Acute intranasal osteopontin treatment in male rats following TBI increases the number of activated microglia but does not alter lesion characteristics

Intranasal recombinant osteopontin (OPN) has been shown to be neuroprotective in different models of acquired brain injury but has never been tested after traumatic brain injury (TBI). We used a model of moderate-to-severe controlled cortical impact in male adult Sprague Dawley rats and tested our hypothesis that OPN treatment would improve neurological outcomes, lesion and brain tissue characteristics, neuroinflammation, and vascular characteristics at 1 day post-injury. Intranasal OPN administered 1 hr after the TBI did not improve neurological score, lesion volumes, blood–brain barrier, or vascular characteristics. When assessing neuroinflammation, we did not observe any effect of OPN on the astrocyte reactivity but discovered an increased number of activated microglia within the ipsilateral hemisphere. Moreover, we found a correlation between edema and heme oxygenase-1 (HO-1) expression which was decreased in OPN-treated animals, suggesting an effect of OPN on the HO-1 response to injury. Thus, OPN may increase or accelerate the microglial response after TBI, and early response of HO-1 in modulating edema formation may limit the secondary consequences of TBI at later time points. Additional experiments and at longer time points are needed to determine if intranasal OPN could potentially be used as a treatment after TBI where it might be beneficial by activating protective signaling pathways.     

热敏灸治疗脾胃虚弱型糖尿病胃轻瘫疗效观察

目的:观察热敏灸治疗脾胃虚弱型糖尿病胃轻瘫的疗效及相关机制。方法:将70例脾胃虚弱型糖尿病胃轻瘫患者随机分为对照组和观察组各35例。对照组予多潘立酮片,观察组在此基础上予热敏灸治疗,疗程为1个月。治疗前后测定胃轻瘫症状严重指数量表(GCSI)评分、胃排空率,测定患者静脉血血红素加氧酶-1(HO-1)、胰岛素样生长因子-1(IGF-1)水平,于治疗结束后进行疗效评价。结果:观察组总有效率明显高于对照组(P<0.05);治疗后,两组早饱、恶心呕吐、腹胀评分及GCSI总分明显降低(P<0.05),且观察组低于对照组(P<0.05);治疗后,两组胃排空率均明显提升,观察组胃排空率明显高于对照组(P<0.05);观察组HO-1、IGF-1水平在治疗后明显升高,且与对照组有统计学差异(P<0.05)。结论:热敏灸治疗脾胃虚弱型糖尿病胃轻瘫疗效确切,能显著降低GCSI量表评分,改善胃排空率,这可能与其纠正HO-1、IGF-1水平相关。     

The association between inflammatory markers (iNOS,HO-1, IL-33, MIP-1β) and depression with and without posttraumatic stress disorder

Background

Both major depression and posttraumatic stress disorder (PTSD) are characterized by inflammation, increased concentration levels of proinflammatory cytokines, decreased neurogenesis followed by neuroprogression, as well as mitochondrial and the hypothalamic-pituitary-adrenal axis dysfunction. Elevated levels of oxidative stress caused by an increased activity of prooxidants over antioxidants are also observed. Based on several reports, depressive episodes can lead to the sensitization of immune-inflammatory pathways. Thus, depression, PTSD, and depression comorbid with PTSD are associated with immune-inflammatory markers. The study aimed at evaluating concentration levels of iNOS, HO-1, IL-33, and MIP-1β in depression with and without PTSD.

