痛风宁对急性痛风性关节炎大鼠HSP70表达的影响

目的观察痛风宁对急性痛风性关节炎大鼠滑膜组织热休克蛋白(HSP)70表达的影响。方法将60只Wistar大鼠随机分为5组,每组12只。除空白组外,均采用尿酸钠注射大鼠右踝关节诱发急性痛风性关节炎,药物组分别给予痛风宁高、低剂量,秋水仙碱治疗。给药5 d后,采用HE染色法观察大鼠右踝关节炎症改变,免疫组化LSAB法检测HSP70表达情况。结果与模型组相比,痛风宁治疗组大鼠右踝关节病理切片炎症浸润度、关节肿胀度均明显改善;且大鼠右踝关节滑膜组织HSP70的表达量明显升高。结论痛风宁治疗急性痛风性关节炎明确有效,诱导体内HSP70的高表达是其治疗急性痛风性关节炎的重要作用途径之一。     

基于免疫相关细胞因子研究理冲汤对子宫肌瘤细胞凋亡和增殖的影响

目的：从免疫相关细胞因子表达情况研究理冲汤对子宫肌瘤细胞凋亡和增殖的影响。方法：用异种移植法建立子宫肌瘤小鼠模型,随机分为正常组、模型组、理冲汤组和阳性药组,每组6只,连续药物干预4周;通过透射电子显微镜观察子宫平滑肌细胞的超微结构;采用免疫组织化学染色检测促进细胞凋亡因子胱天蛋白酶3(Caspase-3)、抑制细胞增殖因子Ki67的表达情况;采用MSD电化学发光技术检测白细胞介素-12p70(IL-12p70)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤生长相关基因(GRO/KC)的表达情况。结果：与模型组比较,理冲汤组子宫平滑肌细胞超微结构改善明显,细胞排列紊乱和细胞膜增厚现象均有所减轻;子宫组织中Caspase-3表达升高(P<0.01),Ki67表达降低(P<0.01);血清中IL-12p70表达升高(P<0.05),IL-1β、IL-6、GRO/KC表达降低(P<0.05,P<0.01)。结论：理冲汤具有促进子宫肌瘤细胞凋亡、抑制细胞增殖的作用,其分子机制与调控免疫相关细胞因子表达有关。     

HSP70单克隆抗体诱导宫颈癌细胞凋亡的实验研究

目的:体外培养宫颈癌细胞株,免疫化学方法检测阻断HSP70前后的表达;方法:TUNEL技术观察不同浓度热休克蛋白70单抗对细胞生长的影响.探讨热休克蛋白70(HSP70)单抗对宫颈癌HeLa细胞凋亡的诱导作用;结果:经不同浓度热休克蛋白70单抗后,免疫化学方法检测HSP70的表达显著受到抑制.TUNEL技术检测出细胞凋亡;结论:表明热休克蛋白70单克隆抗体明显诱导宫颈癌细胞调亡.     

Effects of malachite green on the mRNA expression of detoxification‐related genes in Nile tilapia (Oreochromis niloticus) and other major Chinese freshwater fishes

The use of malachite green (MG) in fish farming is prohibited in China due to its potentially toxicological and carcinogenic nature, but it is still illegally used in some places. The aim of this study was to investigate the time and concentration‐dependent responses of xenobiotic metabolizing and detoxification‐related genes in diverse fishes exposed to MG both in vivo and in vitro. Experimental fish were administered to two exposure groups of malachite green (MG) (0.10 and 0.50 mg L^?1) for 8 h. The hepatocytes isolated from Nile tilapia were incubated with MG (0.5, 1.0, and 2.0 mg L^?1) for 8 and 24 h, respectively. In vivo, exposure to 0.10 and 0.50 mg L^?1 MG for 8 h caused significant changes of the detoxification‐related genes on the mRNA expression levels. Low‐concentration (0.10 mg L^?1) level of MG induced significant increase on the mRNA expression level of GSTR gene in Nile tilapia and other fishes. The mRNA expression of grass carp UCP2 was significantly induced when exposed to 0.5 mg L^?1 MG. However, the mRNA expression levels of GSTA, CYP1A, and GPX were inhibited significantly by 0.5 mg L^?1 MG in Nile tilapia, grass carp, and Taiwan snakehead. In vitro, the significant increase of mRNA expression of these genes was detected after exposure to 0.5 mg L^?1 MG (UCP2), and 1.0 mg L^?1 MG (CYP1A1, GSTA, GSTR, and UCP2). The induction of hepatic CYP1A1, GSTA, GSTR, and UCP2 in response to MG suggested a potential role of fish CYP1A1, GSTA, GSTR, and UCP2 in MG metabolism. © 2011 Wiley Periodicals, Inc. Environ Toxicol, 2013.     

