Astaxanthin Alleviates Early Brain Injury Following Subarachnoid Hemorrhage in Rats: Possible Involvement of Akt/Bad Signaling

Apoptosis has been proven to play a crucial role in early brain injury pathogenesis and to represent a target for the treatment of subarachnoid hemorrhage (SAH). Previously, we demonstrated that astaxanthin (ATX) administration markedly reduced neuronal apoptosis in the early period after SAH. However, the underlying molecular mechanisms remain obscure. In the present study, we tried to investigate whether ATX administration is associated with the phosphatidylinositol 3-kinase-Akt (PI3K/Akt) pathway, which can play an important role in the signaling of apoptosis. Our results showed that post-SAH treatment with ATX could cause a significant increase of phosphorylated Akt and Bad levels, along with a significant decrease of cleaved caspase-3 levels in the cortex after SAH. In addition to the reduced neuronal apoptosis, treatment with ATX could also significantly reduce secondary brain injury characterized by neurological dysfunction, cerebral edema and blood-brain barrier disruption. In contrast, the PI3K/Akt inhibitor, {"type":"entrez-nucleotide","attrs":{"text":"LY294002","term_id":"1257998346","term_text":"LY294002"}}LY294002, could partially reverse the neuroprotection of ATX in the early period after SAH by downregulating ATX-induced activation of Akt/Bad and upregulating cleaved caspase-3 levels. These results provided the evidence that ATX could attenuate apoptosis in a rat SAH model, potentially, in part, through modulating the Akt/Bad pathway.     

Caspase-9在肉用绵羊主要生殖器官中的分布研究

通过试验研究凋亡因子Caspase-9在肉用绵羊主要生殖器官中的表达,并探讨其生理意义.利用免疫组织化学的方法对Caspase-9在肉用绵羊主要生殖器官中的表达进行研究.研究结果显示,Caspase-9只在细胞质中表达,其在卵巢中主要分布于原始卵泡、初级卵泡及颗粒性黄体细胞,而卵泡膜性黄体细胞中未见表达;黄体期,主要表达于子宫的浅层腺体腺上皮细胞、子宫内膜上皮细胞、子宫颈黏膜固有层的淋巴小结、输卵管黏膜上皮分泌细胞和纤毛细胞、峡部的浆液性腺的腺上皮细胞;卵泡期,子宫和输卵管各个部位阳性反应不明显.在正常生理情况下,Caspase-9参与了肉用绵羊主要生殖器官周期性变化的调控和子宫黏膜免疫,对生殖器官功能的稳定发挥起着重要作用.     

CPEB3 regulates the proliferation and apoptosis of bovine cumulus cells

Cytoplasmic polyadenylation element‐binding protein 3 (CPEB3) is a member of the Cytoplasmic polyadenylation element‐binding family, which has been found to regulate the translation of dormant and masked mRNA in Xenopus oocytes and plays potential roles in regulating biological functions in cells and tissues. However, its role in cumulus cells is not clear. In this study, the mRNA expression of CPEB3 in bovine cumulus cells was inhibited with small interfering RNA. Cell cycle progression, proliferation, and apoptosis were measured after inhibition of CPEB3. Subsequently, changes in intracellular Reactive oxygen species content, mitochondrial membrane potential and expansion‐related gene expression were examined. The results showed that after CPEB3 inhibition, cumulus cells had an abnormal cell cycle, the numbers of cells in the S and G2/M phases were significantly increased, cell proliferation was increased and apoptosis rates were decreased. These effects were likely due CPEB3 inhibition‐induced decreases in intracellular Reactive oxygen species levels; increases in mitochondrial membrane potential; decreases in apoptosis; downregulation of CCNA, CCND, CCNE, CDK2, CDK4, CDK6, p21, and p27 mRNA expression; and upregulation of CCNB, CDK1, HAS2, PTGS2, PTX3, and CEBPB mRNA expression. Therefore, CPEB3 plays potential roles in regulating the biological and physiological functions of bovine cumulus cell.     

