Toxoplasma ROP16Ⅰ/Ⅲ ameliorated inflammatory bowel diseases via inducing M2 phenotype of macrophages

BACKGROUND Inflammatory bowel disease（IBD） is characterized by chronic and non-specific inflammation of the intestinal mucosa and mainly includes ulcerative colitis and Crohn’s disease.AIM To explore the beneficial effect of Toxo ROP16Ⅰ/Ⅲ-induced M2 phynotype macrophages in homeostasis of IBDs through downregulation of M1 inflammatory cells.METHODS RAW264.7 macrophages stimulated by lipopolysaccharide（LPS）（M1 cells） were co-cultured with Caco-2 cells as an inflammatory model of IBD in vitro.The expression of Toxo ROP16Ⅰ/Ⅲ was observed in RAW264.7 macrophages that were transfected with p EGFP-rop16Ⅰ/Ⅲ.The phenotypes of M2 and M1 macrophage cells were assessed by quantitative real-time reverse transcriptase polymerase chain reaction and the expression of tumor necrosis factor（TNF）-α,interleukin（IL）-1β,IL-6,transforming growth factor（TGF）-β1,IL-10,inducible nitric oxide synthase（i NOS）,and arginase-1（Arg-1） was detected.The expression of i NOS,Arg-1,signal transducer and activator of transcription 3（Stat3）,p-Stat3,Stat6,pStat6,programmed death ligand-2（PD-L2）,caspase-3,-8,and-9 was analyzed by Western blotting,and Griess assays were performed to detect nitric oxide（NO）.TNF-α,IL-1β,IL-6,TGF-β1,and IL-10 expression in the supernatants was detected by enzyme-linked immunosorbent assay,and Caco-2 cell apoptosis was determined by flow cytometry after mixing M1 cells with M2 cells in a Caco-2 cell co-culture system.RESULTS M1 cells exhibited significantly increased production of i NOS,NO,TNF-α,IL-1β,and IL-6,while Toxo ROP16Ⅰ/Ⅲ induced macrophage bias to M2 cells in vitro,showing increased expression of Arg-1,IL-10 and TGF-β1 and elevated production of p-Stat3 and p-Stat6.The mixed M1 and M2 cell culture induced by Toxo ROP16 Ⅰ/Ⅲ exhibited decreased production of NO and i NOS and upregulated expression of Arg-1 and PD-L2.Accordingly,Caco-2 cells became apoptotic,and apoptosis-associated proteins such as caspase-3,-8 and-9 were dampened during co-culture of M1 and M2 cells.Flow cytometry analysis showed that co-culture of M1 cells with Caco-2 cells facilitated the apoptosis of Caco-2 cells,but co-culture of M1 and M2 cells alleviated Caco-2 cell apoptosis.CONCLUSION Toxo ROP16 Ⅰ/Ⅲ-induced M2 macrophages inhibited apoptosis of Caco-2 cells caused by M1 macrophages.This finding may help gain a better understanding of the underlying mechanism and represent a promising therapeutic strategy for IBDs.

Microglia:a promising therapeutic target in spinal cord injury

Microglia are present throughout the central nervous system and are vital in neural repair,nutrition,phagocytosis,immunological regulation,and maintaining neuronal function.In a healthy spinal cord,microglia are accountable for immune surveillance,however,when a spinal cord injury occurs,the microenvironment drastically changes,leading to glial scars and failed axonal regeneration.In this context,microglia vary their gene and protein expression during activation,and proliferation in reaction to the injury,influencing injury responses both favorably and unfavorably.A dynamic and multifaceted injury response is mediated by microglia,which interact directly with neurons,astrocytes,oligodendrocytes,and neural stem/progenitor cells.Despite a clear understanding of their essential nature and origin,the mechanisms of action and new functions of microglia in spinal cord injury require extensive research.This review summarizes current studies on microglial genesis,physiological function,and pathological state,highlights their crucial roles in spinal cord injury,and proposes microglia as a therapeutic target.

Function of microglia and macrophages in secondary damage after spinal cord injury

Spinal cord injury （SCI） is a devastating type of neurological trauma with limited therapeutic op- portunities. The pathophysiology of SCI involves primary and secondary mechanisms of injury. Among all the secondary injury mechanisms, the inflammatory response is the major contrib- utor and results in expansion of the lesion and further loss of neurologic function. Meanwhile, the inflammation directly and indirectly dominates the outcomes of SCI, including not only pain and motor dysfunction, but also preventingneuronal regeneration. Microglia and macrophages play very important roles in secondary injury. Microglia reside in spinal parenchyma and survey the microenvironment through the signals of injury or infection. Macrophages are derived from monocytes recruited to injured sites from the peripheral circulation. Activated resident microglia and monocyte-derived macrophages induce and magnify immune and inflammatory responses not only by means of their secretory moleculesand phagocytosis, but also through their influence on astrocytes, oligodendrocytes and demyelination. In this review, we focus on the roles of mi- croglia and macrophages in secondary injury and how they contribute to the sequelae of SCI.

