Heme Oxygenase-1 Expression in Rats with Acute Lung Rejection and Implication

This study investigated the expression of hemeoxygenase-1 （HO-1） in rats with acute lung rejection and its implication. A valid rat orthotopic left lung transplantation model （SD rat→Wistar rat） was established by using an improved three-cuff anastomosis technique. The rats were divided into control group, CoPP （HO-1 inducer）-treated group and ZnPP （HO-1 inhibitor）-treated group. The severity of acute rejection was graded on the basis of the morphologic changes of the lung samples stained with HE. The expression of HO-1 protein in lung tissue was detected by using immunohistochemistry and Western blot, and HO-1 mRNA activity was assayed by RT-PCR. The results showed that the expression of HO-1 protein was significantly increased with the acute rejection grading in rats （P〈0.01）. As compared with control and ZnPP-treated groups, the severity of acute rejection was not alleviated and the grade not reduced significantly in CoPP-treated group （P〉0.05）. It was concluded that HO-1 protein might be involved in the pathological process of post-graft acute rejection. The expression of HO-1 protein was increased gradually with aggravation of acute rejection, and HO-1 protein might be used as an index to monitor acute rejection after lung transplantation.

靛蓝对脂多糖诱导的THP-1细胞的抗炎作用

目的研究靛蓝(indigo,IDG)对脂多糖(lipopolysaccharide,LPS)致人巨噬细胞(THP-1来源)炎症损伤的影响。方法建立LPS诱导THP-1细胞炎症损伤模型,分别用0.4、4、40μmol/L的靛蓝作用于THP-1细胞炎症损伤模型1 h,实时荧光定量PCR(qRT-PCR)法检测白介素(IL)-1β、IL-6、IL-8、IL-23p19和肿瘤坏死因子(tumor necrosis factor,TNF)-α的mRNA表达的变化;ELISA实验检测细胞培养上清中IL-1β、IL-6、IL-8、IL-23p19和TNF-α的含量差异。Western blot实验检测核因子E2相关因子2(nuclear factor E2-related factor 2,Nrf2)和血红素氧合酶1(heme oxygenase-1,HO-1)蛋白水平的影响。结果与模型组比较,靛蓝能够显著抑制LPS诱导的细胞IL-1β、IL-6、IL-8、IL-23p19和TNF-α的转录水平(P<0.05)以及细胞培养上清中IL-1β、IL-6、IL-8、IL-23p19和TNF-α的含量(P<0.05);同时能够显著增加Nrf2和HO-1的表达(P<0.05)。结论靛蓝可抑制LPS引起的THP-1细胞炎症反应,其抗炎机制可能与激活Nrf2/HO-1通路有关。

Protective effect of heme oxygenase-1 on lung injury induced by erythrocyte instillation in rats

Background Intratracheal instillation of blood induces self-repaired acute lung injury. However, the mechanism of repair has been unclear. Heme-oxygenase （HO）-1, which catalyzes heme breakdown, acts as an inducible defense against oxidative stress and plays an important role in inflammation. The objective of this study was to test the role of HO-1 in lung injury caused by intratracheal instillation of red cells. Methods Forty healthy, male Sprague-Dawley rats were randomly divided into five groups： normal group, saline group, erythrocyte group, erythrocyte＋zinc-protoporphyrin （ZnPP, HO-1 inhibitor） group and saline＋ZnPP group. At 2 days after intratracheal instillation of red cells, lung tissues and lavage samples were isolated for biochemical determinations and histological measurements. Results Histological analysis revealed that administration of ZnPP worsened the acute lung injury induced by instilled erythrocytes. HO-1 was over-expressed in the erythrocyte group and in the erythrocyte ＋ ZnPP group. Compared with the erythrocyte ＋ ZnPP group, the levels of total protein, lactate dehydrogenase and tumor necrosis factor-a in the lavage were lower （P 〈0.01）, while the level of interleukin-10 was higher in the erythrocyte group （P 〈0.01）. Conclusion HO-1 protects against erythrocyte-induced inflammatory injury in lung.

