百草枯对A549细胞中解整合素-金属蛋白酶17表达的影响

目的构建体外百草枯（paraquat,PQ）细胞纤维化模型,观察PQ对A549细胞中解整合素-金属蛋白酶17（ADAM17）表达的影响,探讨ADAM17在PQ中毒致肺纤维化中的作用。方法体外培养A549细胞,分为正常对照组、不同浓度PQ组,应用CCK-8检测细胞活力,筛选PQ浓度和时间,显微镜下观察细胞形态,ELISA测定各组纤维化标志物I型胶原（type I collagen,Col I）和纤连蛋白（fibronectin,FN）的表达,建立细胞纤维化模型;免疫细胞化学检测A549细胞中ADAM17的分布情况,RT-PCR和Western blot分别半定量检测ADAM17 mRNA及蛋白水平的表达情况。结果（1）随着PQ浓度增加及作用时间延长,A549细胞活力呈下降趋势,差异有统计学意义（P〈0.05）,呈剂量依赖性和时间依赖性。（2）正常的A549细胞融合呈铺路石样生长,排列比较紧密,经PQ诱导后细胞排列较松散,细胞间连接变疏松,部分细胞溶解、死亡。（3）ELISA显示,随PQ浓度增加,Col I和FN表达增强,差异有统计学意义（P〈0.05）;随PQ时间延长,Col I和FN表达也逐渐增强,差异有统计学意义（P〈0.05）,成功建立PQ细胞纤维化模型。（4）免疫细胞化学显示,ADAM17在A549细胞胞浆表达。（5）RT-PCR和Western blot表明,随着PQ浓度增加,ADAM17 mRNA及蛋白水平表达明显增加,差异有统计学意义（P〈0.05）,以PQ 200 umol/L时最为明显。随着PQ作用时间延长,ADAM17 mRNA及蛋白表达水平也明显增加,差异有统计学意义（P〈0.05）,在24 h达到高峰。结论百草枯可引起肺泡上皮细胞形态学改变,导致细胞损伤,成功建立细胞的纤维化模型,对A549细胞的毒性作用具有剂量和时间依赖性。ADAM17在PQ诱导的A549细胞中过表达,可能参与了百草枯诱导的肺纤维化过程。     

Protective effect of MOTILIPERM in varicocele-induced oxidative injury in rat testis by activating phosphorylated inositol requiring kinase 1α (p-IRE1α) and phosphorylated c-Jun N-terminal kinase (p-JNK) pathways

Context: MOTILIPERM was prepared as a mixture of extracts of three medicinal herbs [roots of Morinda officinalis How (Rubiaceae), outer scales of Allium cepa L. (Liliaceae) and seeds of Cuscuta chinensis Lamark (Convolvulaceae)].

Objective: To investigate the role of reactive oxygen species (ROS)-based endoplasmic reticulum (ER) stress in a rat model of varicocele and the therapeutic efficacy of MOTILIPERM in this model.

Materials and methods: Sixty male rats were divided into five experimental groups: a normal control group (CTR?+?vehicle), a control group administered MOTILIPERM 200?mg/kg (CTR?+?M 200), a varicocele-induced control group (VC?+?vehicle) and two varicocele-induced groups administered MOTILIPERM 100 (VC?+?M 100) or 200 (VC?+?M 200) mg/kg for 4 weeks. Testis weights were recorded and serums were assayed for hormone concentrations. Tissues were subjected to semen analysis, histopathology, analyses of ER response protein expression levels and oxidative stress were assessed by measuring ROS, reactive nitrogen species (RNS), malondialdehyde (MDA) level and ratios of total glutathione (GSH)/oxidized GSH (GSSG).

Results: MOTILIPERM treatment of varicocele-induced groups significantly increased left testis weight, testosterone level, sperm motility, count and spermatogenic cell density. ER-response protein expression levels were dose-dependently decreased in VC?+?M 200 group compared with VC?+?vehicle group. MOTILIPERM treatment also decreased MDA and ROS/RNS level but increased GSH/GSSG ratio.

Discussion and conclusions: This study suggests that ROS-related ER stress may play a major role in varicocele-induced infertility and MOTILIPERM, a novel compound targeting ROS-based ER stress, may be therapeutically useful in treatment of varicocele, or as a supplement for the treatment of infertility.     

A visible fluorescent nanovaccine based on functional genipin crosslinked ovalbumin protein nanoparticles

Accurate and efficient antigen delivery is crucial for inducing a strong and long-term immune response. A visible protein nanovaccine made from antigen could provide a novel and promising technology for secure and efficient delivery of the antigen with imaging visualization. In this study, a functional nanovaccine based on genipin crosslinked ovalbumin (OVA) fluorescent nanoparticles with chitosan (CS-OVA-NPs) was developed. The nanovaccine can carry abundant antigens by self-crosslinking without additional carriers. The fluorescence imaging technique was applied to monitor and reveal the process of antigen delivery in vivo based on the fluorescence of genipin with a non-invasive and real-time manner. This functional OVA nanovaccine can enhance the uptake of OVA in Dendritic Cells (DCs) and further promote DCs to maturate in vitro. In vivo study further indicated CS-OVA-NPs could trigger antigen-specific immune responses, which demonstrated that this fluorescent nanovaccine provided a novel design approach for accurate and efficient vaccine delivery.