自噬通量在小檗碱抑制奶牛子宫内膜上皮细胞凋亡中的作用

为探究小檗碱对脂多糖诱导奶牛子宫内膜上皮细胞（BEECs）凋亡的调控机理,首先利用CCK8法确定添加药物的细胞毒性,然后将BEECs分为空白（C）组、脂多糖（LPS）组（1μg/mL）、小檗碱（BBR）组（BBR10μmol/L+LPS 1μg/mL）、氯喹（CQ）组（CQ 20μmol/L+BBR 10μmol/L+LPS 1μg/mL）,用流式细胞术检测细胞凋亡率,用Western blot检测BAX、BCL2、LC3II、P62蛋白表达量。结果显示：LPS组BEECs凋亡率和BAX/BCL2较空白组升高（P<0.01）;BBR组细胞凋亡率和BAX/BCL2低于LPS组（P<0.01）;CQ组细胞凋亡率高于其他3组（P<0.01）,而BAX/BCL2与空白组差异不显著。LPS组BEECs表达P62高于空白组（P<0.01）;BBR组与LPS组相比LC3II升高（P<0.05）、P62降低（P<0.01）;CQ组LC3II高于其他3组（P<0.01）,CQ组P62与空白组、BBR组差异显著而与LPS组差异不显著。说明LPS降低BEECs自噬通...     

小麦饲料中添加木聚糖酶对高原型藏羊胃肠道发育的影响

旨在分析小麦粉中添加木聚糖酶(xylanase, Xyl)对藏羊羔羊胃肠道发育的影响。选择同质性良好的2～3月龄高原型藏羊公羔60只,随机分为两组,每组30只,各组内设置5个重复。对照组(Control group)饲喂不含木聚糖酶的日粮,试验组(Test group)饲喂含0.2%木聚糖酶的日粮,试验分为预饲期10 d和正试期90 d。饲养试验结束时,两组各随机选取5只羔羊进行屠宰,采集消化道组织置于多聚甲醛固定,并收集血液、瘤胃和空肠内容物。结果显示：1)与对照组相比,试验组瘤胃的角质层厚度、颗粒层厚度,瓣胃的乳头长度、乳头宽度、角质层厚度、颗粒层厚度,皱胃的黏膜厚度以及十二指肠的肌层厚度,空肠的绒毛高度、黏膜厚度,回肠的肌层厚度显著提高(P<0.05);2)对照组瘤胃和血清的LPS含量显著高于试验组(P<0.05);3)相较于对照组,试验组瘤胃的糜蛋白酶、胰蛋白酶、脂肪酶活性以及空肠的糜蛋白酶、α-淀粉酶活性显著提高(P<0.05);4)试验组瘤胃的总抗氧化能力水平、谷胱甘肽过氧化物酶及过氧化氢酶活性以及空肠的总抗氧化能力水平、谷胱甘肽过氧化物酶、超氧化物歧化酶...     

棕榈粕替代部分玉米对藏羊母羊小肠形态发育、消化酶活性及抗氧化功能的影响

试验旨在探究精料补充料中使用不同比例棕榈粕替代玉米对藏羊母羊肠道组织形态学、消化酶活性、pH、脂多糖以及抗氧化能力的影响。选取初始体重相近且健康的2～3月龄高原型藏羊母羊120只,随机分为4组,每组30只,每组6个重复,每个重复5只母羊,分别饲喂0%、15%、18%和21%水平的棕榈粕替代精料中玉米。试验期97d。结果显示：1)0%组与15%组间小肠各段的绒毛高度、绒毛宽度、隐窝深度、黏膜厚度以及绒毛高度/隐窝深度差异均不显著（P>0.05）;2)0%组空肠的α-淀粉酶、纤维素酶、脂肪酶及糜蛋白酶显著或极显著小于15%组（P<0.05或P<0.01）;3)0%组空肠的谷胱甘肽过氧化物酶、过氧化氢酶活性显著低于15%组与18%组（P<0.05）,相较于21%组差异不显著（P>0.05）;4)18%组与21%组空肠的脂多糖含量显著或极显著高于0%组（P<0.05或P<0.01）,与15%组相比差异不显著（P>0.05）。由此可见,棕榈粕可替代部分玉米饲喂藏羊母羊,推荐替代玉米的比例为15%。     

