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Chinese Medical Journal 2003年第4期116卷 554-557页

作者：黄颂敏刘芳沙朝晖付平杨一帆徐勇周海燕 Department of Nephrology West China Hospital of Sichuan University Department of Endocrinology the First Affiliated Hospital of Luzhou Medical College

To observe the effect of high glucose, angiotensin Ⅱ (AngⅡ) and Losartan on the expression of connective tissue growth factor (CTGF) mRNA in cultured mesangial cells (MCs) Methods MCs of SD rats were isolated and cultured high glucose (30 mmol/L) and AngⅡ (10 -9 , 10 - 7 , and 10 -5 mol/L) were added to the medium for 72 hours to observe the influence on CTGF mRNA expression Losartan of 10 -5 mol/L and AngⅡ of 10 -5 mol/L were added to the medium to observe the effects of Losartan on CTGF mRNA expression stimulated by AngⅡ The expressions of CTGF mRNA were detected by reverse transcriptase polymerase chain reaction (RT-PCR) Results RT-PCR showed that high glucose and AngⅡ up-regulated the expression of CTGF mRNA, and AngⅡ stimulated the expression in a dose-dependent manner Expression of CTGF mRNA induced by AngⅡwas partially suppressed by 10 -5 mol/L Losartan (Phigh glucose and AngⅡ can enhance the expression of CTGF mRNA and thus be involved in the process of renal fibrosis Losartan can have a partial fibrogenesis-inhibiting effect, with implications for the treatment of renal fibrosis

关键词： high glucose angiotensin Ⅱ Losartan connective tissue growth factor ·mesangial cells renal fibrosis

Effect of Chinese Herbal Compound Naofucong (脑复聪) on theInflammatory Process Induced by high glucose in BV-2 Cells

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Chinese Journal of Integrative Medicine 2016年第11期22卷 832-839页

作者：景光婵张孟仁纪超左萍萍刘玉琴顾蓓 Department of Traditional Chinese Medicine Peking Union Medical College Hospital Peking Union Medical College and Chinese Academy of Medical Sciences Beijing 100730 China Institute of Basic Medical Sciences Chinese Academy of Medical Sciences and School of Basic Medicine Peking Union Medical College Beijing 100005 China

Objective: To determine the effect of medicated serum of Chinese herbal compound Naofucong(脑复聪, NFC) on the microglia BV-2 cells viability and the transcription and expression of interleukin-6(IL-6) and tumor necrosis factor α(TNF-α) in microglia BV-2 cells to further explore the mechanisms underlying the protective effect of NFC on inflammatory process induced by high glucose. Methods: The microglia BV-2 cells incubated in vitrowere divided into different groups: the control group(25 mmol/L glucose), the model group(75 mmol/L glucose), high glucose media containing different dose medicated serum of NFC. After being cultured for 24 h, changes in IL-6 and TNF-α were measured by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay. The expression of surface marker CD11 b of activated microglia was measured by confocal laser scanning microscope and Western blot. Nuclear factor-κB(NF-κB) p-p65 expression was analyzed by Western blot. Results: The model group obviously increased the expression of microglial surface marker CD11 b and NF-κB p-p65(all Phigh glucose for 24 h. Conclusions: The inhibition of microglial activation and IL-6 and TNF-α expression induced by high glucose may at least partly explain NFC therapeutic effects on diabetes-associated cognitive decline diseases. Its underlying mechanism could probably be related to the inhibition of NFC on NF-κB phosphorylation.

关键词： high glucose microglia inflammatory cytokine Chinese herbal compound Naofucong

Effect of Serum Containing Jinmaitong Capsule(筋脉通胶囊)on Rats' Schwann Cell Apoptosis Induced by high glucose Concentration

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Chinese Journal of Integrative Medicine 2013年第7期19卷 517-523页

作者：王普艳梁晓春张宏赵丽孙青黄文智李伯武 Department of Traditional Chinese Medicine Peking Union Medical College HospitalChinese Academy of Medical Sciences Department of Traditional Chinese Medicine 306th Hospital of People's Liberation Army Institute of Basic Medical Science Peking Union Medical CollegeChinese Academy of Medical Sciences