热应激对鸡胸腺细胞凋亡的影响及其调节机理

18只AA肉鸡于 2 2℃饲养。 30日龄时 ,随机分成 3组 ,分别进行 0 ,5和 10h的 4 1℃高温应激处理。用流式细胞仪检测胸腺细胞DNA含量、细胞凋亡率以及细胞caspase 3和Bcl 2的表达。结果 :应激处理 0 ,5和 10h时 ,(1)DNA合成期 (S期 )细胞比例分别是 2 77% ,3 2 8%和 2 31% ;(2 )胸腺细胞凋亡率分别是 3 99% ,11 15 %和3 31% ;(3)caspase 3表达阳性细胞率分别为 14 2 4 % ,2 7 0 1%和 10 84 % ,Bcl 2表达阳性细胞率分别是 2 0 5 9% ,15 13%和 2 7 92 % ;(4 )应激 5h与 0和 10h的细胞凋亡率、caspase 3和Bcl 2的阳性细胞表达率均有显著差异 (P <0 0 5 ) ;(5 )应激处理鸡的胸腺细胞凋亡率、caspase 3表达阳性细胞率从 0h到 5h逐渐升高 ,以后又逐渐下降。在高温应激过程中 ,胸腺细胞凋亡率与caspase 3表达阳性细胞率呈平行的变化关系 ,而与Bcl 2表达阳性细胞率呈反向变化关系。这些结果均说明 :高温应激能明显影响胸腺细胞凋亡 ,而这种细胞凋亡的变化受caspase 3和Bcl 2表达的调节。     

The Inhibition Effect of the Seaweed Polyphenol, 7-Phloro-Eckol from Ecklonia Cava on Alcohol-Induced Oxidative Stress in HepG2/CYP2E1 Cells

The liver is vulnerable to oxidative stress-induced damage, which leads to many diseases, including alcoholic liver disease (ALD). Liver disease endanger people’s health, and the incidence of ALD is increasing; therefore, prevention is very important. 7-phloro-eckol (7PE) is a seaweed polyphenol, which was isolated from Ecklonia cava in a previous study. In this study, the antioxidative stress effect of 7PE on HepG2/CYP2E1 cells was evaluated by alcohol-induced cytotoxicity, DNA damage, and expression of related inflammation and apoptosis proteins. The results showed that 7PE caused alcohol-induced cytotoxicity to abate, reduced the amount of reactive oxygen species (ROS) and nitric oxide (NO), and effectively inhibited DNA damage in HepG2/CYP2E1 cells. Additionally, the expression levels of glutathione (GSH), superoxide dismutase (SOD), B cell lymphoma 2 (Bcl-2), and Akt increased, while γ-glutamyltransferase (GGT), Bcl-2 related x (Bax), cleaved caspase-3, cleaved caspase-9, nuclear factor-κB (NF-κB), and JNK decreased. Finally, molecular docking proved that 7PE could bind to BCL-2 and GSH protein. These results indicate that 7PE can alleviate the alcohol-induced oxidative stress injury of HepG2 cells and that 7PE may have a potential application prospect in the future development of antioxidants.     

Effect of Oversulfation on the Composition,Structure, and In Vitro Anti-Lung Cancer Activity of Fucoidans Extracted from Sargassum aquifolium

Intensive efforts have been undertaken in the fields of prevention, diagnosis, and therapy of lung cancer. Fucoidans exhibit a wide range of biological activities, which are dependent on the degree of sulfation, sulfation pattern, glycosidic branches, and molecular weight of fucoidan. The determination of oversulfation of fucoidan and its effect on anti-lung cancer activity and related signaling cascades is challenging. In this investigation, we used a previously developed fucoidan (SCA), which served as a native fucoidan, to generate two oversulfated fucoidan derivatives (SCA-S1 and SCA-S2). SCA, SCA-S1, and SCA-S2 showed differences in compositions and had the characteristic structural features of fucoidan by Fourier transform infrared (FTIR) and nuclear magnetic resonance (NMR) analyses. The anticancer properties of SCA, SCA-S1, and SCA-S2 against human lung carcinoma A-549 cells were analyzed in terms of cytotoxicity, cell cycle, Bcl-2 expression, mitochondrial membrane potential (MMP), expression of caspase-3, cytochrome c release, Annexin V/propidium iodide (PI) staining, DNA fragmentation, and the underlying signaling cascades. Our findings indicate that the oversulfation of fucoidan promotes apoptosis of lung cancer cells and the mechanism may involve the Akt/mTOR/S6 pathway. Further in vivo research is needed to establish the precise mechanism whereby oversulfated fucoidan mitigates the progression of lung cancer.     

Cracking the Cytotoxicity Code: Apoptotic Induction of 10-Acetylirciformonin B is Mediated through ROS Generation and Mitochondrial Dysfunction

