外源性皮质酮对大鼠主动脉内皮细胞热休克蛋白90表达的影响

目的在细胞水平观察皮质酮对热休克蛋白90（HSP90）表达的影响及量效动态变化。方法采用培养的大鼠主动脉内皮细胞,给予不同干预剂量的皮质酮（0.1μM、50μM和100μM）干预,在2～72h的不同时段检测HSP90的表达水平。结果皮质酮干预大鼠主动脉内皮细胞,HSP90蛋白表达在6h达到高峰,其中干预浓度为0.1μM组HSP90的蛋白含量增加最为明显,到72h时与正常对照组已无显著差异。结论不同干预剂量的皮质酮（0.1μM、50μM和100μM）均可促使大鼠主动脉内皮HSP90表达增加,应激时大量分泌的皮质酮可能是大鼠主动脉内HSP90蛋白表达增加的主要始动环节之一。     

汉滩病毒体外诱导热休克蛋白70的表达

目的定量研究汉滩病毒（HTNV）体外诱导细胞热休克蛋白70（HSP70）及HSP70 mRNA的表达规律。方法以HTNV敏感细胞株Vero-E6为对象，体外实验性感染Hantaan 76—118。采用原位杂交、免疫细胞化学染色法及逆转录聚合酶链反应（RT-PCR）观察感染后72h内HSP70的表达并加以定量分析。结果与模拟感染组相比，Hantaan 76—118感染Vero-E6细胞后0．5h，HSP70 mRNA开始升高，于感染后12h达高峰，并持续高表达至72h（P＜0．05）。Hantaan 76—118感染后2h可检测到HSP70蛋白表达增加，于感染后12h达高峰，并持续高表达至72h（P＜0．05）。结论体外感染HTNV可诱导Vero-E6细胞产生热休克反应并表达HSP70，这种快速、持久的应激反应，可能对HSP70持续发挥其细胞保护作用具有重要意义。     

半夏泻心汤加减方对胃溃疡大鼠胃组织热休克蛋白27表达的影响

[目的]探讨半夏泻心汤加减方调控热休克蛋白27（HSP27）干预乙酸性胃溃疡（Gu）大鼠的疗效机制。[方法]30只Wista大鼠,随机分为假手术对照（S）组、GU模型对照（M）组及半夏泻心汤加减方干预（Y）组,每组10只。M及Y组采用胃窦部乙酸注射复制大鼠乙酸性GU模型,s组注射同剂量的0．85％氯化钠。于术后第2天Y组经灌胃给予半夏泻心汤加减方每日1次,共7d,S、M组给予等量0．85％氯化钠,于术后第8天处死全部大鼠取材,观察胃大体情况,采用GU指数（GUD对溃疡进行评定,苏木精一伊红染色观察胃组织病理变化,实时荧光定量PCR法检测大鼠胃窦部HSP27mRNA表达。[结果]S组大鼠胃腔无溃疡形成,M及Y组大鼠胃腔乙酸注射部位均见溃疡形成,但Y组GUI显著低于M组（P〈0．01）;Y组胃组织HSP27mRNA较S、M组表达显著升高（均P〈0．01）[结论]半夏泻心汤加减方可上调HSP27mRNA表达,这可能是其有效干预乙酸性GU大鼠的病理机制。     

大剂量甲基强的松龙对大鼠脑I／R损伤的保护作用及其机制探讨

目的观察大剂量甲基强的松龙（MP）对大鼠脑缺血／再灌注（I／R）损伤的保护作用,并探讨其机制。方法将48只SD大鼠随机分为A、B组各24只,均制备大鼠局灶性脑I／R模型,A、B组大鼠经尾静脉分别一次性注入MP10μl（90mg／kg）、生理盐水10μl。采用开阔法观察大鼠行为学变化,采用TUNEL法和免疫组织化学方法检测大鼠脑缺血区神经细胞的凋亡和热休克蛋白（HSP27）、细胞色素C-3（Cyt-3）、Caspase-9蛋白。结果与B组相比,MP组SD大鼠探索活动显著增加,脑缺血区凋亡细胞数明显减少,HSP27表达增加,Cyt．3、Caspase-9蛋白表达降低,P均〈0．01。结论大剂量甲基强的松龙对脑I／R损伤有保护作用,可能与其调控HSP27／DISC信号复合体的表达有关。     

