针刺对小鼠腹腔巨噬细胞的热休克蛋白诱导性一氧化氮合酶？…

目的 研究针刺对小鼠腹腔巨噬细胞的热休克蛋白（ＨＳＰ）,诱导性一氧化氮合酶（ｉＮＯＳ）及其ｍＲＮＡ表达的效应。方法 将２４只昆明小鼠腹腔注射无菌石蜡油后,随机分为３组;电针（ＥＡ）组,对照１（Ｃ１）组和对照２（Ｃ２）组。将３组收集的腹腔巨噬细胞制备成玻片和硝酸纤维素（ＮＣＭ）两种标本,应用原位杂交,免疫细胞化学,细胞化学及斑点印变技术检测ＨＳＰ７０,ｉＮＯＳ,及ｉＮＯＳ ｍＲＮＡ。     

针刺对小鼠巨噬细胞基因表达的效应

目的 为了探讨针刺镇痛效应与细胞免疫的关系,研究了针刺对小鼠巨噬细胞中ｃ－ｆｏｓ ｍＲＡＮ ｐｐＥＮＫｍＲＮＡ,ｉＮＯＳｍＲＮＡ及ｉＮＯＳ活性的效应。方法 ２０只ＢＡＬＢ／ｃ小鼠被随机分成２组;ａ．用５Ｈｚ电针处理的针刺组;ｂ．未用电针处理的对照组。针刺或牵拉前后用钾离子渗透法检测痛阈。实验采用原位杂交,ＮＡＤＰＨＮＢＴ组织化学,ＲＮＡ斑点印迹及蛋白质斑点印迹技术。应用ＴＬＣ扫描仪扫描斑点印迹信号并作统计学处理。结果 杂交信号和ｉＮＯＳ活性呈现蓝紫色,信号均定位于巨噬细胞的胞质。与对照组相比,针刺组的所有印迹信号均增强,Ｐ〈０．０１。针刺组中ｃ－ｆｏｓ ｍＲＮＡ,ｐｐＥＮＫｍＲＮＡ,ｉＮＯＳｍＲＮＡ的表达及ｉＮＯＳ的活性与提高的痛阈呈正相关。此外,ｃ－ｆｏｓ ｍＲＮＡ与ｍＲＮＡ与ｐｐＥＮＫｍＲＮＡ之间的变动呈正     

低剂量过氧化氢预处理导致牛肺血管内皮细胞耐受过氧化氢损伤

小量过氧化氢（Ｈ２Ｏ２）预处理，能诱导牛肺动脉内皮细胞（ＢＰＡＥＣｓ）中热休克蛋白７０（ＨＳＰ７０）及ＨＳＰ７０ｍＲＮＡ增多，同时使ＢＰＡＥＣｓ获得了对大量Ｈ２Ｏ２损伤的耐受性，表现为大量Ｈ２Ｏ２所致乳酸脱氢酶释放和硫代巴比妥酸反应物含量增加及过氧化氢酶、超氧化歧化酶活性降低等变化减轻。用放线菌酮和放线菌素Ｄ分别抑制ＨＳＰ７０和ＨＳＰ７０ｍＲＮＡ的增多后，此种耐受性消失，说明小量Ｈ２Ｏ２预处理使Ｂ     

缺氧对肺动脉内皮细胞和平滑肌细胞5—羟色胺转载体基因表达 …

目的：探讨无氧（０％Ｏ２＋９５％Ｎ２＋５％ＣＯ２）和低氧（２．５￣３％＋９％Ｎ２＋５％ＣＯ２）对新生小牛肺动脉内皮细胞（ＰＡＥＣ）和肺动脉平滑肌细胞（ＰＡＳＭ）５－羟色胺转载体基因表达的影响。方法：应用细胞培养，核到分子杂交技术。结果：无氧组和低氧组（１．５ｈ、３ｈ、６ｈ）ＰＡＥＣ５－羟色胺转载体ｍＲＮＡ表达显著同于常氧组（Ｐ〈０．０５），而无氧和低氧１２ｈ至４８ｈ对ＰＡＥＣ５－羟色胺转载体ｍＲＮ     