活性氧参与脂多糖诱导小鼠巨噬细胞RAW 264.7活化诱导凋亡

研究活性氧(ROS)与脂多糖(LPS)诱导后巨噬细胞活化诱导凋亡的关系。体外培养小鼠巨噬细胞RAW264.7,采用不同浓度LPS诱导细胞,添加100μmol/L H2O2增加细胞内ROS,或用姜黄素(7.5μmol/L)降低细胞内ROS;流式细胞术分析细胞凋亡、线粒体膜电位(MMP)和ROS。结果显示,RAW264.7细胞凋亡和细胞内ROS水平均随LPS浓度增加而增加,高浓度的LPS导致MMP下降;与LPS(8μg/m L)单独作用组比,H2O2增加ROS,并导致凋亡细胞百分率增加;姜黄素降低LPS诱导的细胞凋亡,同时减少细胞内ROS。表明活性氧参与LPS诱导小鼠巨噬细胞RAW264.7活化诱导凋亡。     

黄花蒿乙醇提取物对脂多糖诱导损伤奶牛乳腺细胞中乳蛋白合成的影响

本试验旨在通过奶牛乳腺上皮细胞(BMECs)体外培养技术,研究黄花蒿乙醇提取物(AAE)对脂多糖(LPS)诱导损伤BMECs活力以及乳蛋白合成相关基因表达的影响。试验分为5组,对照组:BMECs用基础培养基常规培养15 h;模型组:BMECs用基础培养基常规培养3 h+10μg/mL LPS处理12 h;AAE+LPS组:BMECs分别用含3、6、12μg/mL AAE的基础培养基培养3 h+10μg/mL LPS处理12 h。结果表明:1)与对照组相比,模型组的BMECs活力显著降低(P0.05)。与模型组相比,6和12μg/mL AAE组的BMECs活力显著增加(P0.05)。2)与模型组相比,12μg/mL AAE组总蛋白含量显著增加(P0.05),6、12μg/mL AAE组β-酪蛋白含量显著增加(P0.05),3、6、12μg/mL AAE组αs1-酪蛋白含量显著增加(P0.05)。3)与模型组相比,12μg/mL AAE组α-酪蛋白(CSN1S1)基因表达量显著增加(P0.05),3、6、12μg/mL AAE组β-酪蛋白(CSN2)、哺乳动物雷帕霉素靶蛋白(mTOR)、信号转导和转录激活因子5(STAT5)、真核细胞翻译启动因子4E结合蛋白1(EIF4EBP1)、核糖体S6蛋白激酶1(S6K1)、Janus激酶2(JAK2)基因表达量显著增加(P 0.05),6、12μg/mL AAE组真核起始因子4E(EIF4E)基因表达量显著增加(P0.05)。由此可见,12μg/mL AAE对LPS诱导损伤BMECs活性和乳蛋白合成有较好的预保护效果。     

Extraction,Purification and Activity Analysis of Lipopolysaccharides from Escherichia coli Isolated from Swine

In order to obtain higher purity and high yield of LPS isolated from Escherichia coli, and evaluate its virulence,excrement of diarrhea piglet was collected as pathological samples, of which, some strains of Escherichia coli were isolated and identified. And furthermore, a strain of Escherichia coli was acquired by 16S rDNA identification. The LPS from this strain of Escherichia coli was isolated with the hot-phenol water method and purified by DNaseⅠ, RNaseA, proteinase K treatment and alcohol precipitation. The polysaccharide, protein and nucleic acid content of purified LPS were detected by phenol-sulfuric acid method, Bradford method and UV points spectrophotometric method. And its bioactivity and acute median lethal dose was determinated by tachypleus amebocyte lysate (TAL) method and acute toxicity experiment method, respectively. The results showed that a strain of highly pathogenic Escherichia coli was successfully isolated. The average yield of purified LPS was 3.74%, the proportion of the polysaccharide was 13.40%, the protein was 0.48%, and the nucleic acid was 0.06%. SDS-PAGE electrophoresis and silver stain showed that the bands were mainly concentrated in the range of 14 to 160 ku. The LAL bioactivity of the prepared LPS was 1.21×10⁷ EU/mg, LD₅₀ of the mouse abdominal cavity was 31.86 mg/kg. The results revealed that the diarrhea piglets in the farm was caused by pathogenic Escherichia coli,purified LPS from this strain of Escherichia coli had the advantages of higher purity, high yield and strong virulence, which would provide theoretical data for clinical prevention and treatment of diarrhea piglets.