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低氧诱导大鼠外周血单核细胞表达TNF-α与PKC膜易位及活性变化关系的研究 被引量:2
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作者 徐智 吴国明 +4 位作者 钱桂生 王兴胜 陈维中 薛桥 王士雯 《重庆医学》 CAS CSCD 2008年第23期2716-2719,共4页
目的研究低氧诱导大鼠外周血单核细胞(peripheral blood monouclear cells,PBMCs)膜内PKC膜易位及活性变化对肿瘤坏死因子α(TNF-α)表达的作用,探讨缺氧与全身炎症反应综合征(system inflammation response syndrome,SIRS)的... 目的研究低氧诱导大鼠外周血单核细胞(peripheral blood monouclear cells,PBMCs)膜内PKC膜易位及活性变化对肿瘤坏死因子α(TNF-α)表达的作用,探讨缺氧与全身炎症反应综合征(system inflammation response syndrome,SIRS)的关系。方法采用明胶法分离大鼠外周血单核细胞,于低氧条件下(3%O2,5%CO2,92%N2)培养0、1、3、6、9、12、24h后,收集细胞,分别采用免疫沉淀法、SDS-PAGE、PKC活性检测试剂盒及逆转录PCR(RTPCR)检测PKC膜易位、PKC活性及TNF-α表达量,采用SPSS10.0分析以上指标变化的相关性。结果低氧1~9h,大鼠外周血单核细胞膜PKCα含量及活性明显增高(P〈0.01)。低氧1~24h,大鼠外周血单核细胞膜PKCε含量及活性无明显变化。缺氧1~9h,大鼠外周血单核细胞TNF-α mRNA表达明显增高(P〈0.01)。低氧1~24h,PKCα膜蛋白含量及活性变化与TNF-α mRNA表达水平间呈显著正相关(P〈0.01~0.05)。结论低氧可诱导PKCα向膜易位并被激活,活化的PKCα可增强促炎症因子TNF-α的表达。这些变化可能与急性呼吸窘迫综合征(acute respiratory dysfunction,ARDS)时血浆中大量促炎症因子持续存在密切相关。 展开更多
关键词 低氧 pkc 肿瘤坏死因子Α
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基于CRISPR/Cas9技术构建PKCα基因敲除的人腹膜间皮细胞系
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作者 吴忠财 马祖福 +4 位作者 梁望群 左学志 姚颖 徐钢 王艻 《临床肾脏病杂志》 2021年第9期750-754,共5页
目的通过CRISPR/Cas9技术敲除人腹膜间皮细胞系HMrSV5的PKCα基因,为腹膜纤维化的发病机制研究提供体外细胞模型。方法针对人PKCα基因外显子区域设计3对sgRNA序列,其中两对序列靶点位于第4外显子,一对序列靶点位于第3外显子。由引物合... 目的通过CRISPR/Cas9技术敲除人腹膜间皮细胞系HMrSV5的PKCα基因,为腹膜纤维化的发病机制研究提供体外细胞模型。方法针对人PKCα基因外显子区域设计3对sgRNA序列,其中两对序列靶点位于第4外显子,一对序列靶点位于第3外显子。由引物合成公司合成两端含酶切位点黏端的寡聚双链DNA,酶切连接至真核表达载体GV393。使用293T细胞进行慢病毒包装,通过病毒感染HMrSV5细胞,检测HMrSV5细胞中PKCα的敲除效率即脱靶情况。结果3对sgRNA通过慢病毒表达载体成功转染HMrSV5细胞,经实时荧光定量PCR及免疫印迹检测两对sgRNA成功敲低PKCα。错配酶和基因测序结果显示PKCα基因组DNA导入有效突变,3个脱靶位点与原参考序列一致,排除脱靶情况。结论利用CRISPR/Cas9技术成功构建了PKCα敲除的HMrSV5细胞,有利于相关基因体外研究。 展开更多
关键词 腹膜纤维化 pkcΑ CRISPR/Cas9 基因敲除
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Interaction between fragile histamine triad and protein kinase C alpha in human non-small cell lung cancer tissues
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作者 Peng-hui Zhuang1, Zhao-hui Liu2, Xiao-gang Jiang3, Cheng-en Pan41. Department of Thoracic Surgery, the First Affiliated Hospital, Medical School of Xi’an Jiaotong University 2. Department of Anatomy and Histology & Embryology, Medical School of Xi’an Jiaotong University +1 位作者 3. Department of Genetics and Molecular Biology, Medical School of Xi’an Jiaotong University 4. Department of Geratic Surgery, the First Affiliated Hospital, Medical School of Xi’an Jiaotong University, Xi’an 710061, China. 《Journal of Pharmaceutical Analysis》 SCIE CAS 2009年第1期57-61,共5页
Objective To investigate the interaction between fragile histamine triad (FHIT) and protein kinase C alpha (PKCα) in human non-small cell lung cancer tissues. Methods FHIT and PKCα double positive samples were scree... Objective To investigate the interaction between fragile histamine triad (FHIT) and protein kinase C alpha (PKCα) in human non-small cell lung cancer tissues. Methods FHIT and PKCα double positive samples were screened by immunohistochemical staining from 13 human non-small cell lung cancer tissues. Co-immunoprecipitation was performed