硒对小鼠柯萨奇性病毒性心肌炎脂质过氧化反应的影响

目的：探讨硒对小鼠病毒性心肌炎发生中脂质过氧化反应的影响。方法：昆明种小鼠分为常规饲料喂养和适量补硒喂养4周后,腹腔接种柯萨奇B3病毒建立小鼠心肌炎模型,测定小鼠谷胱甘肽过氧化物酶（GSH-Px）、超氧化物歧化酶（SOD）、丙二醛（MDA）含量及心肌病变情况。结果：光镜下补硒组小鼠的心肌病理变化轻,其病理积分明显低于常规喂养组（P〈0.05）。补硒组小鼠GSH-Px、SOD明显高于正常喂养组;MDA明显低于常规喂养组（P〈0.05）。结论：补硒能提高脂质过氧化酶的活性、减少脂质过氧化产物的产生,减轻病毒感染引起的心肌细胞损伤。     

Long non-coding RNA AK085865 ablation confers susceptibility to viral myocarditis by regulating macrophage polarization

Accumulating evidence indicates that regulators of macrophage polarization may exert pivotal functions in the development of coxsackievirus B3 (CVB3)-induced viral myocarditis (VM). However, the mechanisms underlying macrophage polarization remain to be explored. Here, we sought to identify novel and functionally important long non-coding RNAs (lncRNAs) during macrophage polarization and to investigate their function and contribution to VM. In this study, we identified the lncRNA AK085865 as an important regulator of macrophage polarization. Knock-down of AK085865 diminished phenotypical expression of M2 macrophages while promoting polarization to the M1 phenotype. Moreover, AK085865^−/− mice had increased susceptibility to CVB3-induced VM. We observed striking bias towards M1 macrophages, whereas the M2 population was decreased in AK085865^−/− VM mice. Collectively, our findings uncover a critical role of AK085865 in the regulation of macrophage polarization in vitro and in vivo, identifying a new player in the development of VM and providing a potential clinically significant therapeutic target.     

非编码RNA与病毒性心肌炎

病毒性心肌炎(Viral myocarditis,VMC)是一种由病毒感染所引起的以心肌细胞炎症为特征的疾病。由于病毒性心肌炎的发病机制尚未完全研究清楚,因此该病的诊断及治疗对于临床医生来说仍具有极大的挑战性。非编码RNAs (Non-coding RNAs,ncRNAs)是一类不具有编码蛋白质功能的RNA,越来越多的研究表明ncRNAs参与到调控VMC的发生和发展过程中,这可能成为VMC的治疗或诊断的新研究靶点。文中对近3年来关于ncRNAs在VMC的发病机制及诊断中可能发挥的作用进行了综述。     

Studies on the relations of selenium and Keshan disease

Keshan disease is an endemic cardiomyopathy of unknown cause in The People’s Republic of China that occurs most frequently in children under 15 years of age and women of child-bearing age. Studies of children 1–9 years old in Mianing County of Sichuan Province have indicated that Keshan disease is a selenium responsive condition. Incidence rates of 9.5–13.5/1000 in 1974–1975 were reduced to 1–2/1000 in children treated with a tablet weekly of 0.5–1 mg sodium selenite. During 1974–1977, only 21 cases of the disease occurred in 36,603 treated children, compared with 106 cases in 9430 untreated children, of whom 53 died and 5 still have insufficient heart function. Occurrence of the disease was invariably associated with a lower selenium content of cereals, and of hair (less than 0.12 ppm Se) in residents from affected, compared with non-affected, areas. The dose relationship between selenium and regional characteristics of Keshan disease suggests that it is probably a biogeochemical disease; other etiological factors have also been considered.     

