连翘酯苷对顺铂作用后豚鼠耳蜗c-jun表达的影响

目的：研究连翘酯苷对顺铂作用后豚鼠耳蜗c—jun表达的影响。方法：将30只豚鼠随机分为对照组（10只）,顺铂组（10只）和连翘酯苷组（10只）。腹腔注射顺铂溶液（8mg／kg）,1次／d,连续7d,建立顺铂耳毒性模型;连翘酯苷组在每次注射顺铂溶液30rnin前腹腔注射连翘酯苷25．0mg／kg／d,连续7d;对照组以生理盐水代替顺铂溶液注射,连续7d。实验动物被处死前,检测其DPOAE幅值变化;采用蛋白质印迹杂交（Western Blotting）检测各组豚鼠耳蜗c—jun蛋白的表达,逆转录聚合酶链反应检测各组豚鼠耳蜗c—jun基因mRNA的表达。结果：顺铂组DPOAE幅值明显低于对照组（P〈0．01）;相比于顺铂组,连翘酯苷组DPOAE幅值明显升高（P〈0．05）。顺铂组豚鼠耳蜗c—jun蛋白与mRNA表达水平均显著高于对照组（P〈0．01）;相比于顺铂组,连翘酯苷组C-jun蛋白与mRNA表达水平明显降低（P〈0．05）。结论：连翘酯苷能够通过降低c—jun的表达防护顺铂所致的耳蜗损伤。     

生长因子对HSC-T6细胞c-fos和c-jun基因表达的影响

目的 了解血小板衍生生长因子(PDGF)和转化生长因子β(TGF β)分别对肝星状细胞c～fos和c-jun基因表达的影响.方法 在培养的肝星状细胞系HSC-T16细胞中分别加入不同浓度的PDGF(终浓度分别为8、40、200 ng/ml)和TGF β(终浓度分别为0.2,1.0、5.0 ng/ml);于8、24,48、72h四个时间点分别收集细胞,提取细胞总RNA;用逆转录定量PCR法测定c～los和c-jun的基因表达水平.结果 PDGF处理的3组HSC-T6细胞8、24、48、72h时,c-fos基因表达水平均明显高于对照组,并呈剂量依赖性l培养8 h时达到表达最高峰,对照组,PDGF 8 ng/ml组,PDGF 40 ng/ml组,PDGF 200ng/ml组c-fos表达分别为0.63±0.13,1.13±0.19、1.75±0.20、2.40±0.23,3组间差异有统计学意义(F=7.03,P<0.01).TGF β处理的3组HSC-T6细胞8、24,48、72 h时,c-iun基因表达水平均明显高于对照组,并呈剂量依赖性,培养8 h时达到表达最高峰,对照组,TGF β 0.2ng/ml组、TGF β 1.0ng/ml组、TGF β 5.0ng/ml组cjun基因表达分别为0.93±0.13、1.69±0.26、2.34±0.30、2.96士0.37; 3组间差异有统计学意义(F=6.34,P<0.01).结论 PDGF和TGF β分别对肝星状细胞c-los和c-jun基因表达有明显的上调作用.     

地尔硫卓对增殖血管平滑肌细胞的原癌基因表达的影响

目的观察地尔硫卓（Dil）对增殖血管平滑肌细胞（VSMC）的原癌基因c-myc、c-fos、c-jun和ras mRNA表达的影响。方法将组织贴块法培养的大鼠胸主动脉VSMC随机分为5组,即空白组、模型组和Dil1、2、3组（浓度分别为10^-5、10^-6、10^-7mol/L）,应用MTT检测增殖能力,用流式细胞术检测VSMC的增殖指数,用rt-PCR检测c-myc、c-fos、c-jun和ras mRNA的表达。结果与模型组比较各浓度Dil都能抑制VSMC增殖,PI值均显著下降（各组PI值分别为21.53±1.72、28.63±0.96、15.95±0.37、19.28±0.94、20.33±0.67;P〈0.05）;Dil1、2组c-myc、c-fos、c-jun mRNA的表达显著减少（模型组和Dil1、2组的c-myc/GAPDH值分别为2.454±0.03、1.509±0.05、1.660±0.04,c-fos/GAPDH值分别为0.0046±0.0004、0.0023±0.0003、0.0038±0.0005,c-jun/GAPDH值分别为1.950±0.03、1.077±0.03、1.725±0.03;P〈0.05）,rasmRNA的表达显著增加（模型组和Dil1、2组的ras/GAPDH值分别为1.941±0.03、3.811±0.02、2.501±0.02;P〈0.05）。结论Dil抑制VSMC增殖机制可能与下调原癌基因c-myc、c-fos、c-jun的表达和上调ras的表达有关。     

