BLZ945通过诱导巨噬细胞极化抑制恶性胶质瘤生长*

目的：探讨巨噬细胞集落刺激因子1 受体（CSF-1R）抑制剂BLZ945 对恶性胶质瘤生长的作用及其相关机 制。方法：30 只雄性5 ～ 6 周龄Wistar 大鼠随机分为恶性胶质瘤动物模型组（ 模型组）、BLZ945 低剂量治疗组（ 低 剂量组）和BLZ945 高剂量治疗组（ 高剂量组）,使用C6 胶质瘤细胞构建恶性胶质瘤动物模型。观察并记录动物 的生存率;治疗结束后处死大鼠,开颅取全脑,称取脑组织重量,测量并计算肿瘤体积;流式细胞术检测巨噬细 胞免疫表型;逆转录- 聚合酶链反应（RT-PCR）检测γ 干扰素（IFN-γ）和细胞因子信号转导抑制因子3（SOCS3） mRNA 的转录水平;酶联免疫吸附测定（ELISA）检测IFN-γ 和SOCS3蛋白表达水平。结果：BLZ945 可明显抑 制恶性胶质瘤的生长,并显著提高荷瘤鼠的生存率,诱导肿瘤微环境中的巨噬细胞向M1 型极化,上调肿瘤组织 IFN-γ 的表达,并下调SOCS3的表达。结论：BLZ945 通过调节IFN-γ 和SOCS3的表达,诱导巨噬细胞由M2 型 向M1 型极化,激活抗肿瘤免疫,进而抑制恶性胶质瘤的生长。     

Adrenergic regulation of the innate immune response in common carp (Cyprinus carpio L.)

Catecholamines exert their physiological actions through α and β adrenergic receptors (ARs). As ARs are not exclusively expressed on neuroendocrine cells, but also on leukocytes, they may facilitate neuroendocrine modulation of immune responses. We sequenced the β_2a-AR in common carp, and studied its expression profile and involvement in the regulation of teleost innate immune responses.β_2a-AR messenger RNA was found to be constitutively expressed in brain areas, especially in the preoptic nucleus (NPO, homologous to the mammalian hypothalamus), and in immune organs. During the active phase of an in vivo inflammatory response, induced by i.p. zymosan treatment, β_2a-AR gene expression was up-regulated in the peritoneal leukocytes. Additionally, adrenaline in vitro reduced the synthesis of oxygen radical species and nitric oxide, while it enhanced arginase activity in fish phagocytes. Furthermore, in vitro adrenaline administration inhibited expression of pro-inflammatory cytokines, chemokines and their receptors. It is therefore hypothesized that adrenaline will down-regulate phagocyte skewing toward classical/innate polarization.     

七氟醚可抑制氧化型低密度脂蛋白诱导巨噬细胞M1型极化

目的 探究七氟醚通过JAK3/STAT5信号通路对氧化型低密度脂蛋白(oxidized low-density lipoprotein,Ox-LDL)诱导巨噬细胞M1型极化的影响.方法 酶联免疫法检测细胞培养液上清中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(inte...     

The CaSR/TRPV4 coupling mediates pro-inflammatory macrophage function

Aim

Although calcium-sensing receptor (CaSR) and transient receptor potential vanilloid 4 (TRPV4) channels are functionally expressed on macrophages, it is unclear if they work coordinately to mediate macrophage function. The present study investigates whether CaSR couples to TRPV4 channels and mediates macrophage polarization via Ca²⁺ signaling.

Methods

The role of CaSR/TRPV4/Ca²⁺ signaling was assessed in lipopolysaccharide (LPS)-treated peritoneal macrophages (PMs) from wild-type (WT) and TRPV4 knockout (TRPV4 KO) mice. The expression and function of CaSR and TRPV4 in PMs were analyzed by immunofluorescence and digital Ca²⁺ imaging. The correlation factors of M1 polarization, CCR7, IL-1β, and TNFα were detected using q-PCR, western blot, and ELISA.

Results

We found that PMs expressed CaSR and TRPV4, and CaSR activation-induced marked Ca²⁺ signaling predominately through extracellular Ca²⁺ entry, which was inhibited by selective pharmacological blockers of CaSR and TRPV4 channels. The CaSR activation-induced Ca²⁺ signaling was significantly attenuated in PMs from TRPV4 KO mice compared to those from WT mice. Moreover, the CaSR activation-induced Ca²⁺ entry via TRPV4 channels was inhibited by blocking phospholipases A2 (PLA2)/cytochromeP450 (CYP450) and phospholipase C (PLC)/Protein kinase C (PKC) pathways. Finally, CaSR activation promoted the expression and release of M1-associated cytokines IL-1β and TNFɑ, which were attenuated in PMs from TRPV4 KO mice.

