase chain reaction （RT-PCR）. Expression of nuclear factor erythroid 2-related factor 2 （Nrf2） and hemeoxygenase-1 （HO-1） were also measured by western blotting. Results： Cinnamaldehyde reduced HG-induced loss of viability, apoptosis and intracellular generation of ROS in the DRG neurons via inhibiting NF- K B activity. The western blot assay results showed that the HG-induced elevated expressions of NF- κB, I κ B and p-I κ B were remarkably reduced by cinnamaldehyde treatment in a dose-dependent manner （P〈0.01）. The HG-induced over-expression of NF-κ B p65 mRNA was remarkably attenuated after cinnamaldehyde treatment in a dose-dependent manner （P〈0.01）. However, the expressions of Nrf2 and HO-1 were not upregulated. Treatment with cinnamaldehyde not only attenuated caspase-3 activation and the caspase cleavage cascade in DRG neurons, but also lowered the elevated IL-6, TNF-α, cyclo-oxygenase and inducible nitric oxide synthase levels, indicating a reduction in inflammatory dama

关键词： nuclear factor-kappa B nuclear factor erythroid 2-related factor 2 cinnamaldehyde high glucose dorsal root ganglion

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Benfotiamine prevents increased β-amyloid production in HEK cells induced by high glucose

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Neuroscience Bulletin 2012年第5期28卷 561-566页

作者： Xiao-Jing Sun Lei Zhao Na Zhao Xiao-Li Pan Guo-Qiang Fei Li-Rong Jin Chun-Jiu Zhong Department of Neurology Zhongshan Hospital & Shanghai Medical College Fudan University Shanghai 200032 China State Key Laboratory of Medical Neurobiology Institute of Brain Science Fudan University Shanghai 200032 China

Objective To determine whether high glucose enhances β-amyloid （Aβ） production in HEK293 Swedish mutant （APPsw） cells with Aβ precursor protein （APP） overexpression, and whether under this condition benfotiamine reduces the increased Aβ production. Methods HEK293 APPsw cells were cultured with different concentrations of glucose for different times. TheAβ content in the supernatant was determined by ELISA. To investigate the mechanism by which benfotiamine reduced Aβ production, glycogen synthase kinase-3 （GSK-3） activity and expression were measured after the cells were cultured with 5.5 g/L glucose for 12 h. Results With 1.0, 3.0, 4.5, 5.5, 6.5, 7.5, 8.5, or 10.5 g/L glucose, Aβ production by HEK293 APPsw cells was highest in the presence of 5.5 g/L glucose for 6 and 12 h. The difference in Aβ content between 5.5 and 1.0 g/L was most marked after incubation for 12 h. Benfotiamine at 20 and 40 μg/mL significantly reduced Aβ production in cells incubated with 5.5 g/L glucose for 12 h. Moreover, 40 μg/mL benfotiamine significantly enhanced the ratio of phosphorylated GSK-3 to total GSK-3, together with consistent down-regulation of GSK-3 activity. Conclusion high glucose increases Aβ production by HEK293 APPsw cells while benfotiamine prevents this increase. This is correlated with the modulation of GSK-3 activity.

关键词： Alzheimer’s disease β-amyloid high glucose benfotiamine glycogen synthase kinase

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Exosomes Derived from Human Umbilical Cord Mesenchymal Stem Cells Enhance the Osteoblastic Differentiation of Periodontal Ligament Stem Cells Under high glucose Conditions Through the PI3K/AKT Signaling Pathway

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Biomedical and Environmental Sciences 2022年第9期35卷 811-820页

作者： YANG Shuo ZHU Biao TIAN Xiao Yu YU Han Ying QIAO Bo ZHAO Li Sheng ZHANG Bin Department of Stomatology The First Medical CenterChinese PLA General HospitalBeijing 100853China Department of Stomatology Fuxing HospitalCapital Medical UniversityBeijing 100038China

Objective high glucose(HG)can influence the osteogenic differentiation ability of periodontal ligament stem cells(PDLSCs).Human umbilical cord mesenchymal stem cell-derived exosomes(hUCMSC-exo)have broad application prospects in tissue *** current study aimed to explore whether hUCMSC-exo could promote the osteogenic differentiation of hPDLSCs under HG conditions and the underlying *** We used a 30 mmol/L glucose concentration to simulate HG ***-8 assay was performed to evaluate the effect of hUCMSC-exo on the proliferation of *** phosphatase(ALP)staining,ALP activity,and qRT-PCR were performed to evaluate the pro-osteogenic effect of hUCMSC-exo on *** blot analysis was conducted to evaluate the underlying *** The results of the CCK-8 assay,ALP staining,ALP activity,and qRT-PCR assay showed that hUCMSC-exo significantly promoted cell proliferation and osteogenic differentiation in a dosedependent *** Western blot results revealed that hUCMSC-exo significantly increased the levels of p-PI3K and p-AKT in cells,and the effect was inhibited by LY294002(PI3K inhibitor)or MK2206(AKT inhibitor),***,the increases in osteogenic indicators induced by hUCMSC-exo were significantly suppressed by LY294002 and *** hUCMSC-exo promote the osteogenic differentiation of hPDLSCs under HG conditions through the PI3K/AKT signaling pathway.

