围产期酮病牛脂联素mRNA和甘油三酯脂肪酶mRNA的表达

选择自然发生酮病牛10头、正常对照组奶牛（健康牛）10头。分别采取酮病牛和产后不同时期的奶牛尾部皮下脂肪组织,应用荧光定量PCR法检测脂联素（ADPN）mRNA与甘油三酯脂肪酶（HSL）mRNA表达的状况。酮病牛与对照组（健康）牛分别颈静脉采血,测定葡萄糖（GLU）、游离脂肪酸（NEFA）、β羟丁酸（BHBA）、甘油三酯（TG）以及胰岛素（INS）、胰高血糖素（GN）等相关指标。结果表明：酮病奶牛血糖显著降低,血浆NEFA和BHBA、血清INS降低和INS/GN变小;酮病奶牛HSLmRNA的表达低于围产期健康奶牛产后14d和产后28d,但ADPN mRNA却极显著地高于围产期健康奶牛产后的各生理时期,表明酮病牛因能量代谢紊乱脂肪动员产物NEFA和BHBA的增多,激发导致ADPN mRNA的表达,但ADPN mRNA的表达增加到一定量时反而会降低HSL mRNA的表达,说明ADPN参与了脂肪代谢的调节。     

地菍总黄酮对胰岛素抵抗小鼠的干预作用及机制研究

本研究旨在探讨地菍总黄酮（TFMD）对胰岛素抵抗（IR）小鼠的干预作用及其机制。试验以灌胃高脂乳剂建立胰岛素抵抗小鼠模型，同时灌胃给予地菍总黄酮混悬液（高剂量（600 mg/kg）、中剂量（300 mg/kg）及低剂量（150 mg/kg））进行治疗，治疗周期为30 d。试验结束后测定小鼠血清中空腹血清胰岛素（FINS）、空腹血糖（FBG）、胰岛素抵抗指数（HOMA-IR）、胰岛素敏感指数（ISI）、血清甘油三酯（TG）、总胆固醇（TC）、低密度脂蛋白（LDL）及高密度脂蛋白（HDL）水平；利用实时荧光定量PCR法检测各组小鼠肝脏中胰岛素受体（InsR）、过氧化物酶体增殖物激活受体-γ（PPAR-γ）及骨骼肌中葡萄糖转运体4（GLUT4）的mRNA表达水平；利用免疫组化技术检测各组小鼠肝脏中InsR、PPAR-γ和骨骼肌中GLUT4的蛋白表达水平。结果显示，与模型组相比，地菍总黄酮能降低IR小鼠的体重，降低血清FBG、FINS及HOMA-IR水平，升高ISI水平（P<0.01，P<0.05）；降低血清TG、TC及LDL含量，升高HDL含量（P<0.01，P<0.05）；同时提高IR小鼠肝脏中InsR、PPAR-γ及骨骼肌中GLUT4的mRNA表达量，提高小鼠肝脏InsR、PPAR-γ及骨骼肌中GLUT4的蛋白表达水平（P<0.01，P<0.05）。以上试验结果证实，地菍总黄酮能有效缓解小鼠试验性胰岛素抵抗症状，该活性与调节糖脂代谢、增强胰岛素敏感性有关。     

"酮病制剂1号"防治奶牛酮病效果观察

选取高产奶牛20头，根据乳酮阳性，血浆β-羟丁酸和血糖浓度分为健康牛，亚临床酮病牛和临床型酮病牛。将试验奶牛分4组，每组5头，Ⅰ组为亚临床酮病给药组，Ⅱ组为临床酮病给药组，Ⅲ组为酮病未给药对照组，Ⅳ组为健康未给药对照组。再给“酮病制剂1号”前后测定各组奶牛泌乳量、血浆β-羟丁酸及血糖指标。结果表明：“酮病制剂1号”使产后酮病奶牛泌乳量增加不显著（P〉0．05）；给药后，亚临床酮病Ⅰ组与临床酮病Ⅱ组奶牛血浆β-羟丁酸显著降低（P〈0.01），血糖浓度显著增加（P〈0．01）；未给药酮病Ⅲ组和对照Ⅳ组奶牛血浆β-羟丁酸和血糖给药前后差异不显著。结论：“酮病制剂1号”通过降酮升糖的途径发挥防治奶牛酮病的作用。     

围生期能量负平衡奶牛胰岛素抵抗的研究进展

围生期奶牛能量负平衡可引起脂代谢紊乱并常伴有胰岛素抵抗,而胰岛素抵抗又进一步促进了脂代谢紊乱和能量负平衡。胰岛素抵抗是奶牛能量代谢性疾病酮病和脂肪肝的病理学基础,胰岛素抵抗奶牛主要临床特征是高非酯化脂肪酸血症、高酮血症和低胰岛素敏感性指数。高产奶牛易发生能量代谢性疾病,造成巨大经济损失。炎症、内质网应激和线粒体功能紊乱参与了胰岛素抵抗的发生。文章综述了奶牛胰岛素抵抗与能量代谢障碍性疾病之间的研究进展。     