Conclusion

We reveal a novel coupling of the CaSR and TRPV4 channels via PLA2/CYP450 and PLC/PKC pathways, promoting a Ca²⁺-dependent M1 macrophage polarization. Modulation of this coupling and downstream pathways may become a potential strategy for the prevention/treatment of immune-related disease.     

巨噬细胞的骨免疫学效应

背景：巨噬细胞以其显著的骨免疫学效应得到学者们的广泛关注,其功能和应用已成为研究热点。目前研究主要涉及巨噬细胞的起源、极化、骨免疫学效应及其在骨修复中的应用。目的：综述巨噬细胞的骨免疫学效应及其在骨修复中应用的研究进展,证实巨噬细胞在骨组织工程中具有卓越的研究价值和应用前景。方法：利用PubMed、Web of Science和CNKI数据库检索2010-2022年发表的相关文献,检索词为“巨噬细胞极化、骨、成骨、骨免疫学、生物材料、组织工程”“macrophage polarization,bone,osteogenesis,osteoimmunology,biomaterials,tissue engineering”,并纳入少量年份久远的经典文献。通过阅读标题和摘要进行初筛,排除与文章主题不相关的文献,最终纳入120篇文献进行综述分析。结果与结论：(1)巨噬细胞包括单核细胞来源的炎性巨噬细胞和组织驻留巨噬细胞,其中不同组织的驻留巨噬细胞具有不同的发育起源组合,绝大多数组织驻留巨噬细胞起源于胚胎时期的卵黄囊;(2)巨噬细胞具有高度可塑性,在不同刺激下极化为M1/M2表型,分别释放促...     

小胶质细胞极化介导炎症反应在脊髓损伤中的作用

背景:小胶质细胞极化参与脊髓损伤后的炎症反应,并在其中发挥关键作用。相关研究表明,有效诱导小胶质细胞从M1促炎表型向M2抗炎表型极化,可以减轻脊髓损伤后的炎症反应,促进组织的修复再生和神经功能的恢复。目的:文章对小胶质细胞的功能和极化、小胶质细胞极化对脊髓损伤的影响及其潜在调控策略以及脊髓损伤后炎症反应进行综述。方法:检索PubMed、Web of Science和中国知网数据库,英文检索词为“microglia,polarization,spinal cord injury,inflammation”,中文检索词为“小胶质细胞、极化、脊髓损伤、炎症”,按纳入和排除标准共纳入80篇文献进行总结。结果与结论:①由小胶质细胞介导的稳定而持续的炎症反应,对脊髓损伤的预后至关重要。②在生理条件下,小胶质细胞处于M0静止表型,但在脊髓损伤后,小胶质细胞活化,进而极化成M1促炎表型,导致神经组织修复能力降低和出现持续性神经炎症。③在脊髓损伤的炎症反应过程中,调控小胶质细胞向M2表型极化或至少向M2表型倾斜,有利于抑制氧化应激反应、调节突触重塑、促进轴突再生和血管生成,是一种有效的调控策略。④截止到目前的研究表明,间充质干细胞、外泌体、临床药物、天然产物、miRNAs和靶点分子可调控小胶质细胞在M1和M2表型之间的转换,这为脊髓损伤后神经组织的修复提供了一种新的思路,未来需进一步研究小胶质细胞在脊髓损伤过程中调控极化的详细机制。     

小胶质细胞M1型极化诱发小鼠焦虑样行为及其对前额叶神经元树突棘密度的影响

目的:焦虑样行为与脑内小胶质细胞M1型极化关系密切,但M1型小胶质细胞如何作用于神经元导致焦虑样行为并不清楚,因此本研究的目的在于观察小鼠脑内小胶质细胞向M1型极化诱发产生焦虑样行为,并探讨其对神经元树突棘的影响。方法:实验组采用脂多糖(lipopolysaccharide,LPS)脑内注射诱导C57小鼠小胶质细胞向M1型极化,通过旷场实验和高架十字迷宫实验观察小鼠的行为学改变;应用免疫荧光染色方法观察实验组和对照组小鼠的内侧前额叶皮质(medial prefrontal cortex,m PFC)内一氧化氮合酶(i NOS)与小胶质细胞标志物iba-1的表达变化,通过Western Blot方法观察实验组和对照组小鼠的m PFC内i NOS、致炎因子白介素1(IL-1)β、肿瘤坏死因子(TNF)-α的表达变化,以i NOS、IL-1β和TNF-α表达量均显著增加作为小胶质细胞向M1型极化的标志。采用高尔基(Golgi)染色观察m PFC内神经元树突棘的变化。结果:(1)实验组小鼠mPFC内小胶质细胞i NOS、IL-1β和TNF-α表达量均升高,提示小胶质细胞向M1型极化。(2)实验组小鼠在旷场内的中央活动距离和中央活动时间比对照组均明显减少(P0.001);实验组小鼠在高架十字迷宫开臂停留时间比正常组明显减少(P0.05)。(3)Golgi染色结果显示实验组小鼠mPFC内神经元树突棘的密度相较于对照组增多(P0.05)。结论:mPFC内小胶质细胞M1型极化可诱发小鼠产生焦虑样行为,并伴随mPFC内神经元树突棘密度增多,提示脑内小胶质细胞M1型极化可诱导焦虑样行为,可能与内侧前额叶皮质锥体神经元树突棘可塑性改变相关。     