A marine furanoterpenoid derivative, 10-acetylirciformonin B (10AB), was found to inhibit the proliferation of leukemia, hepatoma, and colon cancer cell lines, with selective and significant potency against leukemia cells. It induced DNA damage and apoptosis in leukemia HL 60 cells. To fully understand the mechanism behind the 10AB apoptotic induction against HL 60 cells, we extended our previous findings and further explored the precise molecular targets of 10AB. We found that the use of 10AB increased apoptosis by 8.9%–87.6% and caused disruption of mitochondrial membrane potential (MMP) by 15.2%–95.2% in a dose-dependent manner, as demonstrated by annexin-V/PI and JC-1 staining assays, respectively. Moreover, our findings indicated that the pretreatment of HL 60 cells with N-acetyl-l-cysteine (NAC), a reactive oxygen species (ROS) scavenger, diminished MMP disruption and apoptosis induced by 10AB, suggesting that ROS overproduction plays a crucial rule in the cytotoxic activity of 10AB. The results of a cell-free system assay indicated that 10AB could act as a topoisomerase catalytic inhibitor through the inhibition of topoisomerase IIα. On the protein level, the expression of the anti-apoptotic proteins Bcl-xL and Bcl-2, caspase inhibitors XIAP and survivin, as well as hexokinase II were inhibited by the use of 10AB. On the other hand, the expression of the pro-apoptotic protein Bax was increased after 10AB treatment. Taken together, our results suggest that 10AB-induced apoptosis is mediated through the overproduction of ROS and the disruption of mitochondrial metabolism.     

钼镉联合胁迫对山羊肾脏细胞凋亡相关基因表达的影响

本研究以波尔山羊为实验动物,旨在观察钼镉联合胁迫对山羊肾脏细胞凋亡相关基因表达的影响。选用63头健康波尔山羊随机分成7组,每组3个重复,选用七钼酸铵([(NH4)6Mo7O24·4H2O])作为实验钼源,氯化镉(Cd Cl2)作为实验镉源,按每kg山羊体质量添加钼、镉对照组(Mo 0 mg/kg+Cd 0 mg/kg),低镉低钼组(Mo 15mg/kg+Cd 0.5 mg/kg),低镉中钼组(Mo 30 mg/kg+Cd 0.5 mg/kg),低镉高钼组(Mo 45 mg/kg+Cd 0.5 mg/kg),高镉低钼组(Mo 15 mg/kg+Cd 1.0 mg/kg),高镉中钼组(Mo 30 mg/kg+Cd 1.0 mg/kg),高镉高钼组(Mo 45 mg/kg+Cd1.0 mg/kg),实验期50 d。并于当天、第25、50天每组各剖杀3头山羊取肾组织进行相关实验,观察不同剂量钼、镉对山羊肾脏细胞凋亡相关基因表达等的影响。结果显示,与对照组相比,在实验第25、50天时,实验组山羊肾脏bcl-2 mRNA表达量下降(P0.01),bax mRNA、caspase-3 mRNA和cytc mRNA表达量上升(P0.05或P0.01);山羊钼镉联合胁迫导致肾线粒体抗氧化功能降低、自由基含量升高,导致细胞促凋亡基因表达量上升,抗凋亡基因的表达量下降,提示钼镉联合胁迫对山羊肾脏有明显损伤。     

Virus Location and Lymphocyte Apoptosis in Lymph Nodes of Piglets Infected with PCV-2

The present study has been performed to understand the location of the virus, type of apoptotic cells, and their relation to lymph nodes of piglets infected with porcine circovirus type Ⅱ （PCV-2）. Nine 32-day-old conventional piglets free of infection with PCV-2 were used, and distributed into three groups： control group （n = 3）, piglets inoculated with PCV-2 alone （PCV-2, n = 3）, and PCV-2 inoculated and KLH immunostimulated group （PCV-2 ＋ KLH, n = 3）. Superficial inguinal lymph nodes from all piglets were collected for histological examination after 32 days postinoculation, and immunohistochemistry for PCV-2 detection. Location of apoptotic cells was detected with TdT-mediated dUTP nick end labeling （TUNEL） and cell cycle, and the apoptotic rates were measured by flow cytometry. The characteristic histopathological lesions of the piglets in PCV-2 and PCV-2 ＋ KLH were lymphocyte depletions in the cortex and paracortex of the lymph nodes, epithelioid-like macrophage infiltration, and intracytoplasmic inclusion bodies presented in epithelioid-like macrophages. PCV-2 was mainly found in epithelioid-like macrophages by immunohistochemistry. In the lymph nodes, lymphocytes presented higher apoptotic rates in the cortex by TUNEL, special B-cell areas, and similar apoptotic cells were found in this compartment in the control. The apoptotic rates of the lymph nodes were 0.41, 3.34, and 4.88% in the control, PCV-2, and PCV-2 ＋ KLH groups by flow cytometry, respectively. The apoptotic rates of lymph nodes for PCV-2 and PCV-2 ＋ KLH piglets were significantly higher than those for the control group （P〈0.05 and P〈0.01）. The proliferation index （PI） was 0.17_＋0.01, 0.12_＋0.01 and 0.12_＋0.04 in the control, PCV-2, and PCV-2 ＋ KLH group, the PI of the control group was higher than that of the other groups, but without the statistical difference. PCV-2 can induce lymphocyte depletion in lymph nodes of piglets by blocking cell proliferation and promoting apoptosis. This is one o