抗孕激素RU486和ZK98734对双池培养的颗粒细胞和内泡膜细胞分泌功能的影响

本文观察了RU486和ZK98734对培养在羊膜双池培养系统中的猪卵巢颗粒细胞和内泡膜细胞在甾体激素生成过程中的影响。生长在羊膜两书侧的颗粒细胞和内泡膜细胞在加入或不加FSH、LH及不同浓度RU486或ZK98734的条件下培养48h。用RIA测定内、外池培养液中的孕酮（P）和雌二醇（E2）的浓度并与单独培养的结果相比较，同时亦与大鼠颗粒细胞的结果作比较。结果表明：1．有FSH刺激时，两种抗孕激素均明显抑制双池培养中颗粒细胞的P和E2产量；2．不论有或无LH刺激，两种抗孕激素均显著抑制双池培养中内泡膜细胞的P产量；3．双池培养系统模拟了两种卵巢细胞在体内的旁分泌调节关系，比单独培养更加合理；4与大鼠相比，猪卵巢细胞体外培养是研究避孕药对卵巢功能影响的适合模型。     

间充质干细胞治疗肝纤维化的研究进展

肝纤维化是肝脏对慢性炎症、坏死或其他损伤的修复反应,肝硬化则是肝纤维化的终末结局.肝纤维化、肝硬化严重影响着人类健康,但其治疗仍是尚未完全攻破的难题.目前,肝移植是比较有效的治疗手段之一.但是,受到肝源缺乏、并发症多、排异反应和价格昂贵等诸多因素的影响,致使其难以广泛开展.在少数接受了肝移植的患者中,3、12、36个月的生存率分别是94％、 88％、79％.因此,寻找一种有效的替代疗法来治疗这种危及生命的疾病具有重要意义.近年来,以干细胞为基础的细胞疗法从动物模型和临床患者身上均观察到良好的效应,现就肝细胞、肝星状细胞(hepatic stellate cells,HSCs)、旁分泌机制在间充质干细胞(mesenchymal stem cells,MSCs)治疗肝纤维化中的作用机制及其临床应用前景等方面进行综述.     

Fetal and perinatal stem cells in cardiac regeneration: Moving forward to the paracrine era

Cardiovascular disease (CD) is a major burden for Western society. Regenerative medicine has provided encouraging results, yet it has not addressed the focal defects causing CD and mainly related to the inefficient repair programme of the heart. In this scenario, stem cells have been broadly investigated and their paracrine effect proposed as a possible working strategy to boost endogenous mechanisms of repair and regeneration from within the cardiac tissue.The scientific community is now focusing on identifying the most effective stem cell secretome, as the whole of bioactive factors and extracellular vesicles secreted by stem cells and endowed with regenerative potential. Indeed, the adult stem cell-paracrine potential for cardiac regeneration have been widely analyzed with positive outcome. Nevertheless, low yield, invasive sampling and controversial self-renewal may limit adult stem cell application. On the contrary, fetal and perinatal stem cells, which can be easily isolated from leftover sample via prenatal screening during gestation or as clinical waste material after birth, can offer an ideal alternative. These broadly multipotent immature progenitors share features with both adult and embryonic stem cells, show high self-renewal, but they are not tumorigenic neither cause any ethical concern. While fetal and perinatal stem cells demonstrated to improve cardiac function when injected in the injured heart, the comprehensive characterization of their secretome for future applications is still at its infancy.In this review, we will discuss the paracrine potential of the fetal and perinatal stem cell secretome to provide cardiac repair and resurge the dormant mechanisms of cardiac regeneration for future therapy.     

Follistatin suppresses FSHbeta but increases LHbeta expression in the goldfish - evidence for an activin-mediated autocrine/paracrine system in fish pituitary

Our previous study demonstrated that recombinant goldfish activin B stimulated goldfish FSHbeta but inhibited LHbeta expression. Similar to activin B, activin A also exhibited the inverse effects on the expression of the two gonadotropins. The novel dual effects of activins on FSH and LH in the goldfish raise an interesting question as to where the activin comes from in vivo. In the present study, we first demonstrated the expression of activin, its receptors and binding protein follistatin in the goldfish pituitary, leading to a suggestion that an autocrine/paracrine regulatory system involving activin is operative in fish pituitary. To investigate the functionality of the pituitary-derived activin system in the regulation of gonadotropin biosynthesis, we further examined the effects of follistatin, an activin-binding protein, on goldfish FSHbeta and LHbeta expression. Follistatin not only reversed the effects of exogenous activin on FSHbeta and LHbeta expression but also had inverse effects on the basal expression of the genes; and its effects were opposite to those of activin. This suggests that the endogenous activin plays roles in controlling the expression of both FSHbeta and LHbeta genes. It is conceivable that any factors that influence the intrapituitary activin system in vivo will likely affect the biosynthesis of the two gonadotropins in the goldfish.     

