吸入糖皮质激素对大鼠肺纤维化的干预作用及bFGF表达的影响

目的:研究吸入糖皮质激素对大鼠肺纤维化模型的干预作用及可能的机制。方法:雌性Wistar大鼠40只,体重180～250g,按照随机数字表法将大鼠随机分为4组(n=10):①对照组(C组);②模型组(M组);③布地奈德组(B组);④地塞米松组(D组)。M组、B组、D组给大鼠气管内吸入博莱霉素(5mg/kgbw,8mg)复制肺纤维化模型,C组气管内吸入同等剂量的生理盐水作对照,B组于次日给予雾化吸入等效剂量布地奈德,D组于次日腹腔内注射地塞米松。上述各组均于注药后第1、4周各宰杀5只。通过苏木素-伊红染色观察肺泡炎、Masson胶原染色观察肺纤维化、用免疫组化及酶联免疫吸附测定(ELISA)法检测bFGF蛋白在大鼠肺组织,血清及肺泡灌洗液(BALF)的表达。结果:1、4周时M组表现为肺泡炎及肺间质炎症,B组、D组肺泡炎及肺纤维化程度较M组减轻。1、4周时血清、肺组织、BALF中M组的bFGF表达高于C组(P0.01),B、D组低于M组(P0.01)。结论:吸入糖皮质激素可减轻博莱霉素诱导的肺纤维化,其抗纤维化作用的机制与抑制bFGF表达有关。     

人源性抗bFGF单链抗体表达载体的构建与酵母表达

为了提高人源性抗bFGF抗体的表达量,从噬菌体抗体库筛选出的人源性抗bFGF抗体基因中亚克隆单链抗体(single chain fragment variable,ScFv)基因,并将其构建到酵母表达载体pPICZαA中。表达载体质粒经线性化后,电转化法转化至毕赤酵母GS115中,甲醇诱导表达。表达产物经镍离子亲和层析和阴离子交换层析纯化,并检测其生物学活性。酶切鉴定结果显示人源性的酵母表达载体构建成功。SDS-PAGE和Western blot结果显示,抗bFGF单链抗体获得了高效表达,表达量可达124mg/L,目的蛋白大小为36 kDa左右。通过两步纯化方案,目的蛋白的纯度可达95%以上。ELISA结果显示纯化的目的蛋白可与bFGF特异性结合。CCK8检测结果显示,纯化的抗bFGF单链抗体可剂量依赖性地抑制人肺腺癌细胞株A549的增殖。研究结果表明在毕赤酵母中可获得人源性抗bFGF单链抗体高效表达,表达产物具有很好的生物学活性。     

脱落酸抗体的抗独特型抗体的制备与鉴定

用ABA多克隆抗体(Ab_1)以金黄色葡萄球菌菌体(SPA)作为载体,免疫家兔,制备的抗独特型抗体(Ab_2)初步纯化后用ELISA试验鉴定其特异性的结果表明,该抗独特型抗体具有良好的特异性,能阻断ABA抗体对ABA的结合反应,并与ABA结合蛋白结合,暗示其有模拟抗原的作用。 Jerne的免疫网络学说认为,特异性抗体(Ab_1)可变区中的独特型决定簇(Id)可诱导抗独特型抗体(Ab_2)的产生,Ab_2中的一部分可以模拟抗原的作用。Sege和Peterson提出,     

三七总皂甙对博莱霉素所致小鼠肺纤维化的干预作用

目的:观察三七总皂甙(PNS)对实验性小鼠肺纤维化的干预作用。方法:小鼠随机分为正常对照组、模型对照组、醋酸泼尼松组和PNS大、中、小剂量组。通过气管内注入博莱霉素(BLM)复制小鼠肺纤维化模型,于造模后第2天各治疗组开始给药,于给药后第7、14、28 d处死部分小鼠,取肺组织,行HE染色,并测定肺组织中羟脯氨酸(HYP)含量。结果PNS能减少实验性肺纤维化小鼠肺组织中胶原沉积及降低肺系数(P<0.05,P<0.01),减轻肺部的病理损害(P<0.05,P<0.01)。结论:PNS对BLM诱导产生的小鼠肺纤维化有一定的抑制作用。     

