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目的 探讨经人血红素加氧酶1（hHO-1）基因和GATA-4基因联合修饰的大鼠骨髓间充质干细胞（BMSCs）在缺血缺氧环境中抗凋亡及体外分化为心肌细胞样细胞的能力是否优于hHO-1或GATA-4单基因修饰的BMSCs。方法 采用全骨髓贴壁法分离、培养BMSCs并进行鉴定,重组腺病毒转染BMSCs,Western blotting法确定基因表达的最佳时间;以缺氧无血清条件模拟体内缺血缺氧微环境培养基因修饰的BMSCs,采用CCK-8试剂盒和台盼蓝染色法分别检测缺血缺氧培养12、24、48、72h细胞存活率,流式细胞术检测缺血缺氧培养24h细胞凋亡情况,Western blotting及细胞免疫荧光法检测特异性心肌肌钙蛋白I（cTnI）表达情况。结果 缺血缺氧培养12、24、48、72h的hHO-1＋GATA-4组细胞生存率均高于hHO-1组（P〈0.05）和GATA-4组（P〈0.05）。缺血缺氧培养24h的hHO-1＋GATA-4组细胞凋亡率低于hHO-1组（P〈0.05）和GATA-4组（P〈0.05）;GATA-4组与hHO-1＋GATA-组的c Tn I表达差异无统计学意义。结论 hHO-1与GATA-4联合修饰可明显增强缺血缺氧BMSCs的存活;与GATA-4单基因修饰相比,联合修饰并没有增强BMSCs向心肌细胞分化的能力。

目的探讨类叶升麻苷（actesoide）诱导血红素加氧酶-1（HO-1）表达的信号机制。方法类叶升麻苷单独处理体外培养的PC12细胞或运用ERK1/2抑制剂PD98059预处理,提取细胞蛋白,Western blot方法检测pERK1/2和ERK1/2以及HO-1蛋白表达水平。结果与正常对照组相比,类叶升麻苷可以诱导HO-1的表达上调,类叶升麻苷激活ERK1/2,后者激活介导类叶升麻苷诱导HO-1的表达上调,因为PD98059预处理后可以减弱类叶升麻苷引起的HO-1蛋白表达水平降低。结论类叶升麻苷通过激活ERK1/2通路诱导HO-1的表达。

血红素加氧酶-I（HO-1）作为哺乳动物降解血红素的主要途径之一，其主要降解产物一氧化碳（cO）、自由铁（Fe2＋）和胆绿素--在促进细胞生存、细胞内物质循环及免疫调节中具有重要作用。既往研究提示血红素-HO-l通路是决定急性肾损伤（AKI）易感性及严重性的重要内在因素。诱导Ho-l表达能够减轻肾脏缺血一再灌注损伤（IRI）严重程度，而抑制HO-1的表达会加重IRI。本文综述了国内外有关HO．1在AKI诱导保护机制方面的最新研究进展，以便深入认识HO．1在AKI治疗中的作用。

目的观察血红素加氧酶-1（HO-1）对小鼠脾脏T淋巴细胞向CD4＋CD25＋Treg细胞分化的影响及其机制。方法小鼠脾脏淋巴细胞分为3组,分别经PBS、HO-1诱导剂钴原卟啉（Copp,50μmol/L）、HO-1抑制剂锌原卟啉（Znpp,50μmol/L）处理12h,采用实时荧光定量PCR及Western blotting检测各组细胞HO-1 mRNA及蛋白的表达,流式细胞仪检测各组CD4＋CD25＋Treg细胞的比例,ELISA法检测各组细胞上清中Th1、Th2类细胞因子的水平。结果与PBS处理组比较,Copp处理组HO-1 mRNA和蛋白水平均升高,诱导小鼠脾脏T淋巴细胞向CD4＋CD25＋Treg细胞分化的比例增加,Th1类促炎细胞因子表达降低,Th2类抑炎细胞因子表达升高,差异有统计学意义（P〈0.05）,Znpp组则与之相反。结论 HO-1可促进小鼠脾脏T淋巴细胞向CD4＋CD25＋Treg细胞分化,其机制可能与调节Th1/Th2类细胞因子平衡有关。