热休克基因表达对抗过氧化氢所致肺内皮细胞损伤的保护作用

本研究探讨热休克反应（ｈｅａｔｓｈｏｃｋｒｅｓｐｏｎｓｅ，ＨｓＲ）对抗过氧化氢（Ｈ２Ｏ２）所致牛肺动脉内皮细胞（ＢＰＡＥＣｓ）损伤的保护作用及机制。实验发现，预先热休克处理（４２℃，２ｈ）可使ＢＰＡＥＣｓ中热休克蛋白７０ｋＤａ（ＨＳＰ７０）及ＨＳＰ７０ｍＲＮＡ明显增多，同时显著减轻Ｈ２Ｏ２所致的ＢＰＡＥＣｓ中乳酸脱氢酶释放和硫代巴比妥酸反应物含量增加及过氧化氢酶和超氧化物歧化酶活性的降低等变化。进一步实验证明放线菌酮和放线菌素Ｄ能分别抑制热休克诱导的Ｈ８Ｐ７０和ＨＳＰ７０ｍＲＮＡ的增多，同时均能取消ＨＳＲ对抗Ｈ２Ｏ２所致ＢＰＡＥＣｓ损伤的保护作用。结果提示，ＨＳＲ具有对抗Ｈ２Ｏ２所致ＢＰＡＥＣｓ损伤的保护作用；此种保护作用与细胞经热休克处理后细胞中热休克基因在转录和翻译两个水平的表达增强及抗氧化能力增加有关。     

一氧化碳对大鼠血管平滑肌细胞低氧性增殖反应的作用

目的：观察低氧时大鼠肺动脉平滑肌细胞（ＰＡＳＭＣ）受血管舒张因子一氧化碳的作用。方法：随机分为常氧组、低氧组（１％Ｏ２＋５％ＣＯ２＋９４％Ｎ２）、一氧化碳组（３％ＣＯ＋５％ＣＯ２＋９２％Ｎ２）,采用流式细胞仪、增殖细胞抗原（ＰＣＮＡ）、「６３Ｈ」－胸腺嘧啶「＾３Ｈ」－ＴｄＲ摄取量,检测ＰＡＳＭＣ增生的情况。结果：（１）流式细胞仪检测低氧组ＰＡＳＭＣＣ２／Ｍ期细胞比例明显增多,Ｇ０／Ｇ１期细胞比例明     

慢性缺氧影响肺脏内皮素和一氧化氮合成酶mRNA的表达

以逆转录多聚酶链反应（ＲＴ－ＰＣＲ）检测了慢性缺氧大鼠肺组织内内皮素－１（ＥＴ－１）ｍＲＮＡ．内皮素受体－Ａ（ＥＴＲ－Ａ）ｍＲＮＡ和一氧化氮合成酶（ＮＯＳ）ｍＲＮＡ的表达水平。结果显示：缺氧１，２．３周时，肺内ＥＴ－１ｍＲＮＡ分别较正常对照上升６３．８％，１０５．５％和４０．９％（Ｐ均＜０．０５）。ＥＴＲ－ＡｍＲＮＡ分别上升５０．６％．５７．４％和５６．３％（Ｐ均＜０．０５），而ＮＯＳｍＮＲＡＲ则分别较正常对照下降３４．２％．４９．６％和３３．５％（Ｐ均＜０．０５）。提示缺氧时肺内ＥＴ水平上升和ＮＯ减少与缺氧刺激肺内ＥＴ－１ｍＲＮＡ表达增加和ＮＯＳｍＲＮＡ表达减少有关。     

慢性缺氧影响肺脏内皮素和一氧经氮合成酶mRNA的表达

以逆转录多聚酶链反庆（ＲＴ－ＰＣＲ）检测了慢性缺氧大鼠肺组织内皮素－１（ＥＴ－１）ｍＲＮＡ内皮素受体－Ａ（ＥＴＲ－Ａ）ｍＲＮＡ和一氧化氮合成酶（ＮＯＳ）ｍＲＮＡ的表达水平，结果显示：缺氧１、２、３周时，肺内ＥＴ－１ｍＲＮＡ分别较正常对照上升６３．８％，１０５．５％，和４０．９％（Ｐ均〈０．０５），ＥＴＲ－ＡｍＲＮＡ分别上升５０．６％，５７．４％和５６．３％（Ｐ均〈０．０５）而ＮＯＳｍＲＮＡ则分别较     