诱导共刺激分子转基因小鼠类风湿性关节炎中滤泡辅助性T细胞的极化

目的 探讨可诱导共刺激分子(ICOS)转基因小鼠类风湿性关节炎(RA)模型中ICOS信号对滤泡辅助性T细胞(Tfh)极化的影响.方法 1.流式细胞仪分析测定ICOS转基因(ICOS-Trangenic,ICOS-Tg)小鼠及其野生型对照(WT)小鼠脾细胞CD4+T淋巴细胞共刺激分子ICOS在不同时期的表达趋势及特征;2.ELISA检测脾淋巴细胞培养悬液中干扰素(IFN)-γ、IL-21、IL-23、IL-17的水平;结果1.野生型RA小鼠脾CD4+T淋巴细胞表面的ICOS的表达水平从4w到12w为上升趋势(％)(4 W:6.5 ±1.0;7 W:13.2±1.3;12 W:23.5±:2.1);ICOS-Tg小鼠脾CD4+T淋巴细胞表面ICOS的表达三期同样呈上升趋势(％)(4W:8.2±0.9;7W:17.2±1.5;12W:31.6 ±3.0),但与野生型小鼠相比各期均表达上调(均P＜0.05);2.野生型小鼠IFN-γ从4W开始表达上升,7周达峰值后下降,ICOS-Tg小鼠同野生型小鼠相比,趋势相同但表达在各期呈下调趋势,差异有统计学意义(4 W～20 W均P＜0.05);野生型小鼠的IL-21、IL-23及IL-17的表达于4周上升,12周达峰值后下降,ICOS-Tg小鼠同野生型小鼠相比,三者的各期表达趋势相同但呈上调趋势(IL-21和IL-17有统计学意义,P＜0.05;IL-23无统计学意义,P＞0.05).结论 RA小鼠在其致病过程中共刺激信号ICOS呈上调表达;ICOS-Tg小鼠同野生型小鼠相比,其脾淋巴细胞培养上清中IFN-γ呈显著下调趋势;Tfh分化相关的细胞因子IL-21、IL-17则均显著上调表达.Tfh细胞及其作用因子很可能参与了RA的免疫应答,与RA的发生发展有关.     

IRF1对M1巨噬细胞极化及其抗肿瘤效应的影响

目的研究IRF1对M1巨噬细胞极化及M1介导的抗肝癌细胞增殖和凋亡的影响。方法构建单核细胞U937来源M1巨噬细胞模型(U937-M1),将细胞分为4组:用PMA诱导的未活化巨噬细胞组(M0),用PMA,IFN-γ和LPS处理的M1型巨噬细胞组(M1),用siRNA干扰IRF1的M1型巨噬细胞组(si IRF1)以及阴性干扰的M1型巨噬细胞组(si C)。用流式细胞术检测M1/M2特异表面标志物CD86/CD206的表达;q PCR检测M1/M2相关基因(IL-12p35,IL-12p40,IL-23p19,IL-6,TNF-α/IL-10)及IFNB1的表达;ELISA检测IL-12p70,IL-10及IFN-β的表达;Western blot检测IRF1及IRF5的表达;CCK8和流式细胞术分别检测Hep G2及SMMC-7721增殖和凋亡。结果与U937-M1组相比,干扰IRF的M1组CD86表达降低,但CD206升高(P0.05);mRNA水平上,IL-12p35,IL-12p40,IL-23p19,IL-6,TNF-α以及IFNB1表达降低,但IL-10表达增高(P0.01);蛋白水平上,IL-12p70,IFN-β及IRF5表达降低,但IL-10表达增高(P0.05)。IRF1干扰后,M1巨噬细胞促进肝癌细胞增殖、抑制其凋亡(P0.05)。结论干扰IRF1后,M1巨噬细胞极化状态受损,甚至部分向M2型转变;其抗肿瘤效应转变为促肿瘤效应;且IRF1可能参与调节IFN-β与IRF5的表达。