急性心肌缺血再灌注对大鼠脑组织损伤及发生机制的探讨

结扎大鼠左冠状动脉前降支建立急性心肌缺血再灌注模型,采用免疫组织化学方法观察大脑、小脑、脑干延髓部、海马等部位热休克蛋白70（HSP70）表达的改变.结果心肌缺血再灌注0.5h后即可在大鼠大脑、小脑、海马和延髓等部位的神经细胞和血管内皮细胞见阳性HSP70表达,随着再灌注时间延长表达增强.心肌缺血再灌注后不同时间段的血压明显降低,脑血流速度明显下降,以心肌缺血后和再灌注30min时下降较为明显.     

糖尿病大鼠肾脏热休克蛋白70的表达及普罗布考干预的研究

目的 观察糖尿病大鼠肾脏中热休克蛋白70（HSP70）的表达并观察普罗布考对HSP70表达的影响. 方法 链脲佐菌素构建糖尿病大鼠模型50只,随机分为糖尿病（DM）组、普罗布考（PT）组,另设健康对照（NC）组.RT-PCR测定各组肾组织HSP70 mRNA的表达,免疫组织化学法观察HSP70的表达情况,Western blot测定HSP70蛋白的表达水平. 结果 DM组4、8及12周肾脏中HSP70 mRNA及蛋白的表达逐渐升高.同一时间的PT组肾脏HSP70 mRNA及蛋白的表达均较DM组减少（P＜0.05）. 结论 糖尿病大鼠肾脏中HSP70的表达升高,普罗布考可降低HSP70的表达,对肾脏起保护作用.     

热休克条件下诱导型HSP70对人胚肺二倍体细胞衰老影响实验研究

目的 探讨在热休克条件下，人胚肺二倍体细胞MRC-5株诱导型热休克蛋白70（HSP70）在细胞水平的表达机制，诱导型HSP70对MRC-5细胞衰老有无影响及影响MRC-5细胞衰老的相关机制。方法 用热休克处理诱导MRC-5细胞HSP70的表达，同时采用HSP70反义寡核苷酸转染MRC-5细胞作对照，分别置42℃恒温水浴30min、1、2、4h，及每日水浴1h共3和5d，各组细胞均在37℃复温。分别测定细胞HSP70的表达，细胞超氧化物歧化酶（SOD）总活力，并通过衰老相关β-半乳糖苷酶（SA-β-Gal）染色和显微镜观察判断衰老细胞。结果 在42℃热休克处理下，MRC-5细胞HSP70的表达随热处理时间延长而增加（P〈0．05），且均高于同时点对照组水平（P〈0．05）；MRC-5细胞SOD总活力随热处理时间延长而增高，均高于同时间点对照组水平，差异有统计学意义（P〈0．05）；经HSP70反义寡核苷酸处理的细胞，在反复热处理第3天及第5天，SA-β-Gal染色阳性表达率高于实验组。结论 热处理方法可成功诱导MRC-5细胞表达HSP70，随热处理时间延长，MRC-5细胞HSP70表达增加，HSP70可能通过增强细胞内源性抗氧自由基能力，影响MRC-5细胞的衰老进程。     

亚低温对大鼠脑缺血再灌注损伤后HSP70 、TNF-α、NF-κB表达及损伤神经细胞凋亡的影响

目的：观察亚低温对大鼠脑缺血再灌注损伤后HSP70（热休克蛋白70），TNF－α（肿瘤坏死因子－α），NF－κB（核蛋白因子－κB）表达及损伤神经细胞凋亡的影响，方法：采用大鼠局灶性脑缺血再灌注损伤模型，大脑中动脉阻塞2h，再灌注损伤10h，用免疫组织化学法和原位缺口末端标记（TUNEL）法分别检测假手术组，对照组和亚低温组HSP70，TNF－α，NF－κB表达水平和凋亡细胞百分率。结果：亚低温组HSP70表达水平对照组显著升高（P＜0．05），而TNF－α，NF－κB表达水平和凋亡细胞百分率明显低于对照组（P＜0．05），结论：亚低温下调大鼠脑缺血再灌注损伤后TNF－α，NF－κB表达水平和上调HSP70表达水平可能与其抗损伤神经细胞凋亡作用有关。     