抗中性粒细胞胞浆抗体、抗核抗体联合检测对类风湿关节炎的诊断价值

目的：探讨抗中性粒细胞胞浆抗体（ANCA）与抗核抗体（ANA）联合检测对类风湿关节炎的临床意义。方法：采用IIF法对82例RA患者（RA组）、74例非RA自身免疫疾病患者（非RA组）和52例健康体检者（正常对照组）的血清ANCA和ANA谱进行了检测分析,并用ELISA法进行抗丝氨酸蛋白酶3（PR3）、抗髓过氧化物酶（MPO）、ANA谱的定量检测。结果：RA组82例患者中,64例ANCA阳性,阳性率为78．08％,其中核周型（PANCA）37例,阳性率为45．1％,胞浆型（CANCA）27例,阳性率为32．9％;非RA组74例患者中有7例ANCA阳性率分别为9．4％;正常对照组50例中没有一例ANCA阳性。利用Elisa法对患者血清进行检测,分别能够特异的检测到PR3、MPO、抗双链DNA抗体（抗ds—DNA抗体）、抗SS—A等抗体、抗ss—A抗体、抗PM—SCL抗体的存在。结论：联合ANCA、ANA检测有助于提高类风湿关节炎的诊断。     

抗中性粒细胞胞浆抗体、抗核抗体联合检测对类风湿关节炎 的诊断价值

目的:探讨抗中性粒细胞胞浆抗体(ANCA)与抗核抗体(ANA)联合检测对类风湿关节炎的临床意义。方法:采用IIF法对82例RA患者(RA组)、74例非RA自身免疫疾病患者(非RA组)和52例健康体检者(正常对照组)的血清ANCA和ANA谱进行了检测分析,并用ELISA法进行抗丝氨酸蛋白酶3(PR3)、抗髓过氧化物酶(MPO)、ANA谱的定量检测。结果:RA组82例患者中,64例ANCA阳性,阳性率为78.08%,其中核周型(PANCA)37例,阳性率为45.1%,胞浆型(CANCA)27例,阳性率为32.9%;非RA组74例患者中有7例ANCA阳性率分别为9.4%;正常对照组50例中没有一例ANCA阳性。利用Elisa法对患者血清进行检测,分别能够特异的检测到PR3、MPO、抗双链DNA抗体(抗ds-DNA抗体)、抗ss-A等抗体、抗SS-A抗体、抗PM-SCL抗体的存在。结论:联合ANCA、ANA检测有助于提高类风湿关节炎的诊断。     

慢性抗Thy1抗体肾炎模型的研究进展

慢性抗thy1抗体肾炎模型由于其疾病过程与人类IgA肾病及其他的系膜增生性肾炎的疾病进程相似,可以用作研究慢性肾脏病的发病机制以及新的治疗策略,本文就慢性抗thy1抗体肾炎模型的建立方法、病理演变过程、发病机制及治疗进展做一综述.     

虎杖苷抗百草枯中毒肺纤维化作用及机制的研究进展

百草枯(Paraquat,PQ)对人畜均有很强的毒性,可引起肺组织不可逆的弥漫性纤维化,最终导致患者因呼吸衰竭而死亡。虎杖苷(Polydatin,PD)是蓼科植物虎杖的干燥根茎中分离出来的一种芪类有机化合物,也是其发挥药理作用的主要成分,具有抗氧化、抗肿瘤、抗炎、清除自由基、降低肺纤维化进程等丰富的药理活性。目前研究表明PD治疗PQ中毒肺纤维化比较明确的机理可归纳为抗氧化应激、调节细胞因子网络调控失衡以及抗基质金属蛋白酶(MMPs)/金属蛋白酶组织抑制因子(TIMPs)的失衡。本文主要对PD抗PQ中毒肺纤维化作用机制的研究进展进行了综述。     