Stream ecosystem response to, and recovery from, experimental exposure to selenium

The effects of selenium on streamecosystems were studied in outdoor,experimental stream mesocosms during a dosingperiod in which sodium selenite was added atnominal concentrations of 30 µg/L,10 µg/L, and 2.5 µg/L. The durationof the high, medium, and low treatments were573 d, 972 d, and 311 d, respectively. Apost-dosing period of three years (hightreatment) and two years (medium, lowtreatments) also was studied. Seleniumconcentrations in water, sediment, plants, andmacroinvertebrates were measured throughoutthe dosing and recovery periods. Fatheadminnows and bluegill sunfish were periodicallyheld in the streams to measure seleniumaccumulation and its effects on fish survivaland reproduction. Quantitative samples ofmacroinvertebrates were collected to assessselenium effects on macroinvertebratecommunities.Mean selenium concentration inwater was quite close to the nominalconcentration. Selenium accumulated in thesediment in all three treated streams, but notin the control streams. Sediment seleniumdecreased slowly after dosing ceased, but wasstill significantly higher than in controlstreams three years (high treatment) and twoyears (medium treatment) later.Macrophytetissue selenium concentrations weresignificantly greater in all three treatmentsthan those in the control streams duringdosing. Macrophyte selenium bioaccumulationfactors (BAFs) ranged from about 300 to 1900. Tissue selenium decreased rapidly in all threetreatments after dosing ended.During dosing,selenium concentrations in animals from allthree treatments were significantly higherthan in those from control streams. The BAFsfor macroinvertebrates ranged from 1100 to2000. Isopods accumulated more, and amphipodsless, selenium than other invertebrates. Therewere no significant effects of selenium onmacroinvertebrate abundance, richness ordiversity. Several macroinvertebrates werenot affected by exposure to selenium, butisopod and Tubifex populations weredramatically reduced in the high and mediumtreatments. After dosing, mean seleniumconcentration in macroinvertebrates decreasedslowly.Bluegill sunfish accumulated seleniumduring dosing and after selenium additionsceased. Tissue selenium was highest in theliver, followed by the gonads, skeletalmuscle, and whole body. Tissue seleniumconcentrations one (high, medium) and two(high) years after dosing were lower thanduring dosing, but whole body, skeletal muscleand liver concentrations were high enough tobe considered potentially toxic.Recovery ofselenium contaminated streams includes bothreduction of tissue selenium concentration tonon-toxic levels in fish and their foodorganisms and recovery of populations of taxadeleteriously affected by selenium exposure. Our results suggest that when selenium iseliminated from the water in streams, seleniumconcentrations in sediment, plants,macroinvertebrates, and fishes will decreaseto levels that approach concentrationsconsidered to be non-toxic to fish andwildlife and that affected populations willrecover within several years. Based onselenium accumulation in the food chain andthe presence of real, but not statisticallysignificant, effects on fish mortality andreproduction in the low treatment streams, wesupport a selenium water quality criterion forthe protection of fishes and sensitiveinvertebrates of 2 µg/L or less.     

Truncated monocyte chemoattractant protein‐1 can alleviate cardiac injury in mice with viral myocarditis via infiltration of mononuclear cells

BALB/c mice inoculated intraperitoneally with coxsackievirus group B type 3 (CVB3) were allocated to five groups; namely, a viral myocarditis group infected with CVB3 alone (control group), an antibody intervention group that received intracardiac anti‐MCP‐1, an antibody intervention control group that received goat IgG, a tMCP‐1 intervention group that received plasmid pVMt expressing tMCP‐1, and a tMCP‐1 intervention control group that received plasmid pVAX1. There was also a normal control group. The ratio of murine heart weight to body weight, pathological score of myocardial tissue, serum creatine kinase‐MB titers and CVB3 loading of myocardial tissue were assessed. The cardiac lesions in mice that received 20, 40 or 60 µg pVMt (P < 0.05) were less severe than those in control mice with untreated viral myocarditis. In addition, fewer mononuclear cells had infiltrated the myocardium of mice who received 40 or 60 µg pVMt intramyocardially (P < 0.01), whereas there was no difference in mononuclear cell infiltration between mice with viral myocarditis and those that received 20 µg pVMt (P > 0.05). There was also no difference between mice that received anti‐MCP‐1 antibody and those that received 40 µg pVMt in ratio of HW/BW, serum CK‐MB titers and pathological score (P > 0.05). This study showed that tMCP‐1 can alleviate cardiac lesions and cardiac injury in mice with viral myocarditis via infiltration of mononuclear cells. Thus, tMCP‐1 may be an alternative to anti‐MCP‐1 antibody treatment of viral myocarditis. Further research is required.     