目的观察重复、长时间正加速度（＋Gz）对动脉粥样硬化家兔模型血管内分泌功能的影响。方法健康雄性新西兰白兔24只,建立动脉粥样硬化模型后随机分为对照组（不进行＋Gz暴露,n=12）和＋Gz暴露组（n=12）。分别于4、8、12周后处死两组动物（每个时间点4只）,采用放射免疫及生物化学法测定静脉血的血管紧张素Ⅱ（AngⅡ）、内皮素（ET）含量及血管内皮细胞的血红素氧化酶-1（HO-1）活力、内源性一氧化碳（CO）浓度及环磷酸鸟苷（cGMP）含量,并对主动脉内膜进行电镜观察。结果随＋Gz暴露时间的延长,实验室兔静脉血AngⅡ、ET及血管内皮细胞HO-1、CO、cGMP含量均显著增高（P〈0.05）,但暴露至第12周时已不再增加;对照组上述指标无明显变化（P=0.05）。对照组动脉内膜下及中膜浅层的泡沫细胞及胶原含量仅见少量增加,而＋Gz暴露8、12周后主动脉内膜下及中膜浅层可见大量增生的泡沫细胞,间质胶原纤维含量明显增加。结论重复、长时间＋Gz暴露会诱发AngⅡ和ET的产生,但也会增加HO-1、CO、cGMP的分泌,对血管免受进一步损伤可能具有潜在的保护作用。

The immunoresponsive gene 1 （IRG 1） protein has crucial functions in embryonic implantation and neurodegeneration. IRG1 promotes endotoxin tolerance by increasing A20 expression in macrophages through reactive oxygen species （ROS）. The cytoprotective protein heme oxygenase-1 （HO-1）, which generates endogenous carbon monoxide （CO）, is expressed in the lung during Lipopolysaccharide （LPS） tolerance and cross tolerance. However, the detailed molecular mechanisms and functional links between IRG1 and HO-1 in the innate immune system remain unknown. In the present study, we found that the CO releasing molecule-2 （CORM-2） and chemical inducers of HO- 1 increased I RG 1 expression in a time- and dose-dependent fashion in RAW264.7 cells. Furthermore, inhibition of HO-1 activity by zinc protoporphyrin IX （ZnPP） and HO-1 siRNA significantly reduced expression of IRG1 under these conditions. In addition, treatment with CO and HO-1 induction significantly increased A20 expression, which was reversed by ZnPP and HO-1 siRNA. LPS-stimulated TNF-a was significantly decreased, whereas IRG1 and A20 were increased by CORM-2 application and HO-1 induction, which in turn were abrogated by ZnPP. Interestingly, siRNA against IRG1 and A20 reversed the effects of CO and HO-1 on LPS-stimulated TNF-a production. Additionally, CO and HO-1 inducers significantly increased IRG1 and A20 expression and downregulated TNF-a production in a LPS-stimulated sepsis mice model. Furthermore, the effects of CO and HO-1 on TNF-α production were significantly reversed when ZnPP was administered. In conclusion, CO and HO-1 induction regulates IRG1 and A20 expression, leading to inhibition of inflammation in vitroand in an in vivomice model.

Gouty arthritis is an inflammatory disease that is caused by an accumulation of monosodium urate （MSU） crystals in the joints. MSU is capable of activating the nucleotide-binding oligomerization domain-like receptor pyrin domain containing 3 （NLRP3） inflammasome, leading to interleukin-lβ （IL-lβ） secretion. Reactive oxygen species （ROS） are major mediators of the NLRP3/IL-lβ interaction. Although nuclear factor E2-related factor 2 （Nrf2） is recognized as a transcription factor that is involved in the response to oxidative stress, the effect of MSU on Nrf2 and on Nrf2-mediated antioxidant enzymes remains unclear. The treatment of THP-1 monocytes using phorbol 12-myristate 13-acetate （PMA） was shown to initiate inflammatory responses. Here, we showed that THP-1 cells, following treatment with MSU crystals, significantly increased IL-1β release, NLRP3 inflammasome activation and ROS production. MSU also promoted the nuclear translocation of Nrf2 and activated lysosomal destabilization. Moreover, the levels of heme oxygenase-1 （HO-1） in gene and protein expressions were upregulated by MSU. MSU-induced IL-lβ secretion and NLRP3 inflammasome activation were inhibited by the knockdown of Nrf2 and via the HO-1 inhibitor zinc （11） protoporphyrin IX （ZnPP）. In addition, HO-1 inhibition increased the level of superoxide anion production and the consumption of glutathione. These findings suggest that Nrf2 and HO-1 mediate redox homeostasis and interact with pro-inflammatory factors in MSU-challenged THP-1 cells, thereby providing new insight into how MSU-induced gouty inflammation is mediated by specific mechanisms that are involved in the Nrf2/Ho-1 antioxidant signaling pathway.