An association between ethanol-evoked enhancement of c-jun gene expression in the nucleus tractus solitarius and the attenuation of baroreflexes

BACKGROUND: An increased expression of Fos, the protein product of the immediate early gene c-fos, in the nucleus tractus solitarius (NTS) is associated with dysfunction in baroreceptor reflex control of heart rate. Our previous studies demonstrated that ethanol attenuates baroreflex sensitivity (BRS) in rats and humans. In this study, we tested the hypothesis that enhanced expression of the immediate early gene c-jun (an index of neuronal activity) in the NTS contributes to the baroreflex dysfunction caused by ethanol. METHODS: Conscious male spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) rats were used to measure blood pressure, heart rate, and baroreflex sensitivity (Oxford method). The c-jun messenger RNA (mRNA) expression in NTS was measured by in situ hybridization. RESULTS: Ethanol elicited dose-dependent attenuation in BRS in WKY rats, which was associated with significant increases in c-jun mRNA in the NTS. In contrast, ethanol had no effect on BRS or c-jun mRNA in the NTS of the SHRs; the latter exhibited significantly lower BRS and higher c-jun mRNA in the NTS compared with WKY rats. CONCLUSIONS: An increased basal level of c-jun mRNA in the NTS may contribute to the reduced BRS in the SHR, and ethanol enhancement of neuronal activity of the NTS, expressed as increased c-jun mRNA expression, may contribute to its attenuation of BRS, which highlights the NTS as a neuroanatomical target for ethanol action on baroreflexes.     

The hinge region in androgen receptor control

The region between the DNA-binding domain and the ligand-binding domain of nuclear receptors is termed the hinge region. Although this flexible linker is poorly conserved, diverse functions have been ascribed to it. For the androgen receptor (AR), the hinge region and in particular the (629)RKLKKL(634) motif, plays a central role in controlling AR activity, not only because it acts as the main part of the nuclear translocation signal, but also because it regulates the transactivation potential and intranuclear mobility of the receptor. It is also a target site for acetylation, ubiquitylation and methylation. The interplay between these different modifications as well as the phosphorylation at serine 650 will be discussed here. The hinge also has an important function in AR binding to classical versus selective androgen response elements. In addition, the number of coactivators/corepressors that might act via interaction with the hinge region is still growing. The importance of the hinge region is further illustrated by the different somatic mutations described in patients with androgen insensitivity syndrome and prostate cancer. In conclusion, the hinge region serves as an integrator for signals coming from different pathways that provide feedback to the control of AR activity.     

c-jun siRNA的构建及其对胃癌细胞株SGC7901中β3GnT8基因表达的影响

目的 构建c-jun干扰载体,有效沉默胃癌SGC7901细胞株中的c-jun基因表达,研究其对胃癌SGC7901细胞株中β3GnT8基因表达的影响.方法 ①利用互联网资源针对c-jun基因的靶序列设计合成四条siRNA,将四条siRNA插入到RNA干扰载体pGPU6/GFP/Neo中构建成四条干扰载体.②将构建好的c-jun siRNA载体质粒pGPU6/GFP/Neo-sic-jun-1949、pGPU6/GFP/Neo-sic-jun-1050、pGPU6/GFP/Neo-sic-jun-1111、pGPU6/GFP/Neo-sic-jun-1276通过阳离子脂质体将表达质粒分别转染SGC7901细胞,48h后荧光显微镜下观察不同浓度质粒的转染效率,通过RT-PCR法检测c-jun和β3GnT8基因mRNA的表达.结果 ①成功构建了四条正确的c-jun siRNA表达质粒;②成功筛选出一条对c-jun有明显抑制作用的siRNA干扰质粒,阻断c-jun表达后,糖基转移酶β3GnT8的表达受到明显抑制.结论 构建的c-jun siRNA表达质粒可以有效抑制胃癌SGC7901细胞株的c-jun基因表达,同时抑制糖基转移酶β3GnT8的表达,为研究胃癌的分子机制提供了一定的理论基础.     