Interacting nutritional and infectious etiologies of Keshan disease

In 1979, Chinese scientists reported that selenium had been linked to Keshan disease, an endemic juvenile cardiomyopathy found in China. However, certain epidemiological features of the disease could not be explained solely on the basis of inadequate selenium nutrition. Fluctuations in the seasonal incidence of the disease suggested involvement of an infectious agent. Indeed, a coxsackievirus B4 isolated from a Keshan disease victim caused more heart muscle damage when inoculated into selenium-deficient mice than when given to selenium-adequate mice. Those results led us to study the relationship of nutritional status to viral virulence. Coxsackievirus B3/0 (CVB3/0), did not cause disease when inoculated into mice fed adequate levels of Se and vitamin E. However, mice fed diets deficient in either Se or vitamin E developed heart lesions when infected with CVB3/0. To determine if the change in viral phenotype was maintained, we passaged virus isolated from Se-deficient hosts, maintained, we passaged virus isolated from Se-deficient hosts, designated as CVB3/0 Se-, back into Se-adequate hosts. The CVB3/0 Se- virus caused disease in Se-adequate mice. To determine if the phenotype change was due to changes in the viral genome, we sequenced viruses isolated from Se-deficient mice and compared them with the input CVB3/0 virus. Six point mutations differed between the parent strain and the recovered CVB3/0 Se- isolates. When the experiment was repeated using vitamin E-deficient mice, the same 6 point mutations were found. This is the first report of a specific host nutritional deficiency altering viral genotype. Keshan disease may be the result of several interacting causes including a dominant nutritional deficiency (selenium), other nutritional factors (vitamin E, polyunsaturated fatty acids), and an infectious agent (virus).     

Exosomes derived from umbilical cord mesenchymal stem cells alleviate viral myocarditis through activating AMPK/mTOR‐mediated autophagy flux pathway

Human umbilical cord mesenchymal stem cell‐derived exosomes (hucMSC‐exosomes) have been implicated as a novel therapeutic approach for tissue injury repair and regeneration, but the effects of hucMSC‐exosomes on coxsackievirus B3 (CVB3)‐induced myocarditis remain unknown. The object of the present study is to investigate whether hucMSC‐exosomes have therapeutic effects on CVB3‐induced myocarditis (VMC). HucMSC‐exosomes were identified using nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM) and Western blot. The purified hucMSC‐exosomes tagged with PKH26 were tail intravenously injected into VMC model mice in vivo and used to administrate CVB3‐infected human cardiomyocytes (HCMs) in vitro, respectively. The effects of hucMSC‐exosomes on myocardial pathology injury, proinflammatory cytokines and cardiac function were evaluated through haematoxylin and eosin (H&E) staining, quantitative polymerase chain reaction (qPCR) and Doppler echocardiography. The anti‐apoptosis role and potential mechanism of hucMSC‐exosomes were explored using TUNEL staining, flow cytometry, immunohistochemistry, Ad‐mRFP‐GFP‐LC3 transduction and Western blot. In vivo results showed that hucMSC‐exosomes (50 μg iv) significantly alleviated myocardium injury, shrank the production of proinflammatory cytokines and improved cardiac function. Moreover, in vitro data showed that hucMSC‐exosomes (50 μg/mL) inhibited the apoptosis of CVB3‐infected HCM through increasing pAMPK/AMPK ratio and up‐regulating autophagy proteins LC3II/I, BECLIN‐1 and anti‐apoptosis protein BCL‐2 as well as decreasing pmTOR/mTOR ratio, promoting the degradation of autophagy flux protein P62 and down‐regulating apoptosis protein BAX. In conclusion, hucMSC‐exosomes could alleviate CVB3‐induced myocarditis via activating AMPK/mTOR‐mediated autophagy flux pathway to attenuate cardiomyocyte apoptosis, which will be benefit for MSC‐exosome therapy of myocarditis in the future.     