香烟烟雾提取物对大鼠肺组织的损伤作用及对一氧化氮生成的影响

将大鼠肺组织切片与香烟烟雾提取物（ＣＳＥ）共同孵育,测定其乳酸脱氢酶（ＬＤＨ）活性、亚硝酸盐（ＮＯ_２） 含量、一氧化氮合酶（ＮＯＳ）活性及左旋精氨酸（Ｌ－Ａｒｇ）转运,以探讨吸烟对肺组织的氧化损伤作用及对一氧化氮 生成的影响。结果显示： １０％ ＣＳＥ时间依赖地增加大鼠肺组织 ＬＤＨ漏出及 ＮＯ_２释放,二者高度正相关（ｒ=０．６５, Ｐ＜０．０１）,刺激诱导型ＮＯＳ（ｉＮＯＳ）活性增加,并通过增加最大转运速度（Ｖｍａｘ）使Ｌ－Ａｒｇ转运增加。提示：吸烟可 能通过增加 Ｌ－Ａｒｇ的跨膜转运使细胞内 Ｌ－Ａｒｇ浓度增高,并增加 ｉＮＯＳ活性使 ＮＯ过量产生。 ＮＯ可能参与了吸烟对肺 组织的损伤过程。     

病毒激活巨噬细胞产生的一氧化氮与细胞毒的关系

目的：确定新城鸡瘟病毒Ｌｏｄｏｔａ系（ＮＤＶ－Ｌ）能否诱导小鼠腹腔巨细胞（ＰＥＭψ）产生一氧化氮（ＮＯ）以及该 小鼠ＰＥＭψ介导的细胞毒作用是否有依赖关系。方法：采用Ｇｒｉｅｓｓ、ＦＡＣＳ分析和噻唑蓝（ＭＴＴ）方法。结果：ＮＣＶ－Ｌ与ＰＥＭψ任用２Ｈ后就可以稳定地吸附在ＰＥＭψ表面,吸附率达８０％左右;收集ＮＣＶ－Ｌ与ＰＥＭψ作用２４Ｈ上清,发现有ＮＯ的释,释放量与阳性对照组（ＢＸＣＧ－ＬＰＳ－Ｐ     

Dietary selenium supplementation alleviates immune toxicity in the hearts of chickens with lead-added drinking water

Lead (Pb) is an environmental pollutant and can damage organisms. Selenium (Se) can alleviate Pb poisoning. The present study aimed to investigate the alleviative effect of Se on Pb-induced immune toxicity in chicken hearts. One-hundred-and-eighty Hy-line male chickens were randomly divided into four groups at 7 days of age. The control group was offered a standard commercial diet (SD) and drinking water (DW); the Se group was offered SD supplemented with sodium selenite (SeSD) and DW; the Pb?+?Se group was offered SeSD and DW supplemented with lead acetate (PbDW); and the Pb group was offered SD and PbDW. Relative mRNA expression of inducible nitric oxide synthase (iNOS), interleukins (IL-2, IL-4, IL-6, IL-12β, IL-17 and IFN-γ), and heat shock proteins (HSP27, HSP40, HSP60, HSP70, and HSP90) were determined by means of quantitative real-time PCR. Relative protein expression of iNOS, HSP60, HSP70, and HSP90 was assessed, as well as nitric oxide (NO) content and iNOS activity in heart tissue. The results indicated a down-regulation of interleukin (IL)-2 and IFN-γ and an up-regulation of NO, iNOS, interleukins (IL-4, IL-6, IL-12β, IL-17), and heat shock proteins (HSP27, HSP40, HSP60, HSP70, and HSP90) in Pb-damaged hearts. Se alleviated all of the above Pb-induced changes. There were time-dependent effects on NO content, iNOS activity, and mRNA levels of iNOS, IL-2, IL-4, IL-6, IL-17, HSP27, HSP40, HSP60, HSP70, and HSP90 after Pb treatment in the chicken hearts. Se alleviated Pb-induced immune toxicity in the chicken hearts.     

8-Br-cAMP对人视网膜母细胞瘤HXO-Rb44细胞抑癌基因表达的效应

目的 探讨８－Ｂr－CＡＭＰ对人视网膜母细胞瘤之ＨＸＯ－Ｒb４４细胞抑癌基因表达的 效庆及其对细胞生长的影响。方法 应用原位杂交、ＲＮＡ斑点印迹检测Ｐ１６＾ＩＮＫ４mＲＮＡ、p２１＾ＷＡＦ１mＲＮＡ、wp５３mＲＮＡ、mp５３mＲＮＡ和ＲｂｍＲＮＡ,应用免疫组织和化学和蛋白质斑点印迹技术检测ＰＣＮＡ－ＩＲ、p２１＾ＷＡＦ１－ＩＲ、 ｐ１６－ＩＲ、ｐＲｂ－ＩＲ、ｃｄｋ２、ＩＲ和ｃｄｋ４－ＩＲ。结果 在人ＨＸＯ－Ｒ     