To evaluate the effect of serum containing Jinmaitong Capsule （筋脉通胶囊, JMT） on apoptosis of Schwann cells （SCs） that are cultured in high glucose at the cellular and molecular levels. Methods： SCs were cultured in Dulbecco＇s modified Eagle＇s medium （control group）, high glucose （50 mmol/L） medium supplemented with 20% rat serum （HG group）, and 50 mmol/L glucose medium supplemented with serum containing JMT （JMT group）. SC apoptosis was detected using a terminal deoxynucleotidyl transferase dUTP nick end labeling kit. The expression of Bcl-2 and the caspase-3 p20 subunit in SCs were detected by real- time fluorogenic quantitative polymerase chain reaction and confocal laser scanning microscopy, respectively. Results： No apoptosis was detected in SCs that were cultured in the control group. The percentage of apoptosis of SCs cultured in the HG group was much higher than that in the control group. The apoptosis of SCs in the JMT group was lower than that in the HG group. Fluorescence intensity of Bcl-2 and the expression of Bcl-2 mRNA in SCs that were cultured in the HG group were much lower than those in the control group and much higher than those in the JMT group （P〈0.01）. The fluorescence intensity of caspase-3 p20 and the expression of caspase-3 p20 mRNA in SCs that were cultured in the HG group were much higher than those in the control group （P〈0.01）, and they were remarkably lower in the JMT group （P〈0.01）. Conclusions： JMT effectively prevents SC apoptosis that is induced by high glucose. This effect may be because of increased expression of Bcl-2 mRNA and protein and decreased expression of caspase-3 p20 mRNA and protein.

关键词： Jinmaitong Capsule apoptosis high glucose schwann cells

high glucose alters HEK293 cell growth by inhibition of cloned BKCa channel

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High glucose alters HEK293 cell growth by inhibition of clon...

中国生理学会第23届全国会员代表大会暨生理学学术大会

作者： Man-Jiang XIE,Hui CHANG,Yu-Guang MA,Zhen SONG,Jin MA,Zhi-Bin YU Department of Aerospace Physiology,Key Laboratory of Aerospace Medicine of Ministry of Education,Fourth Military Medical University,Xi’an 710032,Shaanxi Province,China

It has been reported that diabetic vascular dysfunction is associated with impaired function of large conductance Ca-activated K(BK) channels which are key determinants in ***,it is unclear whether impaired BK channel directly participates in regulating diabetic vascular remodeling by altering cell growth in response to *** present study,we investigated the specific role of BK channel in controlling cell growth(apoptosis and proliferation) under high glucose concentration(25 mmol/L).The cDNA encoding theα+β1 subunit of BK channel,hSloα+β1,was transiently transfected into human embryonic kidney 293(HEK293) cells. Cloned BK currents were recorded by both whole-cell and cell-attached patch clamp *** apoptosis was assessed with immunocytochemistry and analysis of fragmented DNA by agarose gel *** proliferation was investigated by flow cytometry assays,MTT test,and *** addition,the expression of anti-apoptotic protein Bcl-2,intracellular Ca,and mitochondrial membrane potential(△ψm) were also examined to investigate the possible *** results indicate that inhibited cloned BK channels might be responsible for hyperglycemia-altered HEK-hSloα+β1 cell ***,activation of cloned BK channel by NS1619 or Tamoxifen,the agonists of BK channel significantly induced apoptosis and suppressed high glucose-induced proliferation in EEK-hSloα+β1 cells under hyperglycemia ***,the possible mechanisms of activation of BK channel under high glucose condition were associated with decreased expression of Bcl-2,elevation of intracellular Ca,and a concomitant depolarization of△ψm in HEK-hSloα+β1 *** conclusion, cloned BK channel directly regulated HEK293cell growth in response to high glucose.

关键词： large conductance Ca2+-activated potassium(BKCa) channel apoptosis proliferation hSloα+β1 human embryonic kidney 293(HEK293) cells high glucose

Effect of Jinmaitong (筋脉通) Serum on the Proliferation of Rat Schwann Cells Cultured in high glucose Medium

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Chinese Journal of Integrative Medicine 2008年第4期14卷 293-297页

作者：屈岭梁晓春张宏吴群励孙连庆顾蓓 Department of Traditional Chinese Medicine Peking Union Medical College HospitalChinese Academy of Medical Sciences Cell Center Institute of Basic MedicalSciencePeking Union Medical CollegeChinese Academy of Medical Sciences Cell Center Institute of Basic MedicaISciencePeking Union Medical CollegeChinese Academy of Medical Sciences