辛伐他汀对大鼠急性肺栓塞的保护作用

目的探讨辛伐他汀对大鼠急性肺栓塞的保护作用及其机制。方法将72只Sprague-Dawley大鼠随机分为假手术组,肺栓塞模型组和辛伐他汀干预组,每组24只。在造模后2h,6h,24h分别测定各组大鼠的右心室收缩压（RVSP）和平均肺动脉压（mPAP）,分离肺脏进行HE染色及肺脏病理学检查。在造模后6h测定各组大鼠的动脉血气和肺血管内皮型一氧化氮合酶（eNOS）蛋白表达。应用ELISA法测定各组大鼠血浆白介素-6（IL-6）和肿瘤坏死因子Ⅱ（TNF-α）水平。结果肺栓塞模型组大鼠不同时间点的mPAP,RVSP均较假手术组明显升高（P〈0．01）。辛伐他汀干预组大鼠不同时间点的mPAP、RVSP与肺栓塞模型组比较均显著降低（P〈0．05）。肺栓塞模型组大鼠6h氧分压（PaO2）与假手术组大鼠比较明显降低（P〈0．01）;辛伐他汀干预组大鼠6hPaO2与肺栓塞模型组大鼠比较明显升高（P〈0．05）。肺栓塞模型组大鼠6h肺小动脉eNOS蛋白表达较假手术组降低（P〈0．01）,辛伐他汀干预组组大鼠6h肺小动脉eNOS蛋白表达较肺栓塞模型组明显升高（P〈0．05）。肺栓塞模型组大鼠各时间点血浆IL-6,TNF-α水平较假手术组有升高趋势（P〉0．05）。辛伐他汀干预组大鼠各时间点血浆IL．6、TNF—α水平较肺栓塞模型组大鼠略有降低（P〉0．05）。结论辛伐他汀干预可降低急性肺栓塞大鼠的mPAP和RvsP,减轻低氧血症,改善内皮细胞功能,但对血浆IL-6．TNF-α水平无影响。     

银杏叶提取物对大鼠心肌缺血再灌注损伤热休克蛋白表达的影响

目的:观察热休克蛋白70(HSP70)在心肌缺血再灌注损伤过程中的表达规律及银杏叶提取物(EGB)干预后的变化.方法:65只SD大鼠随机分为假手术组、模型组和治疗组.试验前30 min,治疗组大鼠腹腔注射银杏叶提取物50 mg/kg,结扎左冠状动脉前降支建立大鼠急性心肌缺血再灌注损伤模型.于再灌注不同时间点(1、3、6、12、24、48 h)取出心脏左前降支支配区的全层心肌组织,用免疫组织化学和Western Blot方法观察心肌HSP70的表达.结果:心肌缺血再灌注损伤时HSP70主要表达在心肌细胞和微血管上,表达水平随再灌注时间延长而增加,24 h时达高峰;与模型组相比,治疗组心肌组织HSP70表达水平明显上调,表达时间明显延长.结论:EGB促进HSP70在心肌表达增强和延长,可能与其提高心肌对缺血再灌注损伤的耐受性有关.     

Increased heat shock protein 70 expression in the pancreas of rats with endotoxic shock

AIM: To investigate the ultra-structural changes and heat shock protein 70 (HSP70) expression in the pancreas of rats with endotoxic shock and to detect their possible relationship. METHODS: A total of 33 Wistar rats were randomly divided into three groups: control group (given normal saline), small dose lipopolysaccharide (LPS) group (given LPS 5 mg/kg) and large dose LPS group (given LPS 10 mg/kg). Pancreas was explanted to detect the ultra-structural changes by TEM and the HSP70 expression by immunohistochemistry and Western blot. RESULTS: Rats given small doses of LPS showed swelling and loss of mitochondrial cristae of acinar cells and increased number of autophagic vacuoles in the cytoplasm of acinar cells. Rats given large doses of LPS showed swelling, vacuolization, and obvious myeloid changes of mitochondrial cristae of acinar cells, increased number of autophagic vacuoles in the cytoplasm of acinar cells. HSP70 expression was increased compared to the control group (P<0.05). CONCLUSION: Small doses of LPS may induce stronger expression of HSP70, promote autophagocytosis and ameliorate ultra-structural injuries.     