Assessment of ultra-structure,viability and function of lipopolysaccharides-stimulated human dermal fibroblasts treated with chrysin and exosomes (in vitro study)

BackgroundLipopolysaccharides (LPS) stimulate production of inflammatory cytokines. Chrysin is flavonoid beneficial for treatment of inflammatory conditions. Bone marrow mesenchymal stem cell (BM-MSC) exosomes have regenerative ability in different tissues.ObjectiveTo assess potential role of chrysin and BM-MSC exosomes on ultra-structure, viability and function of human dermal fibroblasts-adult (HDFa) stimulated by LPS.MethodsHDFa cells were divided into: Group I: Cells received no treatment. Group II: Cells were stimulated with LPS. Group III: LPS stimulated cells were treated with chrysin. Group IV: LPS stimulated cells were treated with exosomes.ResultsAfter 48 h, ultrastructural examination of HDFa cells in Group I revealed intact plasma membrane and numerous cytoplasmic organelles. Group II displayed destructed plasma membrane and apoptotic bodies. Group III showed intact plasma membrane with loss of its integrity at some areas. Group IV demonstrated intact plasma membrane that showed fusion with exosomes at some areas. Statistical analysis of MTT represented highest mean value of cell viability% in Group IV followed by Groups III, I and II respectively. Statistical analysis of enzyme-linked immunosorbent assay (ELISA) showed the highest mean value of interleukin-1β (IL-1β) was in Group II followed by Groups III, IV and I, while highest mean values of interleukin-10 (IL-10), nuclear factor-erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) proteins were in Group I, followed by Groups IV, III and II respectively.ConclusionsLPS have harmful consequences on ultra-structure, viability and function of HDFa cells. BM-MSC exosomes have better regenerative action on inflamed fibroblasts in comparison to chrysin.     

聚嘧啶束结合蛋白相关剪接因子高表达对糖基化终末产物诱导下视网膜色素上皮细胞损伤的保护作用

目的观察聚嘧啶束结合蛋白相关剪接因子(PSF)高表达对糖基化终末产物(AGEs)诱导下RPE细胞损伤的保护作用。方法将体外培养的人RPE细胞分为正常对照组(N组)、空白对照组(N+AGAGEs组)、空载体对照组(Vec+AGEs组)、PSF高表达组(PSF+AGEs组)。N组RPE细胞常规培养;N+AGEs组只做转染处理但不导入任何外源性基因的RPE细胞联合AGEs诱导;Vec+AGEs组、PSF+AGEs组利用转染试剂脂质体2000将pcDNA空载体或pcDNA-PSF真核表达质粒导入RPE细胞联合AGEs诱导。除N组以外,其余3组细胞进行相应的转染处理,24 h后应用150μg/ml的AGEs刺激72 h。采用HE染色和Hoechst 33258染色观察PSF高表达对RPE细胞凋亡相关形态改变的影响;通过ROS水平检测分析PSF高表达对AGEs诱导的RPE细胞ROS表达的影响;采用MTT比色法检测PSF高表达对RPE细胞生存力的影响;采用Western blot检测PSF不同作用时间及不同剂量对血红素氧合酶1(HO-1)表达的影响。结果HE染色和Hoechst 33258染色观察发现,N组细胞形态饱满,细胞核呈圆形,细胞质丰富,染色均一;N+AGEs组、Vec+AGEs组细胞体积缩小,嗜酸性染色增强,细胞核致密浓染、固缩甚至碎裂;PSF+AGEs组细胞形态尚饱满,细胞浆染色较均匀,细胞核染色均一。MTT比色法检测结果显示,PSF高表达可有效提高RPE细胞生存力,但该作用可被ZnPP有效拮抗,且差异有统计学意义(F=33.26,P<0.05)。DCFH-DA法检测结果显示,与N+AGEs组、Vec+AGEs组比较,PSF+AGEs组细胞中ROS产量下降,差异有统计学意义(F=11.94,P<0.05)。Western blot检测结果显示,PSF蛋白以时间、剂量依赖性的方式上调HO-1的表达水平。PSF蛋白作用24、48、72 h的HO-1相对表达水平较0 h明显升高,差异有统计学意义(F=164.91,P<0.05)。0.1、0.5、1.0、1.5、2.0μg PSF蛋白作用下的HO-1相对表达水平较0.0μg明显升高,差异有统计学意义(F=104.82,P<0.05)。结论PSF可能通过上调HO-1的表达而抑制ROS产生,从而对AGEs诱导下的RPE细胞损伤发挥保护作用。     

Delayed remote preconditioning induces cardioprotection: role of heme oxygenase-1

Background

The role of heme oxygenase-1 (HO-1) in the cardioprotection induced by delayed remote ischemic preconditioning (DRIPC) has not been investigated. Therefore, this study was designed to investigate whether HO-1 is involved in DRIPC-mediated cardioprotection in an isolated perfused rat heart model.