半枝莲黄酮对Aβ 25-35引起的大鼠皮层星形胶质细胞NOS、HSP70及apoE异常表达的影响

 目的:探讨半枝莲黄酮对Aβ 25-35引起大鼠皮层星形胶质细胞一氧化氮合酶（NOS）热休克蛋白70（HSP70）及载脂蛋白E（apoE）蛋白表达异常变化的影响。方法:培养第3代的大鼠星形胶质细胞随机分为空白对照组、模型组和3个剂量药物组,药物组细胞分别加入17.5、35和70 mg/L半枝莲黄酮作用24 h后,模型组和3个剂量药物组均加入Aβ 25-35 100 μmol/L作用24 h,免疫组化法测定星形胶质细胞中内皮型一氧化氮合酶 (eNOS)、诱导型一氧化氮合酶 (iNOS) 和神经元型一氧化氮合酶 (nNOS)蛋白的表达;Western blotting检测星形胶质细胞中HSP70蛋白的表达; RT-PCR测定细胞中apoE mRNA的表达。结果:与空白组相比,模型组eNOS蛋白含量降低60.83%（P<0.01）,iNOS蛋白含量增加215.03%（P<0.01）,HSP70蛋白含量增加166.67%（P<001）,apoE mRNA含量增加150.00% (P<0.01);半枝莲黄酮17.5、35及70 mg/L能不同程度地提高eNOS蛋白含量73.66%～137.77% (P<0.05),降低iNOS蛋白含量19.40%～44.50%（P<0.01）,降低HSP70蛋白含量1852%～49.38%（P<0.01）,降低apoE mRNA的含量14.17%～41.67% (P<0.01)。结论:半枝莲黄酮对Aβ 25-35引起的大鼠皮层星形胶质细胞的损伤具有抑制作用。半枝莲黄酮可能通过影响星形胶质细胞发挥对阿尔茨海默病的治疗作用。     

Distribution of 70kDa heat shock protein in rabbit brains after heat stress and heat stroke

OBJECTIVE: Evaluation of the relationship between the induction of 70kDa heat shock protein in rabbit brains and heat stress. METHODS: HSP70 was detected using monoclonal antibody by ABC method in rabbit hypothalamus, hippocampus and cerberal cortex. RESULTS: Intense HSP70 staining was displayed in rabbit brains of the heat stroke group (rectal temperature 43 degrees C to death). Positive cells were distributed mainly in the CA1, CA2 regions of the hippocampus; granular cell layer I and pyramidal layer (II) of the cerebral cortex; and the periventricular area of hypothalamus. HSP70-psoitive substances were localized in the cytoplasm and neuronal processes, a few neurons exhibited dark staining nucle. Hosever, the rabbit brains of the general heat stress group (rectal temperature 42.0 degrees C, 30 minutes) had much weaker staining. CONCLUSION: Hyperthermia causes neuronal expression of HSP70, particularly under strong heat stress, and may be sustained till death.     

Expression of inducible nitric oxide synthase in rat pulmonary Cryptococcus neoformans granulomas.

Rats, like humans, have extremely effective immune mechanisms for controlling pulmonary Cryptococcus neoformans infection. The mechanism(s) responsible for efficient immunity in rat experimental infection is unknown. Recently, induction of inducible nitric oxide synthase (iNOS) and nitric oxide (NO) have been implicated as an important microbicidal mechanism by which activated macrophages effect cytotoxicity against microbes. In this report, we investigated the expression of iNOS in rat pulmonary cryptococcosis. Localization and regulation of NO production was studied by immunohistochemistry for iNOS in conjunction with immunohistochemistry for cell markers, cytokines, and cryptococcal capsular polysaccharide. iNOS immunoreactivity was detected in macrophages, neutrophils, vascular endothelium, and respiratory epithelium. Double-immunolabeling studies revealed that the most prominent iNOS immunoreactivity was localized to epithelioid macrophages (CD11b/c+) within granulomas; CD4+ and CD8+ T cells were numerous around granulomas but did not express iNOS. iNOS immunoreactivity was detected in a selective population of epithelioid macrophages within some granulomas but not others. iNOS- granulomas were identical to iNOS+ granulomas with respect to morphology and immunohistochemical profiles. Macrophage iNOS immunoreactivity was detected 1 week after infection in one out of four rats and was strongly expressed in all rats at 2 weeks (in up to 50 percent of the granulomas) but declined considerably by 25 days. iNOS expression coincided with granuloma formation and preceded a decrease in lung fungal burden, suggesting an anticryptococcal role for NO. By double labeling, cytokines that have been shown to promote (interferon-gamma, granulocyte/macrophage colony-stimulating factor) and inhibit (transforming growth factor-beta) macrophage iNOS expression were detected around iNOS+ granuloma. iNOS immunoreactivity was expressed in selected neutrophils (1 and 2 weeks) and endothelial cells (1 and 2 weeks and 25 days) in the inflamed lung. Airway iNOS immunoreactivity was limited to the luminal border of rare bronchiolar epithelial cells. iNOS immunoreactivity was not detected in uninfected rats. The present study provides the first evidence for association of iNOS expression with protective cellular responses to cryptococcal infection in vivo.     