Objective： To investigate the effect of Jinmaitong （筋脉通,JMT） serum on the proliferation of rat Schwann cells （SCs） primarily cultured in high glucose medium. Method： SOs were primarily cultured in Dulbecco＇s minmum essential medium （DMEM control）, 50 mmol/L glucose medium （50 mmol/L Glu）, 75 mmol/L glucose medium （75 mmol/L Glu）, as well as 50 mmol/L glucose medium, with different concentrations of JMT serum （undiluted, 1：2 diluted and 1：8 diluted） and Neurotropin （Ntp）, respectively. The proliferation of SCs under different conditions was detected by MTT. Result： SCs grew exuberantly in DMEM within 24-72 h, but slowed down at 96 h. The proliferation of SCs was inhibited in 50 mmol/L Glu and 75 mmol/L Glu after cultures of 48, 72 and 96 h, which showed that both were significantly different compared to the control group （P〈0.01）. The inhibition was more significant in 75 mmol/L Glu than in 50 mmol/L Glu （P〈0.05）. Spearman＇s rho analysis revealed that the proliferation of SCs had a negative correlation with the concentration of glucose （r=-0.471, P〈0.01）. Excluding the time factor, partial correlation showed similar results （r =-0.679, P〈0.01）. After 48 h, the proliferation of SCs increased significantly in JMT 1：2 and Ntp compared with 50 mmol/L Glu （control 0.437±0.019, 50 mmol/ L Glu 0.367±0.035, JMT1：2 0.426±0.024, Ntp 0.422±0.013; P〈0.01）, and there were no statistically significant differences among the JMT groups, the Ntp group and the control group （P〉0.05）. Conclusions： The proliferation of SCs was inhibited in high glucose medium, and the inhibition was reduced by different concentrations of JMT serum, especially at JMT1：2.

关键词： Schwann cells high glucose primary culture Jinmaitong proliferation Chinese herbs' serum pharmacology

Neuroprotective Effect of Bu-Shen-Huo-Xue Extract against high glucose-induced Apoptosis in PC12 Cells

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TMR Modern Herbal Medicine 2018年第3期1卷 143-154页

作者： Shao-Yang Zhao Xin Dong Peng-Fei Tu Ke-Wu Zeng Xue-Mei Wang Research Studio of Integration of Traditional and Western Medicine First Hospital Peking University Beijing China State Key Laboratory of Natural and Biomimetic Drugs School of Pharmaceutical Sciences Peking University Beijing China

Objective： To investigate the neuroprotective effect of Bu-Shen-Huo-Xue （BSHX） extract, a polyherbal formula, against high glucose （HG）-induced neurotoxicity in PC12 cells. Methods： Cell viability assay, Lactate Dehydrogenase （LDH） assay, Reactive Oxygen Species （ROS） detection, Hoechst 33258, Acridine Orange （AO）/Ethidium Bromide （EB） double stain and Mitochondrial Membrane Potential （MMP） assay were performed. In addition, Bax, Bcl-2, caspase-3, cleaved caspase-3, PARP, cleaved PARP, cytochrome c and Mitogen-Activated Protein Kinases （MAPKs） were detected by western blot. Results： BSHX extract increased cell viability and decreased LDH leakage in a concentration-dependent manner in HG-induced PC12 cells. Moreover, BSHX extract decreased the level of intracellular ROS, increased mitochondrial membrane potential, regulated the expressions of Bax and Bcl-2, and inhibited the release of cytochrome c from mitochondria. Furthermore, BSHX extract attenuated the activation of caspase-3 and PARP, and inhibited the phosphorylations of c-Jun N-terminal kinase （JNK） and p38 MAPKs. Conclusion： BSHX extract exhibited significant neuroprotective effect on HG-induced apoptosis in PC12 cells. This effect may be associated with the suppression of ROS generation as well as mitochondria-mediated caspase and JNK/p38 MAPK signaling pathways.

关键词： diabetic encephalopathy traditional Chinese medicine Bu-Shen-Huo-Xue extract neuroprotective high glucose

Pterostilbene attenuates high glucose-induced oxidative injury in hippocampal neuronal cells by activating nuclear factor erythroid 2-related factor 2

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Pterostilbene attenuates high glucose-induced oxidative inju...

