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THE EXPERIMENTAL STUDY ON THE CELL APOPTOSIS AND EXPRESSION OF bcl-2 protein IN INTRACEREBRAL HEMORRHAGE IN MODEL OF RATS

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Journal of Pharmaceutical Analysis 2006年第1期18卷 61-64页

作者：鲍刚郭宁张仲林陈伟鲍得虎 Department of Neurosurgery the First Affiliated Hospital of Medical SchoolXi'an Jiaotong UniversityXi'an 710061China Department of Cardiology the First Affiliated Hospital of Medical SchoolXian Jiaotong UniversityXian 710061China

Objective To study whether there is the apoptosis of neural cells and the expression of bcl-2 protein in intracerebral hemorrhage (ICH) in model of rats, for the further understanding the mechanism of the delayed damage of the neural cells around the hematoma after ICH. Methods Fifty SD rats were randomly divided into 5 groups, ten in each. With the Group A as the control, the rest 40 were used to set up intracerebral hemorrhage model. The brains were taken out at 12 th , 24 th , 48 th and 72 th hours, respectively. Apoptosis cells were detected with terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL), and the expression of bcl-2 protein was detected with immunochemical stainging methed (SP). Results In the control group, no apoptosis cells and Bc1-2 protein were detected. In rest groups, the apoptosis cells and Bc1-2 protein were expressed in different degree. Apoptosis rates verified and corresponded with the time after ICH, with the peak at 48 th -72 th hour after hemorrhage. The peak rate of apoptosis cells was (24.50±2.69)% and bcl-2 protein expression was (20.76±1.97)% . There was significant difference between the experimental groups and control (Pbcl-2 protein. Conclusion Apoptosis may be an important factor in the secondary trauma of ICH. There is a time leg after hemorrhage. All this is instructive to clinical treatment in time. bcl-2 protein keeps increasing in a certain time after hemorrhage, but not synchronize with the cell apoptosis. This indicates that bcl-2 has the effect to reduce the apoptosis of neural cells.

关键词： apoptosis intracerebral hemorrhage bcl-2 protein rat

Pretreatment with low-frequency repetitive transcranial magnetic stimulation may influence neuronal bcl-2 and Fas protein expression in the CA1 region of the hippocampus: A possible anti-epilepsy mechanism in a lithium-pilocarpine-induced epileptic rat mod

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Neural Regeneration Research 2010年第12期5卷 895-900页

作者： Sha Ke Hongning Zhao Xiaoming Wang Junqiang Zhang Fang Chen Yuanxu Wang Xiaoqiong Zhao Hui Huang Jianxiu Hu Department of Electroneurophysiology Affiliated Hospital of North Sichuan Medical CollegeNanchong 637000 Sichuan Province China Department of Electroneurophysiology Research Institute of Neurological Disease North Sichuan Medical College Nanchong 637000 Sichuan Province China

BAOKGROUND： bcl-2 and Fas proteins are well known as anti-apoptotic and pro-apoptotic factors respectively. However, whether the anti-epileptic mechanism of low-frequency repetitive transcranial magnetic stimulation （rTMS） involves an anti-apoptotic effect via regulating bcl-2 and Fas protein expression remains to be determined. OBJECTIVE： To verify the correlation between the anti-epileptic mechanism following pretreatment of low-frequency rTMS and anti-hippocampal apoptosis. DESIGN, TIME AND SETTING： A randomized controlled animal experiment was performed at Institute of Neurological Disorders, Affiliated Hospital of North Sichuan Medical College between September 2007 and March 2008. MATERIALS： Pilocarpine （053K13011） was provided by Sigma, USA; lithium was provided by Shanghai Biotechnology Co., Ltd., China; Dantec Maglite-r25 rTMS instrument was provided by Dundee, Denmark. METHODS： A total of 21 adult male Wistar rats were randomly divided into control （n = 6）, rTMS pretreatment （n = 9）, and sham-stimulation （n = 6） groups. The rTMS pretreatment group was pretreated with low-frequency rTMS （0.5 Hz, 75% threshold intensity, 20 times/bundle, and 5 bundles/day）, while the sham-stimulation group was sham-stimulated with a similar sound for 7 successive days to establish lithium-pilocarpine-induced epileptic state models. MAIN OUTCOME MEASURES： Epileptic stroke latency; neuronal morphology was observed using hematoxylin and eosin staining; mean positive-reactive cell number and mean absorbance of bcl-2 and Fas protein in the hippocampal CA1 region was observed using immunohistochemistry. RESULTS： Epileptic latency in the rTMS pretreatment group was significantly enhanced （P 〈 0.01）, and a number of degenerated neurons were observed to be apoptotic. bcl-2 protein expression increased at each time point, but Fas protein expression decreased （P 〈 0.01）. CONCLUSION： Low-frequency rTMS has an anti-epileptic effect, which may be via regulation of bcl-2 and Fas protein ex