E-cadherin和HSP70在肝细胞癌中的表达

目的:探讨E-cadherin和HSP70在不同分化程度HCC中的表达情况以及两者的相关性。方法:采用免疫组织化学方法,检测41例肝癌组织及5例癌旁正常肝脏组织中E-cadherin和HSP70的表达情况。结果:E-cadherin在正常肝脏组织中呈高表达,而在HCC中表达降低且与分化程度有关,部分低分化的HCC组织中表达缺失(P<0.05);HSP70在HCC中表达升高,与正常组比较差异有显著性,且与分化程度有关。相关性分析发现E-cadherin和HSP70在HCC中的表达成负相关(r=-0.322,P<0.05)。结论:HCC中,E-cadherin表达降低,HSP70表达升高,提示两者在HCC侵袭和转移中存在一定的相关性。     

Celastrol regulates innate immunity response via NF-κB and Hsp70 in human retinal pigment epithelial cells

Elevated nuclear factor kappa B (NF-κB) activity and interleukin-6 (IL-6) secretion participates in the pathology of several age and inflammatory-related diseases, including age-related macular degeneration (AMD), in which retinal pigment epithelial cells are the key target. Recent findings reveal that heat shock protein 70 (Hsp70) may affect regulation of NF-κB. In the current study, effects of Hsp70 expression on NF-κB RelA/p65 activity were evaluated in human retinal pigment epithelial cells (ARPE-19) by using celastrol, a novel anti-inflammatory compound. Anti-inflammatory properties of celastrol were determined by measuring expression levels of IL-6 and endogenous NF-κB levels during lipopolysaccharide (LPS) exposure by using enzyme-linked immunosorbent assays (ELISA). Cell viability was measured by MTT and LDH assay, and Hsp70 expression levels were analyzed by Western blotting. ARPE-19 cells were transfected with hsp70 small interfering RNA (siRNA) in order to attenuate Hsp70 expression and activity of NF-κB RelA/p65 was measured using NF-κB consensus bound ELISA.Simultaneous exposures to LPS and celastrol reduced IL-6 expression levels as well as activity of phosphorylated NF-κB at serine 536 (Ser536) in ARPE-19 cells when compared to LPS exposure alone. In addition, inhibition of NF-κB RelA/p65 activity by celastrol was attenuated when Hsp70 response was silenced by siRNA. Favorable anti-inflammatory concentrations of celastrol showed no signs of cytotoxic response. Our findings reveal that celastrol is a novel plant compound which suppresses innate immunity response in human retinal pigment epithelial cells via NF-κB and Hsp70 regulation, and that Hsp70 is a critical regulator of NF-κB.     

慢性肾功能衰竭患者认知功能障碍与血浆HSP70的相关性研究

目的采用蒙特利尔认知评估量表(Montreal CognitiveAssessment,MoCA)评价100例观察慢性肾功能衰竭患者认知功能损害的程度;观察慢性肾功能衰竭患者认知功能的损害程度与血浆HSP70浓度相关性;并同时观察慢性肾功能衰竭伴认知功能障碍患者血浆HSP70浓度与MoCA量表各个子项目的相关性;观察慢性肾功能衰竭伴或不伴认知功能障碍的两组患者(根据MoCA量表评分)与血浆HSP70浓度的相关性。方法运用MoCA量表检测118例慢性肾功能衰竭(chronic renal failure,CRF)患者的认知功能,根据MoCA量表评分标准选择100例CRF患者并将其分为2组,即慢性肾功能衰竭伴认知功能正常组(CRF-NCI,n=50)、和慢性肾功能衰竭伴认知功能障碍组(CRF-CI,n=50)。运用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)测定2组患者血浆HSP70浓度。使用相关性分析观察慢性肾功能衰竭患者认知功能的损害程度与血浆HSP70浓度相关性;采用多元线性回归分析观察慢性肾功能衰竭伴认知功能障碍患者血浆HSP70浓度与MoCA量表各个子项目的相关性;结果 CRF-NCI组和CRF-CI组患者血浆HSP70浓度分别为(10.37±2.86)ng/mL和(17.38±5.38)ng/mL,差异有统计学意义(P<0.01);对HSP70浓度与MoCA量表进行相关性分析显示,慢性肾功能衰竭伴认知功能障碍患者组患者血浆HSP70浓度与MoCA得分呈负相关(r=-0.674,P=0.0001);对慢性肾功能衰竭伴认知功能障碍患者各个认知领域的分值与血浆HSP70浓度水平进行多元线性回归分析显示,血浆HSP70水平与患者记忆能力、视空间执行能力和定向能力存在负相关(R2=0.928,F=78.711,P=0.000)。而与视空间执行能力、命名能力、注意能力和定向能力无明显相关性。结论慢性肾功能衰竭伴认知功能障碍患者HSP70浓度显著高于认知功能正常组。HSP70浓度的改变可以反映慢性肾功能衰竭伴认知功能障碍患者认知功能损害的程度。在慢性肾功能衰竭伴认知功能障碍患者各个认知领域中,血浆HSP70浓度与记忆能力、语言、抽象存在相关性,而与视空间执行能力、命名能力、注意能力和定向能力无相关性。     

