Effect of sciatectomy and induced ammonia stress on Mg2+_adenosine triphosphatase activity in frog tissues

Mg² + dependent —adenosine triphosphatase activity has been studied in the muscle, brain, kidney and liver tissues of frog,Rana hexadactyla (Lesson) after sciatectomy and induced chronic ammonia stress. The enzyme activity decreased in the tissues of the denervated frog. The activity of the enzyme increased in all the tissues of the normal and denervated frogs except in the denervated muscle when ammonium lactate was infused intraperitoneally.     

An arachidonic acid generation/export system involved in the regulation of cholesterol transport in mitochondria of steroidogenic cells

Recent studies demonstrated the importance of the mitochondrial ATP in the regulation of a novel long-chain fatty acid generation/export system in mitochondria of diabetic rat heart. In steroidogenic systems, mitochondrial ATP and intramitochondrial arachidonic acid (AA) generation are important for steroidogenesis. Here, we report that mitochondrial ATP is necessary for the generation and export of AA, steroid production and steroidogenic acute regulatory protein induction supported by cyclic 3'-5'-adenosine monophosphate in steroidogenic cells. These results demonstrate that ATP depletion affects AA export and provide new evidence of the existence of the fatty acid generation and export system involved in mitochondrial cholesterol transport.     

Particulate matter 2.5 induced bronchial epithelial cell injury via activation of 5′-adenosine monophosphate-activated protein kinase-mediated autophagy

The impact of particulate matter 2.5 (PM2.5) on the respiratory system is a worldwide concern. However, the mechanisms by which PM2.5 causes disease are still unclear. In this study, we investigated the effect of PM2.5 on autophagy and studied the effect of PM2.5-induced autophagy and 5′-adenosine monophosphate-activated protein kinase (AMPK) on cell proliferation, cell cycle, apoptosis, reactive oxygen species (ROS), and airway inflammation using human bronchial epithelial cells 16HBE140 cells. Results showed that exposure of cells to PM2.5 at a concentration of 100 μg/mL for 24 hours was most effective for inhibiting cell viability. PM2.5 induced cell arrest in the G0/G1 phase and increased mitochondrial membrane potential, ROS, and cell apoptosis with increasing concentration. PM2.5 downregulated cyclin D and matrix metallopeptidase-9 (MMP-9) expression but upregulated tissue inhibitor of metalloproteinases-1 (TIMP-1) expression, significantly promoted interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) production, and enhanced the level and activation of AMPK. The levels of autophagy-related protein 5 (ATG5), Beclin-1, and LC3II/I were significantly increased by PM2.5. The activation of Unc-51-like autophagy activating kinase 1 was significantly inhibited by PM2.5. Moreover, ATG5 knockdown inhibited PM2.5-induced autophagy, ROS, and cell apoptosis significantly. The expression of cyclin D, MMP-9, and TIMP-1 was reversed by ATG5 suppression. PM2.5-induction of IL-6 and TNF-α was significantly inhibited by knockdown of ATG5. Thus, inhibition of autophagy protected the cells from PM2.5-induced injury. PM2.5 induced injury in human bronchial epithelial cells via activation of AMPK-mediated autophagy, suggesting possible therapeutic targets for the treatment of respiratory diseases.     

Metformin protects against doxorubicin-induced cardiotoxicity: involvement of the adiponectin cardiac system