关键词： transcranial magnetic stimulation epileptic state bcl-2 protein Fas protein brain injury neural regeneration

The Expression of Apoptosis-Related Genes bcl-2 and Bax protein and Apoptosis Positivity in Cervical Carcinoma during Irradiation

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The Chinese-German Journal of Clinical Oncology 2005年第2期4卷 105-107页

作者：赵东利石景森李明众宋丽萍王书文西安交通大学第一医院肿瘤放疗科 710061

To evaluate the apoptosis positivity, the expression of bcl-2, bax proteinsin 30 patients with squamous cell cervix carcinoma before and after radiotherapy. Methods: By usingimmuno-histochemical and TDT-dUTP nick end labelling techniques, 30 cases of squamous cell cervicalcarcinoma were analyzed. Results: The apoptosis positivity before and after irradiation was 76.7%and 100% respectively, with the difference being significant (P bcl-2protein before and after irradiation were 73.3% and 46.7% respectively, with the difference beingsignificant (P protein before and after irradiation were 86% and100% respectively, with the difference being significant (P bcl-2protein play an important role in apoptosis induced by fractionated radiation therapy. Apoptosisinduced by irradiation is contributed to upregulation of bax protein or downregulation of bcl-2protein.

关键词： cervical carcinoma radiotherapy apoptosis positivity bcl-2 protein baxprotein

THE QUANTITATIVE MEASUREMENT OF bcl-2, P53 protein AND PCNA EXPRESSION IN BREAST CARCINOMA AND THEIR CORRELATION WITH PROGNOSIS

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Journal of Pharmaceutical Analysis 1998年第2期12卷 120-124,132页

作者：张学斌王鸿雁 Department of Pathology First Affiliated Hospital Xi'an 710061 China

To study quantitative index of bci-2, P53, Nroliferating cell nuclear antigen (PCNA),ER and PR in breast carcinoma and their correiation and their relatiousbip with prognosis, the ex expression of bcl-2, P53 and PCNA were studied by immunohistochemical technique. The measurementof ER and PR used enzyme linked affinuity histochemical methods. The quantitative index was analyzed by image technique. All analyses were hased on 60 breast carcinomas. The results were as follows:the more bcl-2 protein, the lower histological graded the longer survival term and the highersurvival rate (Pbcl-2, P53 and PCNA expression were ofvalue in evaluating the degree of differentiation and prognosis in breast carcinoma. The quantitativeand qualitative measurement or p53 protein expression showed a Ⅰwerful evidence in evaluatingprognosis of bcl-2 were more significant in evaluating poor prognosis of breast carcinoma. A relationship between bcl-2 and ER, PR showed a better value for response to endocrine therapy in breastcarcinoma patients.

关键词： breast carcinoma P53 protein bcl-2 protein PCNA image analysis technique

Specific COX-2 inhibitor NS398 induces apoptosis in human liver cancer cell line HepG2 through bcl-2

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World Journal of Gastroenterology 2005年第2期11卷 204-207页

作者： Dong-ShengHuang Ke-ZhenShen Jian-FengWei Thng-BoLiang Shu-SenZheng Hai-YangXie DepartmentofHepatobiliaryPancreaticSurgery TheFirstAffiliatedHospitalZhejiangUniversityCollegeofMedicineHangzhou310003ZhejiangProvinceChina

AIM: To evaluate the effects of NS-398, a cyclooxygenase-2 (COX-2) inhibitor, on the proliferation and apoptosis of HepG2 cells. METHODS: The effects of NS-398 on the proliferation of HepG2 cells were evaluated by MTT. DNA fragmentation gel analysis was used to analyze the apoptotic cells. DNA ploidy and apoptotic cell percentage were calculated by flow cytornetry. The expression of COX-2 and bcl-2 mRNA was identified by competitive RT-PCR. Furthermore, expression level of bcl-2 was detected using Western blot in HepG2 after treated with NS-398. RESULTS: NS-398 inhibited cell proliferation and induced apoptosis of HepG2 cells in a concentration-dependent manner. DNA ploidy analysis showed that S phase cells were significantly decreased with increase of NS-398 concentration. The quiescent GO/G1 phase was accumulated with decrease of bcl-2 mRNA. Whereas NS-398 had no effect on the expression of COX-2 mRNA, and no correlations were found between COX-2 mRNA and HepG2 cell proliferation and apoptosis induced by NS-398 (r=0.056 and r=0.119, respectively). bcl-2 protein level was inhibited after treated with NS-398. CONCLUSION: NS-398 significantly inhibits the proliferation and induces apoptosis of HepG2 cells. Mechanisms involved may be accumulation of quiescent GO/G1 phase and decrease of bcl-2 expression.

