Mdivi-1, mitochondrial fission inhibitor,impairs developmental competence and mitochondrial function of embryos and cells in pigs

Mitochondria are highly dynamic organelles that undergo constant fusion/fission as well as activities orchestrated by large dynamin-related GTPases. These dynamic mitochondrial processes influence mitochondrial morphology, size and function. Therefore, this study was conducted to evaluate the effects of mitochondrial fission inhibitor, mdivi-1, on developmental competence and mitochondrial function of porcine embryos and primary cells. Presumptive porcine embryos were cultured in PZM-3 medium supplemented with mdivi-1 (0, 10 and 50 μM) for 6 days. Porcine fibroblast cells were cultured in growth medium with mdivi-1 (0 and 50 μM) for 2 days. Our results showed that the rate of blastocyst production and cell growth in the mdivi-1 (50 μM) treated group was lower than that of the control group (P < 0.05). Moreover, loss of mitochondrial membrane potential in the mdivi-1 (50 μM) treated group was increased relative to the control group (P < 0.05). Subsequent evaluation revealed that the intracellular levels of reactive oxygen species (ROS) and the apoptotic index were increased by mdivi-1 (50 μM) treatment (P < 0.05). Finally, the expression of mitochondrial fission-related protein (Drp 1) was lower in the embryos and cells in the mdivi-1-treated group than the control group. Taken together, these results indicate that mdivi-1 treatment may inhibit developmental competence and mitochondrial function in porcine embryos and primary cells.     

BHBA对HT22细胞中ACE表达的影响

生酮饮食(KD)已被证实具有神经保护作用,但其机制尚不明确。本试验以小鼠海马神经元HT22为细胞模型,利用2 mmol/Lβ-羟基丁酸酯(BHBA)对HT22细胞进行处理,通过荧光定量PCR,免疫蛋白印迹和信号通路阻断试验,观察BHBA对HT22细胞ACE表达的影响,并探讨参与调控的信号通路。结果表明,2 mmol/L BHBA可通过激活HT22细胞内GPR109A受体和ERK1/2信号通路进而增加HT22细胞中ACE的表达。     

SCINTIGRAPHIC TRACKING OF MESENCHYMAL STEM CELLS AFTER PORTAL,SYSTEMIC INTRAVENOUS AND SPLENIC ADMINISTRATION IN HEALTHY BEAGLE DOGS

Mesenchymal stem cells have been proposed to treat liver disease in the dog. The objective of this study was to compare portal, systemic intravenous and splenic injections for administration of mesenchymal stem cells to target the liver in healthy beagle dogs. Four healthy beagle dogs were included in the study. Each dog received mesenchymal stem cells via all three delivery methods in randomized order, 1 week apart. Ten million fat‐derived allogeneic mesenchymal stem cells labeled with Technetium‐99m (99mTc)‐hexamethyl‐propylene amine oxime(HMPAO) were used for each injection. Right lateral, left lateral, ventral, and dorsal scintigraphic images were obtained with a gamma camera equipped with a low‐energy all‐purpose collimator immediately after injection and 1, 6, and 24 h later. Mesenchymal stem cells distribution was assessed subjectively using all four views. Pulmonary, hepatic, and splenic uptake was quantified from the right lateral view, at each time point. Portal injection resulted in diffuse homogeneous high uptake through the liver, whereas the systemic intravenous injection led to mesenchymal stem cell trapping in the lungs. After splenic injection, mild splenic retention and high homogeneous diffuse hepatic uptake were observed. Systemic injection of mesenchymal stem cells may not be a desirable technique for liver therapy due to pulmonary trapping. Splenic injection represents a good alternative to portal injection. Scintigraphic tracking with 99mTc‐HMPAO is a valuable technique for assessing mesenchymal stem cells distribution and quantification shortly after administration. Data obtained at 24 h should be interpreted cautiously due to suboptimal labeling persistence.     

培养液中铜水平对PK15细胞内铜离子浓度与CuZn-SOD活性的影响

在 PK15细胞无血清液体培养基中添加 0、7.8、15 .6、31.2、6 2 .5 μmol/L Cu,观察了培养液中不同铜水平对PK15细胞内铜离子浓度和 Cu Zn- SOD活性的影响及二者的相关性。结果表明 ,PK15细胞内铜离子浓度及 Cu Zn-SOD活性均随培养液中添加铜离子浓度升高和培养时间的延长而有不同程度的升高 ,其中以添加 Cu31.2 μmol/L组最高 (P<0 .0 5 )。PK15细胞内 Cu Zn- SOD活性与细胞内铜离子浓度和培养时间呈强正相关 (培养时间 :r=0 .873～0 .980 ,P<0 .0 5 ;铜离子浓度 :r=0 .90 0～ 0 .972 ,P<0 .0 5 )     

