Anti-Oxidant,Anti-Hemolytic Effects of Crataegus aronia Leaves and Its Anti- Proliferative Effect Enhance Cisplatin Cytotoxicity in A549 Human Lung Cancer Cell Line

Objective: For Arabian traditional medicine, Crataegus aronia syn. Azarolus (L) Bosc. ex DC (Rosaceae) is widely used to treat diabetes, sexual weakness, cardiovascular diseases and cancer. The anti-cancerous and anti-hemolysis effects of the hydroalcoholic extract of this plant have never been investigated before. The present study aims to evaluate the biological activities of the hydroalcoholic extract of Crataegus aronia leaves in combination with cisplatin, one of the most widely employed chemotherapeutics, on A549 human lung cancer cell line. Methods: The anti-oxidant and anti-proliferative activities of leaves, fruits, seeds of C. aronia were investigated by DPPH method and MTT assay; respectively. Cell migration activity was investigated by wound healing and by cell aggregation assays. The effect of C. aronia in inducing cell cycle arrest along with activating cell apoptosis was evaluated by flow cytometry and Western blot assays, respectively. Results: Our results showed that C. aronia leaves (C. aronia L.) had the highest anti-oxidant and anti-proliferative activities. The leaves extract was potent against hemolysis of the human erythrocytes and showed elevated decrease in migration by reducing wound healing migration and by increasing cell aggregation. Finally, C. aronia L. treatment exhibited apoptotic activity on A549 cells by the down-regulation of PARP-1, caspase-3 and Bcl-2 proteins and by increasing the percentage of A549 cells in sub G0 cell cycle. Moreover, the co-treatment of C. aronia L. and cisplatin remarkably sensitised A549 cells to cisplatin. Conclusion: The results suggested that C. aronia L. could be used as a potential treatment against human lung cancer exhibiting minimal side effects on human health.     

Interstitial lung diseases in children

Interstitial lung disease (ILD) in children (chILD) is a heterogeneous group of rare respiratory disorders that are mostly chronic and associated with high morbidity and mortality. The pathogenesis of the various chILD is complex and the diseases share common features of inflammatory and fibrotic changes of the lung parenchyma that impair gas exchanges. The etiologies of chILD are numerous. In this review, we chose to classify them as ILD related to exposure/environment insults, ILD related to systemic and immunological diseases, ILD related to primary lung parenchyma dysfunctions and ILD specific to infancy. A growing part of the etiologic spectrum of chILD is being attributed to molecular defects. Currently, the main genetic mutations associated with chILD are identified in the surfactant genes SFTPA1, SFTPA2, SFTPB, SFTPC, ABCA3 and NKX2-1. Other genetic contributors include mutations in MARS, CSF2RA and CSF2RB in pulmonary alveolar proteinosis, and mutations in TMEM173 and COPA in specific auto-inflammatory forms of chILD. However, only few genotype-phenotype correlations could be identified so far. Herein, information is provided about the clinical presentation and the diagnosis approach of chILD. Despite improvements in patient management, the therapeutic strategies are still relying mostly on corticosteroids although specific therapies are emerging. Larger longitudinal cohorts of patients are being gathered through ongoing international collaborations to improve disease knowledge and targeted therapies. Thus, it is expected that children with ILD will be able to reach the adulthood transition in a better condition.     