关键词： Liver cancer NS-398 bcl-2 protein COX-2

Effects of Ethyl Pyruvate on Myocardial Apoptosis and Expression of bcl-2 and Bax proteins after Ischemia-reperfusion in Rats

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2008年第3期28卷 281-283页

作者：郭家龙张凯伦季艳梅蒋雄刚左顺庆 Department of Cardiovascular Surgery Union HospitalTongji Medical CollegeHuazhong University of Science and Technology Department of Intensive Care Unit Taihe Hospital Affiliated to Yunyang Medical College Department of Thoracic Surgery Taihe Hospital Affiliated to Yunyang Medical College

In order to study the effects of ethyl pyruvate on cardiomyocyte apoptosis following ischemia/reperfusion （I/R） in vitro and the expression of bcl-2 and Bax proteins, isolated rat hearts were perfused in a Langendorff model. Twenty-four rats were randomly divided into 3 groups （n=8 in each group）： control group was perfused for 120 min. In the I/R group, after 30 min stabilization the injury was induced by 30 min global ischemia followed by 60 min reperfusion. Ethyl pyruvate （EP） group was set up with the same protocol as I/R group except that it was supplied with 2 mmol/L EP 15 rain before ischemia and throughout reperfusion. Myocardial malonaldehyde （MDA） content was measured. Myocardial apoptotic index （AI） was tested by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling （TUNEL） method. The expression of anti-apoptotic protein bcl-2 and pro-apoptotic protein Bax in cardiac myocytes was detected by immunohistochemistry. As compared with control group, the content of MDA, myocardial AI and the expression of bcl-2, Bax proteins were increased significantly in I/R group, but the content of MDA, myocardial AI and the expression of Bax protein were decreased obviously and the expression of bcl-2 protein was up-regulated in EP group （P〈0.05）. These results demonstrate that EP could inhibit apoptosis of cardiac myocytes possibly via alleviating oxidative stress, up-regulating bcl-2 and down-regulating Bax proteins.

关键词： ethyl pyruvate myocardial reperfusion injury apoptosis bcl-2 protein Bax protein

Abrogation of heat-shock protein (HSP)70 expression induced cell growth inhibition and apoptosis in human androgen-independent prostate cancer cell line PC-3m

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Asian Journal of Andrology 2004年第4期6卷 319-324页

作者： Zhi-GangZhao Qing-ZhengMa Chun-XiaoXu DepartmentofUrology ShantouUniversityMedicalCollegeShantou515031China DepartmentofUrology ShandongUniversityClinicalMedicalCollegeShandongProvinceHospitalJinan250021China DepartmentofUrology thesecondAffiliatedHospitalShandongUniversityMedicalCollegeJinan250012China

Aim: To investigate the effect of abrogating heat shock protein (HSP) 70 expression by antisense HSP70 oligonucleotides treatment on human androgen-independent prostate cancer cell line PC-3m growth. Methods: PC-3m cells were treated with 0-16 μmol/L antisense HSP70 oligomers for 0-100 hr. Cell growth inhibition was analyzed using a trypan blue dye exclusion test. Apoptotic cells were detected and confirmed by flow cytometric analysis and DNA fragmentation analysis. The protein expression of HSP70 and bcl-2 affected by antisense HSP70 oligomers were determined using Western blot. Results: Antisense HSP70 oligomer induced apoptosis and then inhibited proliferation of PC-3m cells in a dose- and time-dependent manner. Ladder-like patterns of DNA fragments were observed in PC-3m cells treated with 10 μmol/L antisense HSP70 oligomer for 48 hr or 8 μmol/L for 72 hr on agarose gel electrophoresis. Antisense HSP70 oligomer pretreatment enhanced the subsequent induction of apoptosis by heat shock in PC-3m cells. In addition, undetectable HSP70 expression was observed at a concentration of 10 μmol/L antisense HSP70 oligomer treatment for 48 hr or 8 μmol/L for 72 hr in Western blot, which was paralleled by decreased expression levels of anti-apoptotic protein bcl-2. Conclusion: HSP70 antisense oligomer treatment abrogates the expression of HSP70, which may disrupt HSP70-bcl-2-interactions and further down-regulate bcl-2 expression, in turn inducing apoptosis and inhibiting cell growth in PC-3m cells.