Myocardial tolerance to ischemia-reperfusion injury, training intensity and cessation

Training has been shown to induce cardioprotection. The mechanisms involved remain still poorly understood. Aims of the study were to examine the relevance of training intensity on myocardial protection against ischemia/reperfusion (I/R) injury, and to which extent the beneficial effects persist after training cessation in rats. Sprague-Dawley rats trained at either low (60% [(V)\dot]\textO_2max {\dot{{V}}\text{O}}_{2\max } ) or high (80% [(V)\dot]\textO_2max {\dot{{V}}\text{O}}_{2\max } ) intensity for 10 weeks. An additional group of highly trained rats was detrained for 4 weeks. Untrained rats served as controls. At the end of treatment, rats of all groups were split into two subgroups. In the former, rats underwent left anterior descending artery (LAD) ligature for 30 min, followed by 90-min reperfusion, with subsequent measurement of the infarct size. In the latter, biopsies were taken to measure heat-shock proteins (HSP) 70/72, vascular endothelial growth factor (VEGF) protein levels, and superoxide dismutase (SOD) activity. Training reduced infarct size proportionally to training intensity. With detraining, infarct size increased compared to highly trained rats, maintaining some cardioprotection with respect to controls. Cardioprotection was proportional to training intensity and related to HSP70/72 upregulation and Mn-SOD activity. The relationship with Mn-SOD was lost with detraining. VEGF protein expression was not affected by either training or detraining. Stress proteins and antioxidant defenses might be involved in the beneficial effects of long-term training as a function of training intensity, while HSP70 may be one of the factors accounting for the partial persistence of myocardial protection against I/R injury in detrained rats.     

双歧杆菌的完整肽聚糖对LPS激活裸鼠腹腔巨噬细胞的影响

目的 了解被ＬＰＳ激活的裸鼠腹腔巨噬细胞在用双歧双歧杆菌的完整聚糖刺激后产生的ＮＯ、ｉＮＯＳ及ｃＧＭＰ的水平。方法 以Ｇｒｉｅｓｓ试剂、激光截聚焦显微镜以及入免法分别测定裸鼠腹腔巨噬细胞产生的ＮＯ、ｉＮＯＳ及ｃＧＭＰ的水平。结果 完整肽聚糖注射组裸鼠腹腔巨噬细胞在ＬＰＳ诱导下产生ＮＯ、ｉＮＯＳ及ｃＧＭＰ含量均显著高于对照组（Ｐ〈０．０１）。结论 双歧双歧杆菌的完整肽聚糖能协同ＬＰＳ激活巨噬细胞,使     

急性心肌梗死猝死者心肌细胞hsf1和HSP70改变的意义

目的探讨急性心肌梗死猝死者梗死区心肌细胞内热休克转录因子1（hsfl）和热休克蛋白70（HSP70）改变的临床意义。方法分别用RT-PCR和免疫组化法检测（IHC）18例急性心肌梗死猝死者（研究组）和15例心脏正常因车祸快速死亡者（对照组）心肌细胞中hsfl和HSP70基因的mRNA和蛋白表达量。结果急性心肌梗死猝死者心肌细胞hsfl和HSP70的mRNA表达量都显著高于正常对照组（P〈0.01），且hsfl和HSP70mRNA的表达量之间呈显著的正相关关系（P〈0．001）。急性心肌梗死猝死者心肌细胞hsfl和HSP70蛋白在细胞浆和细胞核表达较对照组显著增强（P〈0.001），其中hsfl蛋白主要在心肌细胞核内表达，HSP70蛋白主要在心肌细胞浆内表达。结论急性心肌梗死猝死者心肌细胞内hsfl和HSPT0可能共同参与了急性心肌梗死的病理生理过程．这一过程可能是热休克反应的另一调节途径。