Materials and methods

Isolated rat hearts were subjected to 30 min ischemia followed by 60 min reperfusion. DRIPC (four cycles 5-min occlusion and 5-min reflow at the unilateral hind limb once per day for 1, 2, or 3 d before heart isolation, abbreviated as D1RIPC, D2RIPC, or D3RIPC respectively). Infarct size, myocardial troponin levels, and heart function were measured. The protein and messenger RNA levels of HO-1 were determined.

Results

DRIPC facilitated postischemic cardiac functional recovery and decreased cardiac enzyme release. The infarct size-limiting effect of DRIPC was more pronounced in the D3RIPC group (10.22 ± 2.57%) than the D1RIPC group (22.34 ± 4.02%, P < 0.001) or the D2RIPC group (14.60 ± 3.13%, P = 0.034). These effects in the D1RIPC group could be blocked by Zinc Protoporphyrin IX (ZnPP) (an HO-1 specific inhibitor). DRIPC-mediated cardioprotection was associated with enhanced HO-1 protein expression (D1RIPC, 0.11 ± 0.03; versus 0.15 ± 0.06 in the D2RIPC group, P = 0.06; versus 0.20 ± 0.04 in the D3RIPC group, P = 0.04) and messenger RNA levels of HO-1 expression.

Conclusions

Our findings suggest that HO-1 is involved in the cardioprotection induced by DRIPC, and that increase in the number of preconditioning stimuli may enhance cardioprotective effects accompanied with increased HO-1 level.     

乌司他丁对大鼠原位肝脏移植供肝的保护作用

目的：探讨乌司他丁对大鼠原位肝脏移植供肝的保护作用及机制。 方法：分别用单纯UW液（模型组）或含乌司他丁（乌司他丁组）、HO-1诱导剂CoPP（CoPP组）、HO-1抑制剂ZnPP（ZnPP组）的UW液灌注切取的供体大鼠肝脏并保留灌注液1 h后,原位移植受体大鼠。移植后24 h取移植肝脏与受体大鼠血标本,行肝脏病理学检查及评分;分别用real-time PCR和Western bolt法检测肝组织HO-1 mRNA与蛋白的表达;用Elisa法检测大鼠血清中IL-2和IL-10的含量。 结果：与模型组比较,乌司他丁组与CoPP组供肝的损伤明显减轻、Suzuki评分降低,而ZnPP组损伤加重、Suzuki评分升高（均P<0.05）;乌司他丁组与CoPP组HO-1 mRNA与蛋白的表达明显上调,而ZnPP组明显下调（均P<0.05）;乌司他丁组与CoPP组大鼠血清中IL-2水平明显降低、IL-10水平明显升高,而ZnPP组IL-2水平明显升高、IL-10水平明显降低（均P<0.05）。 结论：乌司他丁可能通过上调移植大鼠肝脏的HO-1水平,减轻再灌注损伤、抑制排斥反应而发挥保护作用。

     