第十三届中国医师协会神经外科医师年会

作者： Yang Yang Chongxi Fan Bodong Wang Zhiqiang Ma Dongjin Wang Bing Gong Shouyin Di Shuai Jiang Yue Li Tian Li Zhi Yang Erping Luo Department of Biomedical Engineering The Fourth Military Medical University Department of Thoracic and Cardiovascular Surgery Affiliated Drum Tower Hospital of Nanjing University Medical School Department of Thoracic Surgery Tangdu HospitalThe Fourth Military Medical University Graduate Management Team The Fourth Military Medical University Department of Aerospace Medicine The Fourth Military Medical University

In the present study,neuroblastoma（SH-SY5 Y）cells were used to investigate the mechanisms mediating the potential protective effects of pterostilbene（PTE）against mitochondrial metabolic impairment and oxidative stress induced by hyperglycemia for mimicking the diabetic *** glucose medium（100 mM）decreased cellular viability after 24 h incubation which was evidenced by:（i）reduced mitochondrial complex I and III activities;（ii）reduced mitochondrial cytochrome C;（iii）increased reactive oxygen species（ROS）generation;（iv）decreased mitochondrial membrane potential（ΔΨm）;and（v）increased lactate dehydrogenase（LDH）***（2.5,5,and 10 μM for 24 h）was nontoxic and induced the nuclear transition of *** of PTE（2.5,5,and 10 μM for 2 h）displayed a dose-dependently neuroprotective effect,as indicated by significantly prevented high glucose-induced loss of cellular viability,generation of ROS,reduced mitochondrial complex I and III activities,reduced mitochondrial cytochrome C,decreased ΔΨm,and increased LDH ***,the levels of nuclear factor erythroid 2-related factor 2（Nrf2）,heme oxygenase-1（HO-1）and glutathione S-transferase（GST）were elevated after PTE *** addition,the elevation of nuclear Nrf2 by PTE treatment（10 μM for 2 h）was abolished by Nrf2 ***,Nrf2 siRNA induced the opposite changes in mitochondrial complex I and III activities,mitochondrial cytochrome C,reactive species generation,ΔΨm,and ***,the present findings were the first to show that pterostilbene attenuated high glucose-induced central nervous system injury in vitro through the activation of Nrf2 signaling,displaying protective effects against mitochondrial dysfunction-derived oxidative stress.

关键词： high glucose Oxidative stress Pterostilbene Nuclear factor erythroid 2-related factor 2 signaling Neuroprotection

Paeonol blocks Ras-Raf-ERK1/2 signal pathway in rat vascular smooth muscle cells co-cultured with high glucose stimulated vascular endothelial cells

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Paeonol blocks Ras-Raf-ERK1/2 signal pathway in rat vascular...

安徽省药理学会2012年学术年会

作者： Jun-jun Chen Min Dai Yue-qin Wang Key Laboratory of Xin’an Medicine Ministry of EducationAnhui Province Key Laboratory of R&D of Chinese Medicine

Aim Paeonol(Pae) has been previously reported to against artheroclerosis via inhibiting vasular smooth muscle cell(VSMC) proliferation or vascular endothelial cell(VEC) inflammatory injury *** current study was based on co-cultured system of VSMCs and VECs to investigate the protective mechanism of Pae on atherosclerosis(AS) by researching regulation of secretory function of VECs and proliferation of VSMCs through Ras-Raf-ERK1/2 signal pathway. Methods The co-culture model of rat VEC and VSMC was set up with transwell culture *** were stimulated by high glucose to further explore the mechanism of diabetes-induced vascular disease and intervention of *** cells survival was detected by MTT and LDH kit to measure the damage of *** was also used to measure the proliferation in *** of VEGF was detected by *** levels of Ras,Raf,P-Raf,ERK1/2,and P-ERK1/2 were determined by western *** We found that high concentration(35.5μM) of glucose induced in 48h damaged VECs in greatest *** injury of VECs stimulated VSMC proliferation in co-cultured *** could inhibit VSMC proliferation via decreasing platelet derivative growth factor(PDGF) release in VECs and inhibiting over-expression of protein Ras,P-Raf and P-ERK in *** in concentrtion of 120μM showed the greatest *** The results indicated that diabete modulated inflammatory factor expression in *** stimulated VSMC proliferation and promoted the development of *** exerted inhibitory effect on VSMC *** inhibitory mechanism of Pae may be realized by inhibiting the secretion of PDGF in VECs,thus inhibiting Ras-Raf-ERK1/2 signal pathway in ***,Pae may effect on Ras-Raf-ERK1/2 signal pathway *** experiment would provide theoretical basis for Pae in clinical treatment on AS and build a technical platform for new drug’s targets.