RNA干扰对大鼠血管内皮细胞血管紧张素转换酶表达的抑制作用

目的用RNA干扰技术选择性下调大鼠血管内皮细胞(ECs)上血管紧张素转换酶(ACE)的表达。方法构建携带ACE基因特异短发夹RNA(shRNA)编码序列的质粒载体pACE-shRNA,与脂质体转染剂制备成不同比例的复合物,转染原代培养的大鼠主动脉ECs,在荧光显微镜下检测转染效率,用四甲基氮唑盐比色法检测细胞存活率,选择出质粒与转染剂的最佳配比。然后以最佳配比复合物转染ECs,分别于转染前及转染后24、48、72h通过半定量RT-PCR和Western blot法检测ACE的表达。结果质粒与转染剂的最佳配比为质粒1.0μg、转染剂4μl,此时转染效率为(81.2±6.2)%,细胞生存率为(90.2±4.1)%。以最佳配比复合物转染ECs后24h,ACE的mRNA和蛋白质表达无明显变化;转染后48h,表达明显降低,差异有统计学意义(P<0.01);72h时表达更低,只有微弱的条带显示。结论RNA干扰能选择性下调原代培养的大鼠血管ECs上ACE的表达,这可能为心血管疾病基因治疗的研究和应用提供新方法。     

小鼠巨细胞病毒感染诱导热休克蛋白70表达的体外实验研究

目的 研究小鼠胚肺组织感染小鼠巨细胞病毒(murine cylomegalovirus,MCMV)后其细胞热休克蛋白70(HSP70)的表达情况,为临床病毒感染性疾病的诊断开辟新途径。方法 体外培养小鼠胚肺细胞,加入MCMV(感染组)后4、8、12、16、24、48、72、96小时分别收获细胞;用免疫组织化学方法检测HSP70表达情况并与对照组(不感染病毒)比较。结果 感染组各时相HSP70表达明显增强,其中96小时表达最强,8小时次之,24小时最弱;随HSP70表达增强细胞变性程度加重。结论 MCMV可诱导小鼠胚肺细胞HSP70的高表达;HSP70表达强度与细胞病变程度呈正相关。     

氟桂利嗪对高血压大鼠脑缺血再灌注后HSP70表达的影响

目的探讨高血压大鼠急性脑缺血再灌注后氟桂利嗪对热休克蛋白70(HSP70)表达的影响.方法将大鼠用双肾动脉狭窄术制成高血压大鼠模型,喂养2个月.随机分为氟桂利嗪组、缺血再灌注组和假手术组.除假手术组大鼠外,其他大鼠均制成大脑中动脉闭塞(MCAO)模型,MCAO时间均为2小时,再灌注时间为1天.经免疫组化染色后用病理图像分析仪测出HSP70阳性细胞数,并计算阳性细胞率.比较三组HSP70阳性细胞率.结果氟桂利嗪组HSP70表达高于缺血再灌注组(P＜0.05).结论氟桂利嗪能促进HSP70表达,对缺血再灌注引起的受损脑组织起保护作用.     

去甲肾上腺素预处理对大鼠供心热休克蛋白70、一氧化氮及一氧化氮合酶表达的影响

目的:探讨去甲肾上腺素预处理是否可诱导心肌热休克蛋白70(HSP70)的合成,并研究其对供心一氧化氮(NO)、一氧化氮合酶(NOS)的影响,探讨去甲肾上腺素预处理心肌保护作用机制。方法:Wistar大鼠18只,分为2组:对照组(C,n=9),腹腔注射0.9%氧化钠注射液0.5 mL,24 h后取离体心脏灌注(Histidine-tryptophan-ketoglutarte,HTK)心脏保护液,4℃保存3 h后建立Langendorff离体心脏灌注模型,灌注(Krebs-Henseleit,K-H)液2 h;实验组(E,n=9)腹腔注射重酒石酸去甲肾上腺素(溶于0.9%氯化钠液中)3.1μmol/kg(0.53 mg/kg),腹腔注射24 h后取离体心脏,处理方法同C组。测定心肌HSP70、NO、NOS的含量以及相关生化指标并做统计学处理比较。结果:HSP70含量E组较C组明显增高(P<0.01),NO、NOS的含量E组较C组明显增多(P<0.01),生化指标E组明显优于C组。结论:去甲肾上腺素预处理能诱导供心心肌组织HSP70、NO、NOS高表达,其对供心具有明显的保护效应,并且其促进心肌NO、NOS的表达,这可能是去甲肾上腺素预处理发挥供心保护作用的机制之一。     