关键词： prostate carcinoma heat shock protein (HSP) bcl-2 protein apoptosis proliferation

Total flavonoids of hawthorn leaves promote motor function recovery via inhibition of apoptosis after spinal cord injury

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Neural Regeneration Research 2021年第2期16卷 350-356页

作者： Qiong Zhang Yin Xiong Bo Li Gui-Ying Deng Wen-Wen Fu Bai-Chuan Cao Shao-Hui Zong Gao-Feng Zeng School of Public Health Guangxi Medical University NanningGuangxi Zhuang Autonomous RegionChina Department of Spine Osteopathic Surgery The First Affiliated Hospital of Guangxi Medical UniversityGuangxi Zhuang Autonomous RegionChina Research Center for Regenerative Medicine Guangxi Key Laboratory of Regenerative MedicineGuangxi Medical UniversityNanningGuangxi Zhuang Autonomous RegionChina

Flavonoids have been reported to have therapeutic potential for spinal cord *** leaves have abundant content and species of total flavonoids,and studies of the effects of the total flavonoids of hawthorn leaves on spinal cord injury have not been published in or outside ***,Sprague-Dawley rats were used to establish a spinal cord injury model by Allen's *** were intraperitoneally injected with 0.2 m L of different concentrations of total flavonoids of hawthorn leaves(5,10,and 20 mg/kg)after spinal cord *** were administered once every 6 hours,three times a day,for 14 *** treatment with various concentrations of total flavonoids of hawthorn leaves,the Basso,Beattie,and Bresnahan scores and histological staining indicated decreases in the lesion cavity and number of apoptotic cells of the injured spinal cord tissue;the morphological arrangement of the myelin sheath and nerve cells tended to be regular;and the Nissl bodies in neurons *** Basso,Beattie,and Bresnahan scores of treated spinal cord injury rats were *** blot assays showed that the expression levels of pro-apoptotic Bax and cleaved caspase-3 were decreased,but the expression level of the anti-apoptotic bcl-2 protein was *** improvement of the above physiological indicators showed a dose-dependent relationship with the concentration of total flavonoids of hawthorn *** above findings confirm that total flavonoids of hawthorn leaves can reduce apoptosis and exert neuroprotective effects to promote the recovery of the motor function of rats with spinal cord *** study was approved by the Ethics Committee of the Guangxi Medical University of China(approval No.201810042)in October 2018.

关键词： apoptosis Bax protein bcl-2 protein cleaved caspase-3 inflammation motor function recovery neuroprotection Nissl bodies spinal cord injury total flavonoids of hawthorn leaves

Antitumor activity of a novel bis-aziridinylnaphthoquinone (AZ4) mediating cell cycle arrest and apoptosis in non-small cell lung cancer cell line NCl-H460

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Acta Pharmacologica Sinica 2007年第4期28卷 559-566页

作者： Kou-gea SHYU Hsien-shou KUO Yuh-ling LIN Division of Cardiac Shin Kong Wu Ho-Su Memorial HospitalTaipei 106TaiwanChina Department of Biochemistry Taipei Medical UniversityTaipei City 110TaiwanChina Department of Medicine Fu-Jen Catholic UniversityHsin-ChuangTaipei Hsien 242TaiwanChina

Aim:The cytotoxic activities of a series of bis-aziridinylnaphthoquinone,AZ1 toAZ4,on human lung carcinoma cell lines,H460,and normal lung cells fibroblastcell line,MRC-5,and the mechanisms of H460 cells induced by AZ4 ***:The MTT assay was used to determine the *** cycle was analysed by *** activity of caspase 3,8 and 9was determined by cell-permeable fluorogenic detection *** blotassay was used to evaluate the regulation of cyclin B,Cdc-2,p53,p21,and the bcl-2 ***:AZ1 to AZ4 displayed various cytotoxicity activities againstH460 and MRC-5 *** to those compounds,AZ4 was with the mosteffective agent among the 5 tested analogues at reducing H460 cell viability with anIC50value of 1.23 μmol/L;it also exhibited weak cytotoxicity against MRC-5 cells withan IC50value of 12.7 μmol/*** results show that growth arrest on the G2-M phaseof H460 cells induced by AZ4 for 24 h was discovered,and this might be alteredwith the reduced Cdc-2 protein expression of 47% at 2.0 μmol/L AZ4,but not withcyclin B protein *** AZ4 treated cells were then led to apoptosis after48 *** was associated with the activation of apoptotic enzyme caspase 3 andmediated by caspase 8,but not caspase 9 at various concentrations of AZ4 afterbeing cultured for 48 h and 30 h,*** anti-apoptotic protein （bcl-2）expression in H460 cells altered by 39% with downregulation,and the p53 proteinby 25% with upregulation after being cultured with 2.0 μmol/L AZ4 for 48 *** atime-dependent wanrer,the expression of the p53 and p21 proteins were increasedto the maximum at 24 h,and then decreased at ***:AZ4 represents anovel antitumor aziridinylnaphthoquinone with therapeutic potential against thenon-small cell lung cancer cells.

关键词： cell cycle apoptosis bcl-2 protein aziridinylnaphthoquinone