多聚嘧啶序列结合蛋白相关剪接因子对缺氧诱导人视网膜微血管内皮细胞功能的影响

目的观察多聚嘧啶序列结合蛋白相关剪接因子(PSF)对缺氧诱导的人视网膜微血管内皮细胞(hRMECs)功能的影响。方法采用三质粒系统构建慢病毒颗粒(LV)-PSF。LV-PSF体外感染hRMECs后通过流式细胞计数法测定其感染效率。应用实时定量PCR(RT-PCR)检测经LV-PSF感染的hRMECs中PSF mRNA表达水平。将实验分为体内和体外两部分。体内实验:7日龄健康C57B/L6小鼠20只,采用随机数字表法分为正常组、氧诱导视网膜病变(OIR)组、OIR+LV-空载体(Vec)组和OIR+LV-PSF组,每组5只。除正常组外,其余3组构建OIR模型。OIR组除缺氧刺激外,不做其余处理。OIR+LV-Vec组和OIR+LV-PSF组小鼠分别玻璃体腔注射LV-Vec或LV-PSF。观察LV-PSF对视网膜新生血管(RNV)形成的影响。体外实验:将hRMECs分为正常组、缺氧组、空载组、PSF高表达组。正常组为正常体外培养的hRMECs;缺氧组为缺氧刺激3 h恢复正常培养条件24 h的hRMECs;空载组、PSF高表达组分别为用LV-Vec、LV-PSF感染48 h,再用缺氧刺激3 h恢复正常培养条件24 h的hRMECs。采用MTT比色法观察PSF对细胞增生能力的影响。采用细胞划痕实验和Transwell迁移实验观察PSF对缺氧刺激下细胞迁移能力的影响。采用RT-PCR观察各组细胞中HIF-1α、VEGF及PSF的mRNA表达。结果成功构建可稳定高表达PSF的LV-PSF,流式细胞仪测定其感染效率为97%,RT-PCR测得经LV-PSF感染的hRMECs中PSF mRNA水平明显上调。体内实验:OIR组、OIR+LV-Vec组小鼠RNV面积较正常组明显增加(t=18.31、43.71),OIR+LV-PSF组小鼠RNV面积较OIR组(t=11.30)、OIR+LV-Vec组(t=15.47)明显减小,差异均有统计学意义(P<0.05)。体外实验:MTT比色法检测结果显示,缺氧组hRMECs增生能力较正常组明显增强(t=2.57),PSF高表达组hRMECs增生能力较正常组、缺氧组、空载组明显降低(t=5.26、5.46、3.73),差异均有统计学意义(P<0.05)。细胞划痕实验结果显示,缺氧刺激3 h恢复正常条件24 h或48 h均可刺激缺氧组与空载组细胞明显迁移(t=8.35、13.84,P<0.05);而与缺氧组与空载组比较,PSF高表达组细胞迁移不明显(t=10.99、18.27、9.75、8.93、26.94、7.01,P<0.05)。Transwell小室实验结果显示,正常组和空载组微孔膜上染色的细胞数较多,而PSF高表达组微孔膜上染色的细胞数明显减少(t=9.33、6.15,P<0.05)。RT-PCR检测结果显示,与正常组比较,缺氧组、空载组hRMECs中HIF-1α、VEGF的mRNA表达明显增加(t=15.23、21.09,P<0.05),PSF mRNA表达无明显变化(t=0.12、2.15,P<0.05);与缺氧组、空载组比较,PSF高表达组hRMECs中HIF-1α、VEGF的mRNA表达明显下降(t=10.18、13.10,P<0.05),PSF mRNA表达明显增加(t=65.00、85.79,P<0.05)。结论PSF可减少OIR模型小鼠RNV面积。PSF可能通过HIF-1α/VEGF信号通路抑制缺氧诱导的hRMECs增生和迁移。     

Effects of Long-time Exposure to Lipopolysaccharide on Intestinal Lymph Node Immune Cells and Antibodies Level in Mice

Background: Endotoxin, widely present in the living environment of humans and animals, leads to endotoxemia during a short period. However, the long-term effects of endotoxin on immune function are unclear. Objective: To determine the importance of long-term endotoxin treatment on function of immune system. Methods: The mice were treated with different doses of lipopolysaccharide (LPS) for a month; the collected samples were then analyzed in terms of value changes in hematological parameters, lymphocyte subtypes, and immunoglobulins level. Results: The number of monocytes (MONO) and neutrophils (NEU) in the three treatment groups was significantly lower than the control after 30 days. However, the proportion of CD8+ T lymphocytes showed a rising trend in the mesenteric lymph nodes (MLNs) and Peyer's patches (PPs) while the CD4+ T cell was reduced. At the same time, a decrease was observed in the percentage of CD19+CD38+ B lymphocytes. Interestingly, the change of lymphocytes in PPs was more significant than that in MLNs, suggesting that immune response in the PPs occurred before the MLNs. Consistent with the changes in B cells, the content of IgA and IgG showed a downward trend. Conclusion: Long-term exposure to low-dose endotoxin had little or no effect on the immune function of the body, suggesting that the endotoxin can be rapidly eliminated by the immune system. Nonetheless, the number of immune cells was reduced in the high-dose group. T- and B-lymphocytes were significantly reduced, resulting in a decrease in immunoglobulin level, and showing a significant immune suppression state.     