Producing selenium-enriched eggs and meat to improve the selenium status of the general population

The role of selenium (Se) in human health and diseases has been discussed in detail in several recent reviews, with the main conclusion being that selenium deficiency is recognised as a global problem which urgently needs resolution. Since selenium content in plant-based food depends on its availability from soil, the level of this element in food and feeds varies among regions. In general, eggs and meat are considered to be good sources of selenium in human diet. When considering ways to improve human selenium intake, there are several potential options. These include direct supplementation, soil fertilisation and supplementation of food staples such as flour, and production of functional foods. Analysing recent publications related to functional food production, it is evident that selenium-enriched eggs can be used as an important delivery system of this trace mineral for humans. In particular, developments and commercialisation of organic forms of selenium have initiated a new era in the availability of selenium-enriched products. It has been shown that egg selenium content can easily be manipulated to give increased levels, especially when organic selenium is included in hen’s diet at levels that provide 0.3–0.5 mg/kg selenium in the feed. As a result, technology for the production of eggs delivering ≈50% (30–35 μg) of the human selenium RDA have been developed and successfully tested. Currently companies all over the world market selenium-enriched eggs including the UK, Ireland, Mexico, Columbia, Malaysia, Thailand, Australia, Turkey, Russia and the Ukraine. Prices for enriched eggs vary from country to country, typically being similar to free-range eggs. Selenium-enriched chicken, pork and beef can also be produced when using organic selenium in the diet of poultry and farm animals. The scientific, technological and other advantages and limitations of producing designer/modified eggs as functional foods are discussed in this review.     

Low‐intensity pulsed ultrasound attenuates cardiac inflammation of CVB3‐induced viral myocarditis via regulation of caveolin‐1 and MAPK pathways

The aggressive immunological activity elicited by acute viral myocarditis contributes to a large amount of cardiomyocytes loss and poor prognosis of patients in clinic. Low‐intensity pulsed ultrasound (LIPUS), which is an effective treatment modality for osteoarthropathy, has been recently illustrated regulating the overactive inflammatory response in various diseases. Here, we aimed to investigate whether LIPUS could attenuate coxsackievirus B3 (CVB3) infection‐induced injury by coordinating the inflammatory response. Male BALB/c mice were inoculated intraperitoneally with CVB3 to establish the model of acute viral myocarditis. LIPUS treatment was given on Day 1, Day 1, 3 and Day 1, 3, 5 post‐inoculation, respectively. All mice were followed up for 14 days. Day 1, 3, 5 LIPUS treatment significantly improved the survival rate, attenuated the ventricular dysfunction and ameliorated the cardiac histopathological injury of CVB3‐infected mice. Western blotting analysis showed Day 1, 3, 5 LIPUS treatment decreased pro‐inflammatory cytokines, increased the activation of caveolin‐1 and suppressed p38 mitogen‐activated protein kinase (MAPK) and extracellular signal‐regulated kinase (ERK) signallings in heart tissue. RAW264.7 cells were treated with lipopolysaccharides (LPS) to simulate the augmented inflammatory response in vivo. LIPUS treatment on RAW264.7 inhibited the expression of pro‐inflammatory cytokines, activated caveolin‐1 and suppressed p38 MAPK and ERK signallings. Transfecting RAW264.7 with caveolin‐1 siRNA blunted the suppression of pro‐inflammatory cytokines and MAPK signallings by LIPUS treatment. Taken together, we demonstrated for the first time that LIPUS treatment attenuated the aggressive inflammatory response during acute viral myocarditis. The underlying mechanism may be activating caveolin‐1 and suppressing MAPK signallings.     

Differential susceptibility of cells expressing allogeneic MHC or viral antigen to killing by antigen-specific CTL

CD8(+) cytotoxic T lymphocytes (CTLs) generated by immunization with allogeneic cells or viral infection are able to lyse allogeneic or virally infected in vitro cells (e.g., lymphoma and mastocytoma). In contrast, it is reported that CD8(+) T cells are not essential for allograft rejection (e.g., heart and skin), and that clearance of influenza or the Sendai virus from virus-infected respiratory epithelium is normal or only slightly delayed after a primary viral challenge of CD8-knockout mice. To address this controversy, we generated H-2(d)-specific CD8(+) CTLs by a mixed lymphocyte culture and examined the susceptibility of a panel of H-2(d) cells to CTL lysis. KLN205 squamous cell carcinoma, Meth A fibrosarcoma, and BALB/c skin components were found to be resistant to CTL-mediated lysis. This resistance did not appear to be related to a reduced expression of MHC class I molecules, and all these cells could block the recognition of H-2(d) targets by CTLs in cold target inhibition assays. We extended our observation by persistently infecting the same panel of cell lines with defective-interfering Sendai virus particles. The Meth A and KLN205 lines infected with a variant Sendai virus were resistant to lysis by Sendai virus-specific CTLs. The Sendai virus-infected Meth A and KLN205 lines were able to block the lysis of Sendai virus-infected targets by CTLs in cold target inhibition assays. Taken together, these results suggest that not all in vivo tissues may be sensitive to CTL lysis.     