神经节苷脂GM1与亚低温对缺氧缺血后脑中HSP70、NPY表达的影响

目的 探讨神经节苷脂GM1与亚低温对缺氧缺血(HI)后新生大鼠脑中热休克蛋白70(HSP70)和神经肽Y(NPY)蛋白和基因表达的影响.方法 50只新生7 d SD大鼠制作HI模型,随机分为假手术(A)组、模型(B)组、神经节苷脂GM1(C)组、亚低温(D)组和神经节苷脂与亚低温联合干预(E)组,每组10只.采用免疫组织化学和逆转录聚合酶链反应(RT-PCR)方法检测大鼠脑中HSP70、NPY蛋白及基因表达.结果 A组HSP70、NPY蛋白及其mRNA表达较弱,B组HSP70蛋白及其 mRNA表达增加,NPY蛋白及其mRNA表达明显增强,各干预组HSP70蛋白和mRNA表达进一步升高,而以E组升高更明显(P＜0.05),而NPY和mRNA在各干预组中表达减弱,联合组减弱更明显(P＜0.05).结论 神经节苷脂GM1与亚低温对新生鼠缺氧缺血性脑损伤(HIBD)的保护作用可能与影响HSP70、NPY的表达有关.     

特拉唑嗪的调脂和抗动脉粥样硬化作用及对热休克蛋白70表达的影响

目的:观察特拉唑嗪对动脉粥样硬化家兔模型的调脂和抗动脉粥样硬化作用及对主动脉管壁热休克蛋白70(HSP70)表达的影响。方法:将27只家兔随机分为正常饮食组、高脂饮食组、特拉唑嗪组。分别试验开始前、喂养后4周、8周、12周测定血浆TG、TC、LDL-C、HDL-C。满12周后处死试验动物,测定主动脉内膜斑块厚度和面积,应用RT-PCR、Western blotting测定主动脉管壁热休克蛋白-70(HSP70)的变化。结果:各组家兔血脂的基线水平差异无显著性。特拉唑嗪显著降低TG、TC、LDL-C(P<0.01),对血清HDL-C水平有升高作用(P<0.05);高脂饮食家兔主动脉内膜的粥样斑块面积和内膜厚度均明显增加,而特拉唑嗪明显抑制了斑块形成和内膜增生。相比于正常饮食组,高脂饮食组家兔主动脉管壁HSP70表达升高(P<0.05),在特拉唑嗪干预之后,HSP70表达升高更为明显(P<0.01)。结论:特拉唑嗪能够降低TC、LDL-C和升高HDL,具有一定的抗动脉粥样硬化作用,其机制除了其降脂作用以外,还可能与增加HSP70表达有关。     

RPL15在K562细胞向红系分化过程中的功能和机制研究

目的研究RPL15基因在K562细胞向红系诱导分化过程中的表达变化,并确定其对珠蛋白表达及红系分化的影响。方法用氯化血红素诱导K562细胞向红系分化,采用实时荧光定量PCR(q PCR)及Western blot法检测RPL15在K562红系分化过程中的表达情况。利用RNA干扰(RNAi)技术抑制K562细胞中RPL15的表达,进而通过四甲基联苯胺染色、q PCR及流式细胞技术检测K562细胞中血红蛋白、红系分化相关基因以及CD235a和CD71表达的变化。结果在K562细胞向红系分化过程中,RPL15的m RNA及蛋白水平均呈先升高后下降趋势。与对照组相比,干扰RPL15表达后,向红系分化24、48和72 h的K562细胞血红蛋白染色阳性率均明显下降;向红系分化的K562细胞中α-、β-、ε-、γ-珠蛋白及红系分化相关基因GATA1和HSP70相对表达量均明显下调(均P<0.05);而抑癌基因P53的m RNA表达水平显著上调(P<0.01)。同时CD235a⁺CD71⁺K562细胞比例在RPL15干扰后显著降低(P<0....