Piperlongumine Inhibits Zika Virus Replication In vitro and Promotes Up-Regulation of HO-1 Expression, Suggesting An Implication of Oxidative Stress

Owing to the widespread distribution of mosquitoes capable of transmitting Zika virus, lack of clinical vaccines and treatments, and poor immunity of populations to new infectious diseases, Zika virus has become a global public health concern. Recent studies have found that Zika virus can continuously infect human brain microvascular endothelial cells.These cells are the primary components of the blood–brain barrier of the cerebral cortex, and further infection of brain tissue may cause severe damage such as encephalitis and fetal pituitary disease. The present study found that a biologically active base, piperlongumine(PL), inhibited Zika virus replication in human brain microvascular endothelial cells, Vero cells, and human umbilical vein endothelial cells. PL also significantly increased heme oxygenase-1(HO-1) gene expression, while silencing HO-1 expression and using the reactive oxygen species scavenger, N-acetylcysteine, attenuated the inhibitory effect of PL on Zika virus replication. These results suggest that PL induces oxidative stress in cells by increasing reactive oxygen species. This, in turn, induces an increase in HO-1 expression, thereby inhibiting Zika virus replication. These findings provide novel clues for drug research on the prevention and treatment of Zika virus.

Effect of TLR-4 and HO-1 on acute lung injury induced by hemorrhagic shock in mice

Objective： To examine whether TLR-4 has an ettect on hemorrhage induced changes in lung, and to investigate the change of heme oxygenase-1 （HO-1） on acute lung injury （ALl） induced by hemorrhagic shock in mice. Methods： Forty-eight male mice, including C3H/HeN mice and C3H/HeJ mice, were randomly divided into sham group （n=12）, hemorrhagic shock group with twelve mice in each phase. Blood pressure （BP） was monitored continuously by attaching carotid artery catheter to a strain gauge pressure transducer/ polygraph. Arterial blood samples were taken for blood gas analysis. A mouse model of non-lethal hemorrhagic shock and resuscitation was used to observe pulmonary myeloperoxidase （MPO） activity and wet/dry weight ratio （W/D）. The expression of HO- 1 was observed by means of RT-PCR and immunohistochemistry. IL-6 and IL-10 in lung tissue homogenate were assayed by enzyme-linked immunosorbent assay （ELISA）. The pulmo- nary pathologic changes were observed under electron microscope and light microscope. Results： Compared with sham group, the expression of HO- 1 in lung tissue was significantly higher in Hem 24 h and Hem 48 h of C3H/HeN mice （P〈0.01）. The expression of HO-1 mRNA and the levels of IL-6, IL-10 and MPO in lung tissue were markedly increased in Hem 24 h （P〈0.01 or P〈0.05）; Compared with C3H/HeN mice, the expression of HO- 1 rnRNA and the levels of IL-6 and IL-10 in C3H/HeJ mice significantly decreased in Hem 24 h and Hem 48 h （P〈0.01 or P〈O.05）, and the W/D, MPO in C3H/HeJ mice were obvi- ously lower in Hem 24 h （P〈0.05）. The injuries of lung tissues after hemorrhagic shock have been demonstrated by histological examination with electron microscope and light microscope. Conclusions： TLR-4 and HO-1 might modulate the bal- ance of pro- and anti-inflammatory processes in inflamma- tory reaction of hemorrhagic shock-induced ALl, and the activation of Toll-like receptor might induce the transcrip- tion activity of HO- 1, which may play a key role in acute lung injury.

Heme oxygenase-1/carbon monoxide signaling participates in the accumulation of triterpenoids of Ganoderma lucidum

Ganoderic triterpenoids(GTs)are the primary bioactive constituents of the Basidiomycotina fungus,Ganoderma lucidum.These compounds exhibit antitumor,anti-hyperlipidemic,and immune-modulatory pharmacological activities.This study focused on GT accumulation in mycelia of G.lucidum mediated by the heme oxygenase-1(HO-1)/carbon monoxide(CO)signaling.Compared with the control,hemin(10μmol/L)induced an increase of 60.1%in GT content and 57.1%in HO-1 activity.Moreover,carbon monoxide-releasing molecule-2(CORM-2),CO donor,increased GT content by 56.0%and HO-1 activity by 18.1%.Zn protoporphyrin IX(ZnPPIX),a specific HO-1 inhibitor,significantly reduced GT content by 26.0%and HO-1 activity by 15.8%,while hemin supplementation reversed these effects.Transcriptome sequencing showed that HO-1/CO could function directly as a regulator involved in promoting GT accumulation by regulating gene expression in the mevalonate pathway,and modulating the reactive oxygen species(ROS)and Ca2+pathways.The results of this study may help enhance large-scale GT production and support further exploration of GT metabolic networks and relevant signaling cross-talk.