罗格列酮对肺纤维化大鼠肺动脉壁结缔组织生长因子表达的影响

目的:观察过氧化物酶体增殖活化受体γ(PPAR-γ)激动剂罗格列酮(RSG)对肺纤维化大鼠肺动脉壁结缔组织生长因子(CTGF)上调、Ⅰ型和Ⅲ型胶原沉积的影响。方法:48只雄性SD大鼠,随机分为以下4组:博莱霉素(BLM)+生理盐水(NS)组(n=21)、BLM+RSG组(n=9)、NS+NS组(n=9)和NS+RSG组(n=9)。气管内一次性滴注BLM(5mg/kgbw),RSG灌胃(3mg/(kg.d),14d)。整体实验,气管滴注后第14天观察;离体实验,气管滴注BLM后第14天,分离大鼠的肺动脉,并用RSG培养液和单纯培养液孵育(37℃,5%CO2,24h)。结果:在整体水平,与对照大鼠相比,BLM模型大鼠肺动脉壁的CTGF免疫阳性表达增强,CTGF蛋白含量、Ⅰ型和Ⅲ型胶原含量、Ⅰ/Ⅲ胶原比值均增高(均P0.05);RSG能阻止上述指标的异常变化(均P0.05);在离体水平,RSG能阻止BLM模型大鼠肺动脉壁CTGF的上调(P0.05),但对Ⅰ型和Ⅲ型胶原沉积无明显影响(P0.05)。结论:RSG能直接作用于肺动脉壁,阻止肺纤维化大鼠肺动脉壁CTGF的上调,这可能是其减轻动脉壁结构重塑的机制之一。     

Effects of fish oil treatment on bleomycin-induced pulmonary fibrosis in mice

Bleomycin is an antibiotic used to treat a variety of neoplasms. A major side-effect of bleomycin therapy is the induction of an intense inflammatory response that develops into pulmonary fibrosis. Several studies have shown that certain polyunsaturated fatty acids found in fish oil reduce the inflammatory response in vivo. Fish oil has been employed for the treatment of several pathologies such as glomerulonephritis, cardiovascular diseases, rheumatoid arthritis, and even as an adjuvant in cancer therapy. This study examined the effects of fish oil treatment on the development of bleomycin-induced pulmonary fibrosis. Mice were intraperitoneally treated with bleomycin or with saline daily for 10 days, and 15 days after the last injection they started to receive fish oil by gavage for 14 days. The lungs were processed for light microscopy, biochemical and immunohistochemical investigations. Fish oil did not prevent the development of pulmonary fibrosis after the injury as shown by light microscopy, cytokines immunohistochemical analysis, TBARS content and protein levels in the lung. In addition however, fish oil itself induced a slight inflammatory process in the lung, as observed by the increase in cellularity, vasodilatation in the lung parenchyma, TBARS content, and a slight increase in the lung protein content.     

Orai3 mediates Orai channel remodelling to activate fibroblast in pulmonary fibrosis

Orai family are a calcium channel of cell membrane extracellular Ca²⁺ influx which participates in tissue fibrosis. But the roles of Orai3 have less attention on the mechanism of regulating lung fibrosis. In this study, we found that Orai3 expression was increased significantly in BLM‐induced lung fibrosis. The knockdown of Orai3 decreased TGF‐β1‐induced fibroblast proliferation, ECM production, activation of NFAT1 and Calpain/ERK signal pathway and glycolysis levels. Orai3 interacting with Orai1 was increased in BLM‐induced lung fibrosis and TGF‐β1‐induced fibroblast, while the Stim1 interacting with Orai1 and SOCE activity was suppressed, leading in a high and stable extracellular Ca²⁺ influx. Furthermore, the over‐expression of Orai3 did not enhance Orai3 interacting with Orai1 under TGF‐β1 free fibroblast. And then, the deeper mechanism of TGF‐β1‐induced increased SEPTIN4 promoted Orai3 interacting with Orai1. Our results indicated that Orai3 could be one of the therapy targets for PF in which remodels Orai channel, suppresses SOCE activity and activated fibroblast to alleviate fibrosis progress.     