Sufentanil induces delayed myocardial preconditioning mediated by HO-1

Purpose. The objective of this study was to examine whether sufentanil also confers delayed cardioprotection and whether this effect is mediated through HO-1.
Methods. Male Sprague-Dawley rats received either delayed ischemic preconditioning （DIPC） or sufentanilinduced preconditioning （SPC; with 3 μg/kg, 15 μg/kg, 30 μg/kg, 60 μg/kg, or 120 μg/kg sufentanil） or an ischemic reperfusion（CON）. After 24 h, all animals were subjected to a 30 min coronary occlusion followed by a 2 h reperfusion. In the group treated with 120 μg/kg sufentanil, the selective HO-1 inhibitor Zinc protoporphyrin IX （Znpp IX） was administered. The infarct size （IS） was determined with 2,3,5-triphenyltetrazolium chloride staining. Western blotting analysis was used to examine HO-1 expression.
Results. The IS/AAR ratios in the animals treated with DIPC （0.33±0.07） or with SPC （0.44±0.08, 0.32±0.10, 0.32±0.06, and 0.28±0.07 for the groups treated with 15 μg/kg, 30 μg/kg, 60 t~g/kg, or 120 μg/kg sufentanil, respectively） were significantly reduced compared with control （CON） group （0.54±0.06; P〈0.05）. The ED50 of sufentanil was found to be 13.83 μg/kg according to the sigmoid equation. Znpp IX abolished the effect of the 120 μg/kg sufentanil treatment （the IS/ AAR values were 0.54±0.04 for the SPC±Znpp IX group and 0.28±0.07 for the group treated with 120μg/kg SPC; P〈0.05）. The 120 μg/kg SPC treatment increased the expression of HO-1 compared with the CON group（P〈0.05）, and this effect was prevented by Znpp IX （P〈0.05）.
Conclusion. These results indicate that sufentanil produces delayed cardioprotection in anaesthetized rats and HO-1 may be involved in it.     

白细胞介素-10对蛛网膜下腔出血所致脑血管痉挛的治疗作用及机制

目的 观察白细胞介素(IL)-10在蛛网膜下腔出血(SAH)后的脑血管痉挛(CVS)中的治疗作用及其作用机制.方法 日本大耳白兔(30只)随机分为假手术组(A)、SAH组(B)、SAH+IL-10组(C)、SAH+锌原卟啉(ZnPP)+ IL-10组(D)、SAH+ ZnPP组(E),经枕大池2次注血法建立兔SAH后CVS动物模型,术后C、D、E组分别经腹腔注射给予IL-10、ZnPP,术后第5天采用酶联免疫吸附试验(ELISA)法检测血清中肿瘤坏死因子-α(TNF-α)、IL-6的水平,取基底动脉血管检测血管直径并检测血红素氧合酶-1(HO-1)的表达.结果 SAH后血管痉挛模型造模成功,SAH各组基底动脉直径比A组明显缩小(P＜0.05),TNF-α与IL-6含量升高(P＜0.05),其中基底动脉直径在C、D组[(733.94±17.28)、(646.11±9.79)μm]较B、E组[(595.64±10.15)、(532.81±17.09) μm]增加,TNF-α和IL-6含量在C组[(26.27±1.64)、(58.15±1.38) ng/L]、D组[(43.45±1.77)、(77.17±1.09) ng/L]较B组[(53.56±1.27)、(115.93±1.47) ng/L]、E组[(60.56±1.79)、(136.45±1.73) ng/L]降低;A组脑基底动脉未检测到HO-1蛋白,C组HO-1含量(0.446±0.019)较B、D、E(0.314±0.014、0.251±0.018、0.160±0.011)组含量增加.结论 IL-10能够缓解SAH后所致CVS,可能是经HO-1蛋白发挥其治疗作用.     

血红素氧合酶-1与心血管疾病研究

血红素氧合酶-1(HO-1)是血红素氧合酶的诱导型,能催化血红素分解生成CO、胆红素和铁.这三种代谢产物通过多种机制对心血管系统产生保护作用,包括抗氧化应激、抗炎症损伤、抗细胞增殖、抗凋亡、抑制血小板聚集、调节血管张力及参与细胞内信号传递等.HO-1在多种心血管疾病中均有适应性表达,并在防治这些心血管疾病方面发挥重要作用.