关键词： Paeonol high glucose Ras-Raf-ERK1/2 signal pathway vascular endothelial cells vascular smooth muscle cells co-cultured system

The research on Toll-like receptors signaling pathway induced by high glucose in gingival fibroblasts

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The research on Toll-like receptors signaling pathway induce...

中华口腔医学会第14次全国口腔医学学术会议（2012年会）

作者：尚婷婷天津医科大学口腔

Abstract Objective:The study is to explore the singnaling pathway of Toll-like receptors(TLRs)2 and 4 induced by high glucose in human gingival fibroblasts(HGFs)with small interfering RNA(siRNA)and real-time PCR,which provides theoretical basis for elucidating pathogenesis of periodontitis combined with diabetes and novel ideas for treatment of periodontitis. Method:After gingival tissues were digested by DispaseⅡand removed gingival epithelium,HGFs were obtained from the primary culture of *** chemical synthesized siRNAs labeling fluorescence were transfected into HGFs with ROCHE transfection *** concentration(30 nM,50 nM and 100 nM) and time(6h,12h,24h,36h,48h,60h and 72h)were set up to determine the best transefection efficiency through observing *** expressions of TLR2 and TLR4 were detected by real-time PCR under the stimulation of high *** PKC-a siRNA,PKC-P siRNA and PKC-5 siRNA were applied to block PKC *** expressionss of TLRs,NF-kB p50 and NF-kB p65 were detected by real-time PCR after the stimulation of high glucose in comparison to the low glucose culture. ResultrHGFs were successfully cultured by tissue-explant *** optimum transfection efficiency was over 95%when the transfection concentration was 50 nM and time was 48 *** mRNA level of TLR2 in HGFs under the stimulation of high glucose significantly increased(p0.05).PKC-a siRNA and PKC-5 siRNA down-regulated mRNA levels of TLR2 and NF-kB p65(phigh glucose,however,the PKC-P siRNA showed no effect on the mRNA levels of TLR2 and NF-kB p65(p>0.05).There was no change in the level of NF-kB p50(p>0.05)under the stimulation of high glucose compared to the low glucose ***,PKC-a siRNAs PKC-P siRNA and PKC-5 siRNA did not change the level of NF-kB p50 in HGFs. Conclusion:HGFs can be successfully cultured by tissue-explant *** optimum transfection e

关键词： human gingival fibroblasts high glucose siRNA Toll-like receptors signaling pathway

Effect of high glucose on cytokines secretion in isolated perfused rat kidney and rat mesangial cells

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Effect of high glucose on cytokines secretion in isolated pe...

中国药理学会第十次全国学术会议

作者： Ya-Jie Hu,Bo Jiao (Department of Pharmacology,School of Pharmaceutical Sciences,Shandong University,Jinan 250012,China)

Objective To investigate the expression of TGF-β1,VEGF and PDGF-BB in response to high glucose in isolated perfused kidney(IPK) and cultured mesangial cells(MCs).Methods The IPK and cultured MCs were used as *** IPK model,the urine and perfusate samples were collected at 0.5h,1 h,2h,4h and 6h after *** urine?ow,creatinine clearance rate,activity of LDH and urine kalium excretion was detected to assess kidney *** MCs were incubated for 0.5h,1.0h,2.0h,4.0h and *** cytokines expression was measured by immunohistochemistry and *** The expression of TGF-β1 and PDGF-BB was increased,and VEGF was increased before 2h and decreased at 4h and 6h compared with normal in IPK ***-β1 was lower at 4h than that at 4h and *** MCs,the expression of TGF-β1 and VEGF was the same as that in IPK model,and PDGF-BB was increased ***:The expression of the three cytokines stimulated by high glucose was changed in the IPK and cultured mesangial *** there were interaction between them.

关键词： isolated perfused kidney mesangial cells high glucose TGF-β1 VEGF PDGF-BB