Silencing fibroblast growth factor 7 inhibits krypton laser-induced choroidal neovascularization in a rat model

Choroidal neovascularization (CNV), a characteristic of age-related macular degeneration, is an underlying cause of severe vision loss among elderly patients. Fibroblast growth factor (FGF) is suggested to exert an important role in the pathogenesis of CNV. However, the molecular mechanisms governing this event are not fully elucidated. Herein, we identified the potential role of FGF7 in CNV. To examine the roles of FGF7 in the progression of CNV, rat CNV models were established and treated with small interfering RNA (siRNA) against FGF7 or FGF7 overexpression, followed by identification of expression of FGF7 in the CNV modeled rats. Next, proliferation and migration, and in vitro tube formation of human umbilical vein endothelial cells, as well as expression of vascular endothelial growth factor (VEGF) and transforming growth factor-beta 2 (TGF-β2) were evaluated. CNV led to upregulated FGF7 expression. Cells in the presence of FGF7 siRNA showed suppressed proliferation, migration, and tube formation, along with downregulated VEGF and TGF-β2 expression. Taken together, functional suppression of FGF7 inhibited the onset of CNV, ultimately highlighting a novel therapeutic target for suppressing CNV progression.     

Polydatin alleviates bleomycin-induced pulmonary fibrosis and alters the gut microbiota in a mouse model

To investigate the effect and mechanism of polydatin on bleomycin (BLM)-induced pulmonary fibrosis in a mouse model. The lung fibrosis model was induced by BLM. The contents of TNF-α, LPS, IL-6 and IL-1β in lung tissue, intestine and serum were detected by ELISA. Gut microbiota diversity was detected by 16S rDNA sequencing; R language was used to analyse species composition, α-diversity, β-diversity, species differences and marker species. Mice were fed drinking water mixed with four antibiotics (ampicillin, neomycin, metronidazole, vancomycin; antibiotics, ABx) to build a mouse model of ABx-induced bacterial depletion; and faecal microbiota from different groups were transplanted into BLM-treated or untreated ABx mice. The histopathological changes and collagen I and α-SMA expression were determined. Polydatin effectively reduced the degree of fibrosis in a BLM-induced pulmonary fibrosis mouse model; BLM and/or polydatin affected the abundance of the dominant gut microbiota in mice. Moreover, faecal microbiota transplantation (FMT) from polydatin-treated BLM mice effectively alleviated lung fibrosis in BLM-treated ABx mice compared with FMT from BLM mice. Polydatin can reduce fibrosis and inflammation in a BLM-induced mouse pulmonary fibrosis model. The alteration of gut microbiota by polydatin may be involved in the therapeutic effect.     

大鼠肺纤维化形成中肺内肥大细胞结缔组织生长因子的表达

目的：观察博莱霉素（BLM）诱导肺纤维化形成中肺肥大细胞（MCs）是否表达结缔组织生长因子（CTGF）。方法：32只雄性SD大鼠,随机分为博莱霉素（BLM）组和对照（Control）组（n=16）。BLM组为气管内一次性滴注BLM（5mg／ks）;Control组为气管内滴注与BLM等容量的生理盐水（NS）。各组分别在气管滴注后第14天和第28天处死大鼠,取肺组织样本。用氯胺-T法检测肺组织羟脯氨酸含量以判断肺纤维化程度;用甲苯胺蓝染色显示肺组织切片中的MCs;免疫组化染色显示肺CTGF的表达和分布。结果：①与对照大鼠比,气管内滴注BLM后第28天大鼠的肺羟脯氨酸含量明显增高（P〈0．01）。②与对照大鼠比,气管内滴注BLM后第14天和第28天大鼠肺内MCs数明显增多（均P〈0．01）,肺内CTGF表达上调（均P〈0．01）。③对照大鼠肺内未见CTGF免疫阳性的MCs;而气管内滴注BLM后第14天和第28天大鼠肺内病灶区中有CTGF免疫阳性的MCs。结论：肺纤维化形成中肺MCs表达CTGF,这可能是MCs促进肺纤维化的作用机制之一。