Resistance training increases heat shock protein levels in skeletal muscle of young and old rats

Heat shock proteins (HSP) HSP72, HSC70 and HSP25 protein levels and mRNA levels of HSP72 genes (Hsp72-1, Hsp72-2, Hsp72-3) and HSC70 were examined in tibialis anterior muscles from young and old rats following 4.5 weeks of heavy resistance exercise. Young (3 months) (n=10) and old (30 months) (n=9) rats were subjected to 14 sessions of electrically evoked resistance training using stretch-shortening contractions of the left limb that activated the dorsiflexor muscle group, including the tibialis anterior muscle, while the right side served as the intra-animal control. Muscle wet weight of the left tibialis anterior increased by 15.6% in young animals compared to the untrained right side, while the aged rats demonstrated no significant hypertrophy based on muscle wet weight. There were no differences in mRNA expression between the control and experimental muscles in either the old or the young animals for any of the four genes examined. On the other hand, HSP72 levels as determined by Western blots were significantly (p<0.01) higher (968.8 and 409.1%) in the trained as compared to the contralateral control muscle in young and old animals, respectively. HSP25 expression was increased significantly (p<0.01) by training in muscles of young rats (943.1%) and old rats (420.3%). Moreover, there was no training by age interaction for HSP72, while a significant age and training by age effects were found in muscles for HSP25. There was no change in HSC70 protein expression in response to the training intervention in either age group. SOD-1 enzyme level increased by 66.6% in the trained muscles of the young rats, while this enzyme was 33% lower in trained muscles compared to the untrained control side in old rats. Moreover, a significant (p<0.05) training by age interaction was found for SOD-1 enzyme levels. This study suggests that fast contracting muscles in young and old animals are capable of increasing HSP expression in response to high intensity contractile stress. Furthermore, the data are consistent with the hypothesis that higher levels of oxidative stress in muscles of old animals limit HSP levels and/or function in response to high intensity contractile stress.     

Expression of heat shock protein 70 (HSP70) in patients with colorectal adenocarcinoma--immunohistochemistry and Western blot analysis

The role of heat shock protein 70 (HSP70) expression has been investigated in various types of tumors. There are only little and controversial data about its clinical relevance in colorectal carcinoma, one of the most common carcinomas observed in humans. In this study we investigated expression of HSP70 in human colonic carcinoma and possible correlation with clinicopathology. To assess patterns (cytosolic and membrane) of HSP70 expression, the 48 surgically removed colorectal adenocarcinomas and 12 normal colonic and rectal mucosal samples were examined by immunohistochemistry and Western-blot. According to results of immunohistochemistry, expression of cytoplasmic HSP72 was significantly higher in colorectal carcinoma compared with normal and adjacent mucosa (p<0.01). In addition, there was significant increase in HSP72 expression in lymph node-positive compared to node-negative group (p<0.001). Dukes C2 stage of colonic cancer showed significantly higher immunohistochemical score than Dukes B2 and B1 stage groups (p< 0.05 i.e. p< 0.02). There was no relation between expression of HSP72 and degree of tumor differentiation. Using Western blot analyses, we noticed elevated levels of cytosolic HSP70 in colorectal cancer cells compared to normal. Densitometric analysis of blots of plasma membrane HSP70 expression has shown decrease in colorectal cancer cells compared to normal mucosa. According to our results, overexpression of HSP72 in malignant tissues of patients with colorectal carcinoma is related to tumor progression, suggesting that these proteins could play an important role not only in tumorigenesis but also in the development of drug resistance. Further research is necessary to clarify the mechanisms responsible for differential HSP70 expression as well as its definitive role in colorectal cancer.