Aim： Our preliminary study shows that a bibenzyl compound isolated from Gastrodia elata, 2-[4-hydroxy-3-（4-hydroxybenzyl）benzyl]-4-（4-hydroxybenzyl）phenol （designated 20C）, protects PC12 cells against H202-induced injury. In this study we investigated whether 20C exerted neuroprotective action in a cell model of Parkinson＇s disease. Methods： A cell model of Parkinson＇s disease was established in PC12 cells by exposure to rotenone （4 pmol/L） for 48 h. Cell viability and apoptosis were assessed, and intracellular ROS level and the mitochondrial membrane potential （MMP） were detected. The expression of apoptosis-related proteins Bax, Bcl-2, cytochrome c, cleaved caspase-3, and oxidative stress-related proteins Nrf2, HO-1 and NQ01 were examined using Western blotting. The mRNA levels of HO-1 and NQ01 were determined with RT-PCR. The nuclear translocation of Nrf2 was observed with immunofluorescence staining. Results： Treatment with rotenone significantly increased the number of apoptotic cells, accompanied by marked increases in the Bax/ Bcl-2 ratio, cytochrome c release and caspase-3 activation. Rotenone also increased ROS accumulation, reduced MMP, and increased the nuclear translocation of Nrf2 as well as the mRNA and protein levels of the Nrf2 downstream target genes HO-1 and NQ01 in PC12 cells. Co-treatment with 20C （0.01-1 pmol/L） dose-dependently attenuated rotenone-induced apoptosis and oxidative stress in PC12 cells. Nrf2 knockdown by siRNA partially reversed the protective effects of 20C in rotenone-treated PC12 cells. Conclusion： The bibenzyl compound 20C protects PC12 cells from rotenone-induced apoptosis, at least in part, via activation of the Nrf2/ARE/HO-1 signaling pathway.

Previous studies have shown that microRNA-1304 （miR-1304） is dysregulated in certain types of cancers, including non-small cell lung cancer （NSCLC）, and might be involved in tumor survival and/or growth. In this study we investigated the direct target of miR-1304 and its function in NSCLC in vitro. Human lung adenocarcinoma cell lines （A549 and NCI-H1975） were studied. The cell proliferation and survival were investigated via cell counting, MTT and colony-formation assays. Cell apoptosis and cell cycle were examined using annexin V-PE/7-AAD and PI staining assays, respectively. The dual-luciferase reporter assay was used to verify post-transcriptional regulation of heme oxygenase-1 （HO-1） by miR-1304. CRISPR/Cas9 was used to deplete endogenous miR-1304. Overexpression of MiR-1304 significantly decreased the number and viability of NSCLC cells and colony formation, and induced cell apoptosis and Go/ G~ phase cell cycle arrest. HO-1 was demonstrated to be a direct target of miR-1304 in NSCLC cells. Restoration of HO-1 expression by hemin （20 IJmol/L） abolished the inhibition of miR-1304 on cell growth and rescued miR-1304-induced apoptosis in A549 cells. Suppression of endogenous miR-1304 with anti-1304 significantly increased HO-1 expression and promoted cell growth and survival in A549 cells. In 17 human NSCLC tissue samples, miR-1304 expression was significantly decreased, while HO-1 expression was significantly increased as compared to normal lung tissues. MicroRNA-1304 is a tumor suppressor and HO-1 is its direct target in NSCLC. The results suggest the potential for miR-1304 as a therapeutic target for NSCLC.

Salidroside,the main active ingredient extracted from Rhodiola crenulata,has been shown to be neuroprotective in ischemic cerebral injury,but the underlying mechanism for this neuroprotection is poorly understood.In the current study,the neuroprotective effect of salidroside on cerebral ischemia-induced oxidative stress and the role of the nuclear factor erythroid 2-related factor 2（Nrf2）pathway was investigated in a rat model of middle cerebral artery occlusion.Salidroside（30 mg/kg）reduced infarct size,improved neurological function and histological changes,increased activity of superoxide dismutase and glutathione-S-transferase,and reduced malon-dialdehyde levels after cerebral ischemia and reperfusion.Furthermore,salidroside apparently increased Nrf2 and heme oxygenase-1 expression.These results suggest that salidroside exerts its neuroprotective effect against cerebral ischemia through anti-oxidant mechanisms and that activation of the Nrf2 pathway is involved.The Nrf2/antioxidant response element pathway may become a new therapeutic target for the treatment of ischemic stroke.