Effects of nationwide addition of selenium to fertilizers on foods,and animal and human health in Finland: From deficiency to optimal selenium status of the population

Despite different geological features the Nordic countries are generally selenium-poor areas. In each country various factors such as food importation and life-style determine the selenium (Se) intake. Due to an extremely low Se intake in the 1970s in Finland, 0.025 mg/day, an official decision was made in 1984 to supplement multinutrient fertilizers with Se in the chemical form of sodium selenate. Almost all fertilizers used in Finland since 1985 have contained Se. Currently all crop fertilizers contain 15 mg Se/kg. Finland is still the only country to take this country-wide measure.In a national monitoring programme, sampling of cereals, basic foodstuffs, feeds, fertilizers, soils, and human tissues has been carried out annually since 1985 by four governmental research organizations. Sampling of foods has been done four times per year and human blood has been obtained annually from the same (n = 60) adults. The accuracy of analyses has been verified by annual interlaboratory quality control. During this programme the selenium concentration of spring cereals has increased on average 15-fold compared with the level before the Se fertilization. The mean increase in the Se concentration in beef, pork and milk was 6-, 2- and 3-fold. In terms of Se, organically grown foods of plant origin are generally comparable to products produced before the Se supplementation of fertilizers. Milk from organically fed cows is 50% lower in Se than the usual milk. The average dietary human intake increased from 0.04 mg Se/day/10 MJ in 1985 to a present plateau of 0.08 mg Se/day/10 MJ, which is well above the current nutrition recommendations. Foods of animal origin contribute over 70% of the total daily Se intake. The mean human plasma Se concentration increased from 0.89 μmol/L to a general level of 1.40 μmol/L that can be considered to be an optimal status. The absence of Se deficiency diseases and a reference population have made conclusions on the impact on human health difficult. However, the rates of cardiovascular diseases and cancers have remained similar during the pre- and post-supplementation indicating medical and life-style factors to be much stronger determinants than Se. The nationwide supplementation of fertilizers with sodium selenate is shown to be effective and safe in increasing the Se intake of the whole population. Also, the health of animals has improved.     

病毒感染引起的炎症反应：宿主对抗病毒的“双刃剑”

先天性免疫系统作为宿主抵抗外来病原入侵的第一道防线,也是最迅速的防御系统。宿主先天性免疫系统中的模式识别受体识别入侵信号并激活炎症信号通路,诱导产生大量促炎性细胞因子,引起炎症反应。病毒感染是激活炎症反应的条件之一,诱导机体产生强烈的免疫应答,强大的炎症反应调控网络在宿主抗病毒过程中发挥关键作用,以维持机体的平衡。本文综述了病毒感染引起的炎症反应,重点介绍了宿主对炎症反应的调控网络,以及DNA和RNA病毒对炎症反应的调节机制,为病毒感染引起的免疫性疾病的治疗提供参考。     

Clinical aspects of selenium metabolism

Whereas defined selenium deficiency diseases are well characterized in animals, analogous syndromes in humans are unknown or occur only rarely under conditions of extreme selenium depletion. However, selenium deficiency has since been recognized to play important secondary roles in a variety of human diseases, and several of these can be prevented or treated by means of selenium supplementation. In clinical practice, the selenium status of patients should be monitored routinely, and corrective measures should be instituted to replete the Se body stores of patients where necessary. Interactions of Se with other elements are also of potentially great clinical significance. The importance of such interactions is exemplified by the effects of cadmium on selenium metabolism in spontaneously hypertensive rats. Deceased.     