水母雪莲细胞悬浮培养合成黄酮及抗氧化活性(英文)

对水母雪莲细胞悬浮培养过程中细胞生长、黄酮积累和底物消耗的动力学过程进行了研究。经15d液体培养可获得最大生物量干重和黄酮产量分别为17.2g·L-1和607.8mg·L-1,通过调控基本培养基种类和有机添加物可提高雪莲细胞的生长和黄酮积累。获得的水母雪莲细胞培养物具有明显的抗氧化活性,其抗氧化活性与雪莲细胞中的黄酮含量相关。

TET2上调IRAK-M表达促进巨噬细胞内毒素耐受

目的:研究TET2蛋白(ten-eleven translocation 2)对巨噬细胞白介素-1受体相关激酶-M(interleukin-1 receptor-associated kinase-M,IRAK-M)表达的影响,探讨细菌内毒素(endotoxin/lipopolysaccharide,LPS)刺激诱导TET2表达的上游信号通路。方法:用LPS刺激小鼠巨噬细胞系RAW264.7诱导内毒素耐受,qPCR和Western blot检测LPS刺激不同时间点和内毒素耐受时IRAK-M和TET2表达;用siRNA干扰TET2表达,qPCR和Western blot检测对IRAK-M表达的影响;用促分裂原活化的蛋白激酶(mitogen-activated protein kinase,MAPK)信号通路抑制剂预处理细胞,再用LPS刺激,qPCR和Western blot检测对TET2表达的影响。结果:TET2蛋白和IRAK-M在LPS刺激RAW264.7细胞诱导内毒素耐受后仍持续表达,且表达模式都和细胞因子表达模式相反;干扰TET2表达抑制IRAK-M m RNA(P=0.000)和蛋白水平表达;抑制MAPK信号通路抑制TET2表达。结论:巨噬细胞炎症反应过程中MAPK信号通路上调TET2表达,TET2通过促进IRAK-M表达促进内毒素耐受。

巨噬细胞极化在2型糖尿病和心血管疾病中的作用

巨噬细胞具有多种亚型。经典激活的巨噬细胞(M1)和替代激活的巨噬细胞(M2)是巨噬细胞不均一性的两种极端分类,分别由Th1和Th2细胞因子激活。M1和M2巨噬细胞的表面标志物、产生的细胞因子和趋化因子及其功能都有明显不同。巨噬细胞的极化由多种分子调控,转录因子是其中最重要的群体之一。极化的巨噬细胞存在于不同的微环境中,扮演着不同角色。身体的代谢状态在很大程度上取决于极化的巨噬细胞和脂肪细胞、肝细胞、骨骼肌细胞之间的相互作用。这些相互作用对2型糖尿病的发生发展起重要作用。极化的巨噬细胞和血管平滑肌细胞、内皮细胞、心肌细胞之间的相互作用很可能会影响心血管疾病的发生发展。这些相互作用导致抗炎的M2巨噬细胞通常在2型糖尿病和心血管疾病中发挥保护作用。这些研究进一步拓展了对生理和病理上巨噬细胞极化功能的认识,并提示了众多临床治疗设想。

The transcription factor PU.1 promotes alternative macrophage polarization and asthmatic airway inflammation

The transcription factor PU.1 is involved in regulation of macrophage differentiation and maturation.However,the role of PU.1 in alternatively activated macrophage(AAM)and asthmatic inflammation has yet been investigated.Here we report that PU.1 serves as a critical regulator of AAM polarization and promotes the pathological progress of asthmatic airway inflammation.In response to the challenge of DRA(dust mite,ragweed,and Aspergillus)allergens,conditional PU.1-deficient(PU/ER(T)^(+/-))mice displayed attenuated allergic airway inflammation,including decreased alveolar eosinophil infiltration and reduced production of IgE,which were associated with decreased mucous glands and goblet cell hyperplasia.The reduced asthmatic inflammation in PU/ER(T)^(+/-) mice was restored by adoptive transfer of IL-4-induced wild-type(WT)macrophages.Moreover,after treating PU/ER(T)^(+/-) mice with tamoxifen to rescue PU.1 function,the allergic asthmatic inflammation was significantly restored.In vitro studies demonstrate that treatment of PU.1-deficient macrophages with IL-4 attenuated the expression of chitinase 3-like 3(Ym-1)and resistin-like molecule alpha 1(Fizz-1),two specific markers of AAM polarization.In addition,PU.1 expression in macrophages was inducible in response to IL-4 challenge,whichwas associated with phosphorylation of signal transducer and activator of transcription 6(STAT6).Furthermore,DRAchallenge in sensitized mice almost abrogated gene expression of Ym-1 and Fizz-1 in lung tissues of PU/ER(T)^(+/-) mice compared with WT mice.These data,all together,indicate that PU.1 plays a critical role in AAM polarization and asthmatic inflammation.

靶向干预巨噬细胞极化的矽肺治疗药物

矽肺是我国职业病中最常见、危害最严重的一类疾病,其发病机制与肺巨噬细胞密切相关。二氧化硅(SiO_(2))进入肺内被巨噬细胞吞噬后使巨噬细胞发生极化,极化后的巨噬细胞分为经典活化型巨噬细胞(M1)和选择活化型巨噬细胞(M2),它们分别通过复杂的调控机制释放各种炎性因子,最终导致肺纤维化。本文拟就巨噬细胞极化与矽肺纤维化的关系,以及靶向干预巨噬细胞极化的矽肺治疗药物进行综述,以期为深入探索矽肺纤维化的发病机制和寻找新的治疗方法提供理论依据。