Doxorubicin has cardiotoxic effects that limit its clinical benefit in cancer patients. Metformin exerts cardioprotective actions via AMP-activated protein kinase (AMPK) and increases the expression of adiponectin and its receptors (adipoR1 and adipoR2) in skeletal muscle and adipose tissue, but its effect on cardiac tissue is still unknown. This work aimed to study whether metformin exerts any protective action against the cardiotoxicity of doxorubicin and whether the cardiac system of adiponectin is involved in any such action. The addition of doxorubicin (5μM) to adult mouse cardiomyocytes (HL-1 cell line) induced apoptosis, which was characterized by a loss of cell viability, activation of caspases, and fragmentation of the genetic material. Doxorubicin treatment also caused a decrease in the activity of the antioxidant enzymes catalase, glutathione peroxidase, and superoxide dismutase. Pretreatment with metformin (4mM, 24h) provided protection against doxorubicin-induced damage. This pretreatment significantly increased cell viability, attenuated the activation of caspases and the fragmentation of genetic material, and restored the antioxidant activity. In addition, metformin up-regulated the expression of adiponectin and its receptors, adipoR1 and adipoR2, in cardiomyocytes. In contrast, silencing either adipoR1 or adipoR2 with siRNA inhibited the AMPK activation and the protective effects of metformin. Taken together, these results demonstrate that metformin protects cardiomyocytes from doxorubicin-induced damage and that the cardiac adiponectin system plays an important role in this protective action.     

5′-O-Fluorosulfonylbenzoyl Esters of Purine Nucleosides as Potential Inhibitors of NTPase/Helicase and Polymerase of Flaviviridae Viruses

Abstract

Synthesis and interactions of guanosine, inosine and ribavirin 5′-fluorosulfonyl-benzoyl esters with hepatitis C virus (HCV) and Flaviviruses NTPase/helicase and polymerase are described.     

粉被虫草静置发酵产N6-(2-羟乙基)腺苷培养基组分及前体物的筛选

在静置培养条件下,对粉被虫草cp菌株产N6-(2-羟乙基)腺苷（简称HEA）的培养基及其组分进行优化。采用单因素实验方法,以HPLC法对cp菌丝体中HEA进行检测,以HEA的产量为指标,结果表明查氏培养基有利于cp菌株产HEA,其产量是沙氏和PD培养基的3.7倍和4.48倍。以查氏培养基为基础培养基,进行碳、氮源的筛选中,最有利于cp菌株产HEA的是蔗糖和硝酸钠;在无机盐的筛选中,K2HPO4是促进cp菌株累积HEA最主要的无机盐,其HEA产量较CK增长了5 822.37%（提高了58.2倍）;在查氏培养基上添加不同前体物和氨基酸,发现其结构类似物腺苷、次黄嘌呤和腺嘌呤都有促进cp菌株累积HEA的能力,其中腺苷和次黄嘌呤能提高其产量60%以上;而组氨酸是最有利于cp菌株累积HEA的氨基酸,产量达到（45.56±2.8）mg/L,较CK提高HEA产量251.68%,其次是L-谷氨酸增产184.19%。     

蝉花虫草活性成分的抗惊厥作用

本研究采用活性追踪的方法,逐步从人工培养蝉花虫草分离获得A（50%乙醇回流提取）、B（膜分离）、C（大孔树脂洗脱）和D（Sephadex LH20柱纯化）等4种样品,单一化合物D纯度为98.62%,鉴定为N6‐(2‐羟乙基)腺苷[N6‐(2‐hydroxyethyl)‐adenosine,HEA]。研究各样品对戊四唑（pentylenetetrazol,PTZ）诱导的小鼠惊厥模型的影响,以及选择性腺苷A1受体（AA1R）拮抗剂DPCPX或选择性腺苷A2A受体（AA2AR）拮抗剂ZM241385对HEA作用的影响,并采用苏木精-伊红（hematoxylin-eosin,HE）染色、免疫组化（immunohistochemical,IHC）染色和蛋白质免疫印迹（Western blot,WB）等技术进一步探究HEA抗惊厥作用的机制。结果表明,各样品（i.p.）均有抗惊厥活性,HEA（40–60mg/kg,i.p.）能显著延长惊厥小鼠的存活时间和降低死亡率,DPCPX（2mg/kg,i.p.）能够拮抗HEA的抗惊厥作用,而ZM241385无此作用;HE、IHC和WB的结果进一步揭示DPCPX显著降低HEA的作用。综上所述,蝉花虫草的HEA具有抗惊厥作用,并且可能通过激活腺苷A1受体而起作用。