疏肝调神针法对偏头痛大鼠脑中c-fos、c-jun表达的影响

目的:通过颈后皮下注射硝酸甘油构建偏头痛模型大鼠,观察疏肝调神针法对偏头痛模型大鼠中脑c-fos、c-jun基因、蛋白的表达,探讨疏肝调神针法对偏头痛防治的作用机制。方法:Wistar大鼠按照随机数字表法分为对照组、模型组、普通针刺组、疏肝调神组。采用颈后皮下注射硝酸甘油制备偏头痛模型。疏肝调神组取百会、风池(双)、内关(双)、太冲(双)、普通针刺组取百会、风池(双)进行针刺,余组只抓取固定。分别采用荧光定量PCR法、Western Blot法检测大鼠中脑c-fos、c-jun基因、蛋白的表达。结果:与对照组相比,模型组大鼠中脑c-fos、c-jun基因、蛋白表达均显著升高(P <0. 01);与模型组比较,疏肝调神组、普通针刺组大鼠中脑c-fos、c-jun基因、蛋白表达均有明显降低(P <0. 01);与普通针刺组比较,疏肝调神组大鼠中脑c-fos、c-jun基因、蛋白表达均有不同程度降低(P <0. 01)或(P<0. 05)。结论:针刺可以下调偏头痛模型大鼠中脑部c-fos、c-jun基因、蛋白的表达,抑制三叉神经血管系统痛觉信号的传导,缓解偏头痛的发生,而疏肝调神针法在抑制c-fos、c-jun基因、蛋白的表达上更具优势。     

Effect of brain-derived neurotrophic factor on c-jun expression in the rd mouse retina

AIM: To determine the location of c-jun protein, dynamic changes in c-jun mRNA and protein expression, and ultrastructure characteristics in the rd mouse retina, following a single dose of brain-derived neurotrophic factor (BDNF) in a short period of time. METHODS: A single intravitreal injection of BDNF at two dosages (25μg/L or 50μg/L) was given to the right eye of the rd mouse at age 2 and 3 weeks respectively. Two weeks after injection, the location of c-jun protein in the retina was observed by immunofluorescence detection, c-jun mRNA and protein expression in retinas were detected by quantitative real time polymerase chain reaction (RT-PCR) and western immunoblotting analysis, ultrastructure characteristics of retinas were detected by transmission electron microscope (TEM) observation. RESULTS: c-jun protein was expressed in the inner nuclear layer (INL) of retina. BDNF at two dosages (25μg/L and 50μg/L) increased c-jun mRNA expression at PN-4 weeks respectively (P1=0.019, P2= 0.021). 50μg/L BDNF increased c-jun protein expression at PN-4 weeks (P =0.000). The retinal ultrastructure was improved. CONCLUSION: The effects of BDNF exerts on the c-jun expression in the retina are dose-dependent and time-dependent, which may mediate photoreceptor rescue indirectly in the pathological process of retinitis pigmentosa (RP) at early stage.     

氧化应激在砷中毒中作用研究进展

  目的  探讨亚急性砷中毒对大鼠脑神经氧化应激影响及机制。  方法  清洁级SD大鼠56只,随机分为4组,即对照组（蒸馏水）及低、中、高剂量染砷组（2、10、50 mg/L亚砷酸钠溶液）,雌雄各半,每组14只,以自由饮水方式染砷30 d。用电感耦合等离子体发射光谱（ICP-AES）测定大鼠血砷、脑砷浓度,用5,5 – 二硫二硝基苯甲酸速率微板法测定脑组织还原型谷胱甘肽（GSH）含量,用微量定磷法测定脑组织中γ – 谷氨酰半胱氨酸合成酶（γ-GCS）活性,免疫组化法测定脑海马和皮质区c-fos和c-jun蛋白表达。  结果  与对照组比较,各剂量染砷组大鼠血砷、脑砷浓度明显升高,差异均有统计学意义（P < 0.05）;脑海马区神经元肿胀率、变性率、坏死率高于对照组;与对照组比较,低、中、高剂量染砷组大鼠脑组织中γ-GCS活性[分别为(4.79 ± 0.60)、(3.71 ± 0.87)、(3.13 ± 0.43) U/mgpro]、GSH含量[分别为（29.59 ± 1.73）、（24.57 ± 1.53）、（20.59 ± 2.51）μmol/gpro]明显下降,差异均有统计学意义（均P < 0.05）;与对照组比较,低、中、高剂量染砷组大鼠海马和皮质c-fos阳性率[分别为（65.36 ± 6.38）%、（71.51 ± 6.05） %、（72.20 ± 6.13）%和（67.81 ± 7.47） %、(70.99 ± 6.67) %、(70.77 ± 4.79) %],c-jun阳性率[分别为（56.67 ± 8.29）%、（65.65 ± 5.70）%、（69.88 ± 2.85）%和（59.46 ± 6.05）%、（65.80 ± 4.31）%、（67.51 ± 3.84）%]明显升高,差异均有统计学意义（均P < 0.05）。  结论  亚急性砷中毒导致脑组织c-fos和c-jun蛋白表达增加、γ-GCS活性下降、GSH含量减少、降低脑组织氧化应激能力,可能是亚急性砷暴露致中枢神经毒性作用机制之一。