Effect of biologically active fraction of Nardostachys jatamansi on cerulein-induced acute pancreatitis

AIM： To determine if the fraction of Nardostachysjata- mansi （N J） has the potential to ameliorate the severity of acute pancreatitis （AP）. METHODS： Mice were administered the biologically active fraction of N J, i.e., the 4th fraction （N J4）, intra- peritoneally, and then injected with the stable chole- cystokinin analogue cerulein hourly for 6 h. Six hours after the last cerulein injection, the pancreas, lung, and blood were harvested for morphological examination,measurement of cytokine expression, and examination of neutrophil infiltration. RESULTS： N J4 administration attenuated the sever- ity of AP and lung injury associated with AP. It also reduced cytokine production and neutrophil infiltration and resulted in the in vivo up-regulation of heine oxy- genase-1 （HO-1）. Furthermore, NJ4 and its biologically active fraction, N J4-2 inhibited the cerulein-induced death of acinar cells by inducing HO-1 in isolated pan- creatic acinar cells. CONCLUSION： These results suggest that N J4 may be a candidate fraction offering protection in AP and N J4 might ameliorate the severity of pancreatitis by induc- ing HO-1 expression.

Protective effects of curcumin in APPswe transfected SH-SY5Y cells

The APPswe plasmid was transfected into the neuroblastoma cell line SH-SY5Y to establish a cell model of Alzheimer＇s disease. Graded concentration and time course experiments demonstrate that curcumin significantly upregulates phosphatidylinositol 3-kinase （PI3K）, Akt, nuclear factor E2-related factor-2 （Nrf2）, heme oxygenase 1 and ferritin expression, and that it significantly downregulates heme oxygenase 2, reactive oxygen species and amyloid-beta 40/42 expression. These effects of curcumin on PI3K, Akt and Nrf2 were blocked by LY294002 （PI3k inhibitor） and NF-E2-related factor-2 siRNA. The results indicate that the cytoprotection conferred by curcumin on APPswe transfected SH-SY5Y cells is mediated by its ability to regulate the balance between heme oxygenase 1 and 2 via the PI3K/Akt/Nrf2 intracellular signaling pathway.

Zinc-substituted hemoglobin with specific drug binding sites and fatty acid resistance ability for enhanced photodynamic therapy

Precisely designed protein-based nanodrugs, as a kind of colloidal drug system, have attracted significant attention in tumor therapy because of their refined drug loading ratio, controlled delivery efficacy and natural biocompatibility. However, most drugs are conjugated to the protein carriers randomly without specific binding sites. Moreover, such sites could easily be replaced by lipophilic molecules in the physiological environment and result in low delivery efficiency. With strong and specific binding locations especially comparatively narrow spatial binding sites and nonflexible structure, hemin (FePPIX)-free hemoglobin or apohemoglobin (apoHb), as a natural metalloporphyrin protein carrier, represents great potential in bioapplication. Therefore, we herein introduce a folate acid (FA) modified, zinc-substituted hemoglobin (ZnPHb-FA) as a naturally occurring protein matrix-based photosensitizer for cancer photodynamic therapy (PDT). Noncovalent inserted ZnPPIX molecules in apoHb possess an extremely stable property and significant recovered photoproperties with superior biocompatibility and phototoxicity, both in vitro and in vivo. This stability was verified by molecular docking analysis and calculation of binding constant, representing a total of five drug binding sites of apoHb for ZnPPIX molecules, four of which are energetically favorable (△G value of -11.9 kcal/mol), and one which is energetically acceptable (△G value of -9 kcal/mol). Folate acid modification has been shown to efficiently enhance the internalization and retention time of ZnPHb nanodrug. ZnPHb-FA is also an efficient depressor of hemin oxygenase-1 (HO-1), which could, in turn, lower the antioxidant ability of cancer cells by decreasing the production of biiirublin. Results in vitro and in vivo both indicated that the firmly combination of apoHb and ZnPPIX described here represents a novel and efficient protein nanodrug systems for cancer therapy.