Angiotensin II–C–C chemokine receptor2/5 axis‐dependent monocyte/macrophage recruitment contributes to progression of experimental autoimmune myocarditis

Angiotensin II (ANG II) plays critical roles in modulation of circulatory homeostasis and activation of innate and adaptive immunity and has also been implicated in several mouse models of autoimmune disease. However, how ANG II regulates macrophages and is involved in development of experimental autoimmune myocarditis (EAM) remains unclear. Therefore, the present study aimed to address the above question and explore possible mechanisms. EAM was induced in BALB/c mice. ANG II was quantitated by ELISA and hematoxylin and eosin staining was employed to analyze pathological changes and macrophage infiltration. The chemotactic ability of ANG II was assessed by using a Transwell system. It was found that ANG II is up‐regulated in serum and heart tissues of mice with EAM and that ANG II significantly drives monocyte/macrophage infiltration through the C‐C chemokine receptor 2/5 (CCR2/5) axis. CCR2/5 antagonists and ANG II receptor inhibitor could all abrogate monocyte/macrophage infiltration and ameliorate development of EAM. Our results have firstly identified a novel function of ANG II: that it is a critical chemokine for monocyte/macrophage recruitment. Furthermore, our results indicate that ANG II is a potential candidate for treatment of inflammatory diseases.

     

HLA-DR53 molecules are associated with susceptibility to celiac disease and selectively bind gliadin-derived peptides

 Celiac disease (CD) patients usually express a DQ2 heterodimer, whose chains DQα1*0501/DQβ1*0201, are encoded by the genes HLA-DQA1*0501 and DQB1*0201, respectively. Among the DQ2 carriers, the risk of developing disease was shown to correlate with the number of DQβ1*0201 chains encoded. Studying two separate cohorts of Italian and Tunisian patients, we now show a significant association of celiac disease with expression of either the DQ2 or DR53 heterodimers. The risk is maximal for individuals that carry both DQ2 and DR53 heterodimers. When twenty synthetic peptides overlapping most of A-gliadin sequence were tested for the binding to various purified DR molecules, it was found that DR53 molecules bind selectively and with high affinity (IC50<1 μM) to A-gliadin-derived peptides. These data suggest that both HLA DQ2 and DR53 molecules are associated with increased genetic risk for CD, and provide a possible biochemical basis for this complex association. Received: 1 August 1998 / Revised: 24 February 1999     

Carvedilol has stronger anti-inflammation and anti-virus effects than metoprolol in murine model with coxsackievirus B3-induced viral myocarditis

Aims

This study aims to compare the effects of carvedilol and metoprolol in alleviating viral myocarditis (VMC) induced by coxsackievirus B3 (CVB3) in mice.

Methods

A total of 116 Balb/c mice were included in this study. Ninety-six mice were inoculated intraperitoneally with CVB3 to induce VMC. The CVB3 inoculated mice were evenly divided into myocarditis group (n = 32), carvedilol group (n = 32) and metoprolol group (n = 32). Twenty mice (control group) were inoculated intraperitoneally with normal saline. Hematoxylin and eosin staining and histopathologic scoring were used to investigate the effects of carvedilol and metoprolol on myocardial histopathologic changes on days 3 and 5. In addition, serum cTn-I levels, cytokine levels and virus titers were determined using chemiluminescence immunoassay, enzyme-linked immunosorbent assay and plaque assay, respectively, on days 3 and 5. Finally, the levels of phosphorylated p38MAPK were studied using immunohistochemical staining and Western blotting on day 5.

Results

Carvedilol had a stronger effect than metoprolol in reducing the pathological scores of VMC induced by CVB3. Both carvedilol and metoprolol reduced the levels of cTn-I, but the effect of carvedilol was stronger. Carvedilol and metoprolol decreased the levels of myocardial pro-inflammatory cytokines and increased the expression of anti-inflammatory cytokine, with the effects of carvedilol being stronger than those of metoprolol. Carvedilol had a stronger effect in reducing myocardial virus concentration compared with metoprolol. Carvedilol was stronger than metoprolol in decreasing the levels of myocardial phosphorylated p38MAPK.

Conclusions

In conclusion, carvedilol was more potent than metoprolol in ameliorating myocardial lesions in VMC, probably due to its stronger modulation of the balance between pro- and anti-inflammatory cytokines by inhibiting the activation of p38MAPK pathway through β1- and β2-adrenoreceptors.     

Targeting UBE4A Revives Viperin Protein in Epithelium to Enhance Host Antiviral Defense

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