基于网络药理学探讨趋化因子-13对间充质干细胞增殖和迁移的影响

目的 以网络药理学技术探讨趋化因子-13（CXCL-13）对人骨髓间充质干细胞（BMSCs）增殖和迁移的影响。 方法 在线数据库预测CXCL-13作用于BMSCs的靶点。Metascape数据库对靶点的基因本体论和京都基因与基因组百科全书(KEGG）信号通路进行富集分析。STRING 11.0数据库进行蛋白质相互作用分析,Cytoscape 3.8的cytoHubba 0.1插件筛选核心基因编码的蛋白质。BMSCs分为对照组、CXCL-13组和PI3K抑制剂组。分别以MTT、流式细胞术和Transwell细胞小室迁移实验检测各组BMSCs的吸光度（A）值、细胞凋亡率和细胞迁移数目情况;ELISA检测各组BMSCs上清液表皮生长因子(EGF)和血管内皮生长因子(VEGF)蛋白含量。Western blotting检测各组BMSCs的Akt、磷酸化Akt（p-Akt）蛋白的表达。 结果 CXCL-13作用于BMSCs 21个靶点。与细胞增殖相关的生物学过程包括干细胞增殖、调节内皮细胞增殖、正向调控平滑肌细胞增殖等32条;与细胞迁移相关的生物学过程包括调节细胞迁移、阿米巴状细胞迁移、调节内皮细胞迁移等22条。KEGG通路包括癌症途径、PI3K-Akt信号通路、MAPK信号通路等40条。核心蛋白包括肿瘤蛋白P53（TP53）、表皮生长因子受体（EGFR）、90kD热休克蛋白αB1（HSP90AB1）、蛋白激酶Cα（PRKCA）、雌激素受体2（ESR2）及前列腺素E受体4（PTGER4）。与其他组相比,CXCL-13组BMSCs的吸光度（A）值和细胞迁移数目均显著增高（P＜0.01,n=15）,细胞凋亡率明显降低（P＜0.01,n=15）;PI3K抑制剂组BMSCs的A值、细胞凋亡率和细胞迁移数目与CXCL-13组相比均呈相反变化（P＜0.01,n=15）。相对于对照组,CXCL-13组BMSCs的EGF和VEGF蛋白含量显著提高（P＜0.01,n=15）,Akt和p-Akt相对表达均明显升高（P＜0.01,n=9）;而PI3K抑制剂组EGF和VEGF蛋白含量、Akt和p-Akt相对表达呈相反变化。 结论 CXCL-13激活PI3K-Akt通路促进BMSCs旁分泌EGF和VEGF蛋白,提高BMSCs增殖和迁移,抑制BMSCs凋亡。     

Functional melanocortin-2 receptors are expressed by mouse aorta-derived mesenchymal progenitor cells

A local melanocortin system is active during tissue injury and inflammation. Thus far this system has been described as autocrine in nature where local production of pro-opiomelanocortin (POMC) peptides by leukocytes feeds back on melanocortin receptor (MC-R) expressing immune cells to quell inflammatory cytokine production. Here we present evidence that POMC peptides may generate extracellular matrix (ECM) changes by inducing matrix production by cells of the mesenchymal lineage through activation of the MC2-R. Using immunoblot, we determined that mouse aorta-derived mesenchymal progenitor cells express both MC2-R and MC3-R. These progenitors respond to treatment with ACTH by increasing collagen matrix synthesis as assessed by picrosirius red stain and (3)H-proline incorporation. ACTH also induces transient increases in intracellular calcium ([Ca(2+)](i)) as assessed using the fluorescent Ca(2+) indicator, fura-2. The ACTH-induced changes in [Ca(2+)](i) are consistent with MC2-R signaling and consist of both an intracellular release and an extracellular influx of Ca(2+). Both mouse aortic mesenchymal progenitors and mouse macrophage cells express POMC and the prohormone convertase 1/3 (PC1/3) indicating they have the potential to contribute to the local production of POMC peptides. These data demonstrate functional MC2-R expression in mouse aorta-derived mesenchymal progenitors and implicate both macrophage and mesenchymal cells as relevant sources of local POMC peptides.     