Prostaglandin D2 synthase modulates macrophage activity and accumulation in injured peripheral nerves

We have previously reported that prostaglandin D2 Synthase (L-PGDS) participates in peripheral nervous system (PNS) myelination during development. We now describe the role of L-PGDS in the resolution of PNS injury, similarly to other members of the prostaglandin synthase family, which are important for Wallerian degeneration (WD) and axonal regeneration. Our analyses show that L-PGDS expression is modulated after injury in both sciatic nerves and dorsal root ganglia neurons, indicating that it might play a role in the WD process. Accordingly, our data reveals that L-PGDS regulates macrophages phagocytic activity through a non-cell autonomous mechanism, allowing myelin debris clearance and favoring axonal regeneration and remyelination. In addition, L-PGDS also appear to control macrophages accumulation in injured nerves, possibly by regulating the blood–nerve barrier permeability and SOX2 expression levels in Schwann cells. Collectively, our results suggest that L-PGDS has multiple functions during nerve regeneration and remyelination. Based on the results of this study, we posit that L-PGDS acts as an anti-inflammatory agent in the late phases of WD, and cooperates in the resolution of the inflammatory response. Thus, pharmacological activation of the L-PGDS pathway might prove beneficial in resolving peripheral nerve injury.     

The fate and function of oligodendrocyte progenitor cells after traumatic spinal cord injury

Oligodendrocyte progenitor cells (OPCs) are the most proliferative and dispersed population of progenitor cells in the adult central nervous system, which allows these cells to rapidly respond to damage. Oligodendrocytes and myelin are lost after traumatic spinal cord injury (SCI), compromising efficient conduction and, potentially, the long-term health of axons. In response, OPCs proliferate and then differentiate into new oligodendrocytes and Schwann cells to remyelinate axons. This culminates in highly efficient remyelination following experimental SCI in which nearly all intact demyelinated axons are remyelinated in rodent models. However, myelin regeneration comprises only one role of OPCs following SCI. OPCs contribute to scar formation after SCI and restrict the regeneration of injured axons. Moreover, OPCs alter their gene expression following demyelination, express cytokines and perpetuate the immune response. Here, we review the functional contribution of myelin regeneration and other recently uncovered roles of OPCs and their progeny to repair following SCI.     

Schwann cells: Rescuers of central demyelination

The presence of peripheral myelinating cells in the central nervous system (CNS) has gained the neurobiologist attention over the years. Despite the confirmed presence of Schwann cells in the CNS in pathological conditions, and the long list of their beneficial effects on central remyelination, the cues that impede or allow Schwann cells to successfully conquer and remyelinate central axons remain partially undiscovered. A better knowledge of these factors stands out as crucial to foresee a rational therapeutic approach for the use of Schwann cells in CNS repair. Here, we review the diverse origins of Schwann cells into the CNS, both peripheral and central, as well as the CNS components that inhibit Schwann survival and migration into the central parenchyma. Namely, we analyze the astrocyte- and the myelin-derived components that restrict Schwann cells into the CNS. Finally, we highlight the unveiled mode of invasion of these peripheral cells through the central environment, using blood vessels as scaffolds to pave their ways toward demyelinated lesions. In short, this review presents the so far uncovered knowledge of this complex CNS-peripheral nervous system (PNS) relationship.