Exfoliation rate of mammary epithelial cells in milk on bovine mastitis caused by Staphylococcus aureus is associated with bacterial load

The exfoliation rate of mammary epithelial cells (MECs) in milk is affected by physiological, breeding and environmental factors. Little is known about the relationship between the MEC exfoliation into milk and mammary‐infected Staphylococcus aureus (S. aureus) load on bovine mastitis caused by S. aureus. The aim of this study was to investigate the relationship between S. aureus load and the proportion of MEC exfoliation in milk using five substantial bovine mastitis models. In 64 randomly extracted milk samples from udders at 3–21 days after S. aureus infusion, there were various samples with different numbers of S. aureus counts and somatic cell counts. No significant correlations were found between the S. aureus counts and somatic cell count (r = 0.338). In contrast, a significant correlation was noted between S. aureus counts and the proportion of cytokeratin‐positive cells in the milk from the infused udders (r = 0.734, P < 0.01). In conclusion, the increasing MEC exfoliation rate in milk from mastitis udders caused by S. aureus may contribute to reduced milk yield.     

副猪格拉瑟菌5型细胞致死性膨胀毒素CdtB破坏猪呼吸道上皮屏障的机制

旨在探究副猪格拉瑟菌(Glaesserella parasuis,GPS)突破猪呼吸道上皮屏障引起系统性感染的机制。通过超速离心和密度梯度离心提取副猪格拉瑟菌外膜囊泡(outer membrane vesicles, OMVs),OMVs经SDS-PAGE显示,蛋白质条带分布在55～100 ku,经透射电子显微镜(TEM)观察,OMVs的粒径在100～200 nm,纳米粒子直径分析(NTA)结果显示,样品在100～200 nm处的粒子数目最多。进而用制备的HbpA及OmpP2多克隆抗体对OMVs及不含OMVs的细菌上清进行Western blot验证,证实所提取的样品为外膜囊泡,且进一步结果证明细胞致死性膨胀毒素(CDT)在GPS培养物中主要以OMVs的形式存在。用OMVs或CdtB处理猪气管上皮细胞(swine tracheal epithelial cells, STEC)36 h,检测STEC中cleaved-caspase3、ZO-1和Occludin的蛋白表达水平,并用FITC-葡聚糖(FD-4)检测STEC单层细胞的细胞旁通透性。结果发现,OMVs与CdtB处理后凋亡相关蛋...     

敲低组蛋白甲基转移酶ASH1L基因对牛卵丘细胞表达谱的影响

旨在研究干扰类缺失的、小的、同源异形1(absent,small,or homeotic 1-like,ASH1L)甲基转移酶基因表达对牛卵丘细胞表达谱的影响.本研究采用经过筛选的siRNA干扰牛卵丘细胞中ASH1L基因的表达,分别收集干扰组和对照组细胞(野生型)进行RNA测序及基因差异表达、基因功能、信号通路、可变剪...     

细胞自噬对动物卵泡发育调控的研究进展

卵泡是雌性哺乳动物发挥其繁殖能力的基础,其发育是一个动态的过程,主要涉及原始卵泡的形成、卵泡的募集、优势卵泡的选择、成熟卵泡的排卵以及排卵后卵泡的黄体化。卵泡发育的整个过程受内分泌系统、细胞自噬、细胞凋亡等的调控。自噬是一种进化上保守的应激反应过程,通过将细胞内物质包裹形成自噬体并传递到溶酶体中进行降解,以帮助细胞维持胞内物质代谢平衡,其在卵泡发育的过程中发挥着重要作用,一方面它能够通过降解或回收受损的蛋白质或有害代谢产物缓解应激造成的卵泡损伤,另一方面它又通过产生大量自噬体导致细胞器过度降解而引起卵泡闭锁。自噬对卵泡发育的调控需要PI3K-Akt-mTOR、MAPK-ULK1、ERK1/2、Sirt1-FOXO1-Atg7等多种经典信号通路的参与,这些信号通路在激素、氧化应激、细胞饥饿等的刺激下,通过独立作用或相互作用促进或抑制自噬调控卵泡细胞的生理活动。目前已知不同的自噬水平对卵泡细胞的存活具有不同作用,但关于决定细胞能否存活的自噬水平的研究还比较少。此外,自噬对卵泡发育调控的研究主要集中在颗粒细胞中,而对卵母细胞的成熟和卵泡膜细胞的作用的报道较少。文章简述了自噬在卵巢储备的形成、...