间充质干细胞治疗支气管肺发育不良的研究进展

支气管肺发育不良( bronchopulmonary dysplasia,BPD)常见于早产儿。20世纪80年代以来随着医疗技术的提高,早产儿成活率逐渐升高,同时BPD发病率也逐渐增加。目前,BPD的常规治疗并不理想,故亟需寻求一种新创性疗法减轻BPD造成的呼吸系统损伤,提高生活质量。近年来,间充质干细胞( mesenchymal stem cell,MSC)的研究为BPD的治疗提供了新切入点。该文就MSC治疗BPD的可能作用机制及目前研究现状进行了初步探讨。     

Paracrine factors from mesenchymal stem cells: a proposed therapeutic tool for acute lung injury and acute respiratory distress syndrome

Despite extensive researches in acute lung injury (ALI) and acute respiratory distress syndrome (ARDS), current pharmacological therapies and respiratory support are still the main methods to treat patients with ALI and ARDS and the effects remain limited. Hence, innovative therapies are needed to decrease the morbidity and mortality. Because of the proven therapeutic effects in other fields, mesenchymal stem cells (MSCs) might be considered as a promising alternative to treat ALI and ARDS. Numerous documents demonstrate that MSCs can exert multiple functions, such as engraftment, differentiation and immunoregulation, but now the key researches are concentrated on paracrine factors secreted by MSCs that can mediate endothelial and epithelial permeability, increase alveolar fluid clearance and other potential mechanisms. This review aimed to review the current researches in terms of the effects of MSCs on ALI and ARDS and to analyse these paracrine factors, as well as to predict the potential directions and challenges of the application in this field.     

LNG对双室培养的卵巢颗粒细胞和卵泡内膜细胞分泌功能的影响

本文采用羊膜双室培养系统,观察了18甲基炔诺酮(LNG)对猪卵巢颗粒细胞(G-C)和卵泡内膜细胞(T-C)在甾体激素生成过程中的影响。生长在羊膜两侧的G-C和T-C在加入或不加FSH、LH及各种不同浓度LNG的条件下孵育48小时,用RIA测定内、外室培养液中孕酮(P)和雌二醇(E_2)的含量,并与单独培养时的结果相比较。结果表明:①在FSH刺激时,LNG(30、3000nmol/L)明显抑制双室培养的G-C的P和E_2产量:P产量由55.1±3.4μmol/L降为25.8±1.8和20.3±3.8μmol/L;E_2产量由9.35±0.06nmol/L降至5.24±0.64和3.34±0.72nmol/L。但对P和E_2的基础水平无影响。②不论有或无LH刺激,LNG(30、3000nmoL/L)均明显抑制双室培养的T-C的P产量。有LH刺激时,P产量由70.9±6.5μmoL/L分别降为47.1±11.8和4.8±0.5μmol/L;在无LH刺激时,则由26.9±1.7μmol/L分别降至16.9±1.1和5.6±0.9μmol/L。结论:①LNG抑制双室培养中G-C的P和E_2产量,这种抑制作用是通过降低促性腺激素的刺激作用而产生的;②LNG不但抑制T-C的P基础分泌量,而且还表现为降低促性腺激素对P的刺激效应。③双室培养系统模拟了两种卵巢细胞在体时的旁分泌调节作用,与单独培养相比,是研究避孕药对卵巢功能影响的一种更为理想的模型。     

Divergent effect of glucagon antibodies on arginine and glucose-stimulated insulin secretion in the rat

Summary The effects of glucose and arginine on insulin secretion in the presence of glucagon antibodies were investigated in rats in vivo. In contrast to controls, animals given glucagon antibodies showed an inhibition of arginine-stimulated (p < 0.001), but not glucose-stimulated, insulin secretion. That these effects were not due to incomplete neutralisation of endogenous glucagon is evidenced by the presence of large antibody excess throughout the duration of the experiments. Both the glucagonotropic effect of arginine (319 ± 60ng/l, p < 0.01) and the insulinotropic effect of exogenous glucagon (8.3 ± 0.8 g/l, p < 0.001) were demonstrable under our experimental conditions in the absence of exogenous glucagon antibodies. These observations suggest that different mechanisms are involved in the stimulation of insulin release by arginine and by glucose, and that glucagon may play an important physiological role in the mediation and regulation of insulin secretion by secretogogues, such as arginine.