关键词： Exosomes Human umbilical cord mesenchymal stem cell Periodontal ligament stem cell Osteogenic differentiation high glucose PI3K/AKT

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Effect of icariin on apoptosis in hippocampal neurons cultured in high glucose

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Journal of Traditional Chinese Medicine 2018年第4期38卷 556-561页

作者： Yan Bin Wang Jingbo Zhang Hong Tian Guoqing Liu Yuqin Department of Traditional Chinese Medicine Peking Union Medical College HospitalChinese Academy of Medical Sciences Institute of Basic Medical Sciences Chinese Academy of Medical Sciences

OBJECTIVE:To investigate effect of icariin on apoptosis in hippocampal neurons cultured in high glucose,and the possible mechanism behind the ***:Hippocampus was obtained from newborn 24 h Sprague-Dawley rats and then primarily *** hippocampal neurons were divided into normal control group,high glucose group,icariin group,icariin + protein kinase B(Akt) inhibitor group and Akt agonist *** each group was cultured in different conditioned medium for 72 h,we detected the apoptosis of neurons with flow cytometry,and the expression of Akt,p-Akt,B-cell lymphoma-2(Bcl-2) and Bcl-2 associated X protein(Bax) with western ***:Compared with the normal control group,the apoptosis rate of hippocampal neurons increased significantly(P high glucose *** with the high glucose group,the apoptosis rate of hippocampal neurons decreased significantly(P high glucose,which may be achieved by increasing the phosphorylation of Akt protein in Akt signal pathway,then increasing the expression of Bcl-2 and inhibiting the expression of Bax.

关键词： Icariin Protein kinase B high glucose Hippocampal neurons Apoptosis

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Inhibition of the negative effect of high glucose on osteogenic differentiation of bone marrow stromal cells by silicon ions from calcium silicate bioceramics

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Regenerative Biomaterials 2020年第1期7卷 9-17页

作者： Xixi Dong Xiaoya Wang Min Xing Cancan Zhao Bin Guo Junkai Cao Jiang Chang Stomatology Department General Hospital of Chinese PLA28 Fu Xing RoadBeijing 100853PR China State Key Laboratory of high Performance Ceramics and Superfine Microstructure Shanghai Institute of CeramicsChinese Academy of Sciences1295 Dingxi RoadShanghai 200050PR China

Human bone marrow stem cells(hBMSCs)are exploited for miscellaneous applications in bone tissue engineering where they are mainly used as seed ***,high glucose(HG)environment has negative impacts on the proliferation and osteogenic differentiation of hBMSCs,thus reducing the bone formation in diabetic *** our former research works,we discovered that silicon(Si)ions extracted from silicate-based bioceramics are able to stimulate the proliferation and osteogenic differentiation of hBMSCs under normal culture *** study aimed to investigate if Si ions could prevent HG-induced inhibition of proliferation and osteogenesis of *** found that 2.59ppm concentration of Si ions promoted the proliferation of hBMSCs under HG *** results from alkaline phosphatase(ALP)activity assay,Alizarin red S staining and quantitative real-time PCR analysis of osteogenic genes(BMP2,RUNX2,ALP,COL1 and OCN)demonstrated that the 15.92ppm concentration of Si ions prevented HG-induced inhibition of the osteogenic differentiation of ***,application of Si ions reduced the level of reactive oxygen species in HG-treated *** HG-treated hBMSCs following 15.92ppm Si ions treatment,activation of BMP2/SMAD signaling pathway was detected,as indicated by the increased expression of BMP2 receptors and its downstream genes such as SMAD1,SMAD4 and *** together,we provide evidence that the specific concentration of Si ions compensated HG-induced inhibition of proliferation and osteogenic differentiation of hBMSCs through antioxidant effect and modulation of BMP2/SMAD *** results suggest that silicate-based bioceramics might be good scaffold biomaterials for bone engineering applications in diabetes patients.

关键词： high glucose silicate osteogenic differentiation bone tissue engineering

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