by using anti-FHIT and anti-PKCα. The immune precipitate was analyzed by SDS-PAGE and Western blot. Results Immune precipitate staining detection showed that 3 samples out of the 13 cases were double positive for FHIT and PKCα. FHIT protein was present in the immune precipitate of anti-PKCα while there was PKCα in the immune precipitate of anti-FHITmAb. Conclusion FHIT and PKCα exist as a complex in human non-small cell lung cancer tissues, which will provide a new route for studying the pathogenesis and immunotherapy of human non-small cell lung cancer. 展开更多
关键词 fragile histamine triad (FHIT) protein kinase C alpha (pkcα) non-small cell lung cancer (NSCLC) CO-IMMUNOPRECIPITATION
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Effect of Combination Regimen of Low-dose Gossypol Acetic Acid with Steroid Hormones on Expression of Protein Kinase C alpha (PKC-α) and Cyclin D1 in Rat Testes 被引量:1
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作者 Jin-hui SHAO Zeng-lu XU +2 位作者 Xiao-jing QIAN Fang LIU Hui HUANG 《Journal of Reproduction and Contraception》 CAS 2012年第4期199-208,共10页
Objective To investigate the contraceptive mechanism of combination regimen of low- dose gossypol acetic acid (GA) with steroid hormones [desogestre/ethinylestradiol/ testosterone undeeanote(DSG/EE/TU)]. Methods A... Objective To investigate the contraceptive mechanism of combination regimen of low- dose gossypol acetic acid (GA) with steroid hormones [desogestre/ethinylestradiol/ testosterone undeeanote(DSG/EE/TU)]. Methods Adult male rats were randomly divided into four groups. Group GH: rats were fed orally with gossypol acetic acid (GA, 12.5 mg/kg) and desogestrel (DSG, 0.125 mg/kg)/ ethinylestradiol (EE, 0.025 mg/kg)/testosterone undecanoate (TU, 100 mg/kg per day, qd X4 weeks or 10 weeks); group G: a single dose of GA (12.5 mg/kg per day, qd X 4 weeks or I0 weeks); group H: the same dosage of DSG/EE/TUas in group GH; group C: rats were treated with vehicle (1% methyl cellulose) as the control. Expression of protein kinase C alpha (PKC-a) and cyclin D1 in rat testes were tested mainly by immunohistochemistry (IHC) and Western blotting. Results IHC results showed that protein PKC- a was expressed mainly in interstitial tissue of testis among seminiferous tubule. The expression of PKC-oc in groups H and GH at week 10 was decreased greatly compared with that in group C. The protein cyclin D1 was expressed mainly in residual body of seminiferous tubule eavosurface and interstitial tissue among seminiferous tubule of testis. Western blotting results showed that the expression of PKC-a in groups H and GH at week 10 was decreased significantly compared with that in group C (P〈0.05). The expression of cyclin D1 in groups G, H or GH at week 10 rose significantly compared with that in group C (P 〈0.05).Conclusion The administration of low-dose gossypol acetic acid with steroid hormones for 10 weeks can decrease the expression of PKC-a greatly. 展开更多
关键词 GOSSYPOL steroid hormones protein kinase C alpha pkc-a) cyclin D 1 RAT TESTES
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