胰岛素受体mRNA在新生犊牛组织中的表达

应用半定量RT-PCR方法检测了新生犊牛中枢神经系统和外周组织中胰岛素受体（insulin receptor，InsR）基因的表达。结果表明，InsR基因在肝、皮下脂肪、半腱肌、胰、肾皮质、脾、心、肺、下丘脑、肠系膜淋巴结、主动脉、十二指肠、结肠、垂体、大脑皮质、小脑皮质中都有表达。其中，肝、半腱肌、下丘脑、胰、主动脉、垂体中InsR基因的表达量显著多于其他组织（P〈0．05）。InsR基因在各组织中的广泛分布表明胰岛素在体内具有广泛的生理功能。     

不同能量摄入对围产期奶牛脂肪组织胰岛素受体mRNA丰度的影响

将年龄、胎次相同,年产奶量大于5000kg的围产期健康奶牛30头随机分为3组,每组10头。按照中国奶牛饲养标准(2000),低能组饲喂减少20%日粮(能量摄入80%组),对照组饲喂标准日粮(能量摄入100%组),高能组饲喂增加20%日粮(能量摄入120%组),试验从产前14天开始至产后28天结束,产后各组奶牛均饲喂标准日粮。采用半定量RTPCR方法检测了不同能量摄入对围产期奶牛脂肪组织胰岛素受体(InsR)mRNA丰度的影响。结果表明:各组脂肪InsRmRNA相对表达量从产前14天至产后28天均呈现先升高后降低的趋势,产后14天达到最大值,且产后高于产前;低能组产前InsRmRNA相对表达量低于高能组和正常组,产后1天至产后28天高于高能组和正常组,产后14天显著高于高能组(P<0.01)。可见,干乳期低能饲喂奶牛,产后脂肪InsRmRNA表达增加,提示产前适当降低能量摄入水平,有利于胰岛素抑制泌乳初期脂肪动员作用的发挥。     

奶牛酮病的发病原因及防治措施

奶牛酮病（ketosis），又称奶牛酮血症，是奶牛最常见的糖和脂肪代谢紊乱疾病。临床上以低血糖、高血脂、酮血、酮尿、脂肪肝、酸中毒及体蛋白消耗和食欲减退或废绝为特征。     

围产期奶牛NEFA的营养调控研究进展

非酯化脂肪酸（NEFA）升高是围产期奶牛能量负平衡的标志，也是导致奶牛发生脂肪肝和酮病的直接原因。在过去的30余年里人们对围产期奶牛NEFA营养调控的方法和机制进行了大量研究，文章就该方面的研究进展作一综述。     

奶牛ACC基因SYBRGreenⅠ荧光定量PCR检测方法的建立

乙酰辅酶A羧化酶(ACC)在奶牛肝脏中脂肪酸的合成及氧化代谢过程中起着关键作用,ACC mRNA的表达水平对于研究奶牛酮病、脂肪肝等物质代谢障碍性疾病具有重要意义。研究采用双标准曲线法,成功建立了奶牛ACC基因荧光定量检测方法。结果表明,建立的SYBR Green I荧光定量PCR方法具有操作简便、特异性强、定量准确等优点,为进一步对ACC基因定量分析,研究其在奶牛酮病、脂肪肝等营养代谢性疾病中的作用奠定了基础。     

不同能量摄入水平对围产期奶牛肝载脂蛋白E mRNA丰度的影响

将年龄、胎次相同，年产奶量大于5000kg的围产期健康奶牛30头随机分为3组，每组10头。低能组饲喂中国奶牛饲养标准（2000）减少20％日粮（能量摄入80％组）、对照组饲喂中国奶牛饲养标准（2000）日粮（能量摄入100％组）和高能组饲喂中国奶牛饲养标准（2000）增加20％日粮（能量摄入120％组），试验从产前28d开始至产后56d结束，产后各组奶牛均饲喂标准日粮。采用内对照RT-PCR方法检测了不同能量摄入围产期奶牛肝组织载脂蛋白E（apolipoprotein E，apoE）mRNA丰度。结果表明：apoE mRNA相对表达量从产前28d至产后56d各组均呈现先升高后降低的趋势，各组apoE mRNA水平均在产后一天达到最大值（P〈0．01），且产后均高于产前（产后56d除外）；产前14d至产后28d低能组apoE mRNA相对表达量均高于其他两组（P〈0．05或P〈0．01），高能组在产后28d和56d高于对照组（P〈0．01）。可见，围产期奶牛能量摄入水平对肝组织载脂蛋白E mRNA丰度有显著影响。     

Coordinated gene expression in adipose tissue and liver differs between cows with high or low NEFA concentrations in early lactation

Dairy cows with high and low plasma non-esterified fatty acid (NEFA) concentrations in early lactation were compared for plasma parameters and mRNA expression of genes in liver and subcutaneous adipose tissue. The study involved 16 multiparous dairy cows with a plasma NEFA concentration of >500 μmol/l [n = 8, high NEFA (HNEFA)] and <140 μmol/l [n = 8, low NEFA (LNEFA)] in the first week post-partum (pp). Blood samples, adipose and liver tissues were collected on day 1 (+1d) and at week 3 pp (+3wk). Blood plasma was assayed for concentrations of metabolites and hormones. Subcutaneous adipose and liver tissues were analysed for mRNA abundance by real-time qRT-PCR encoding parameters related to lipid metabolism. Results showed that mean daily milk yield and milk fat quantity were higher in HNEFA than in LNEFA cows (p < 0.01), and the NEB was more negative in HNEFA than in LNEFA in +3wk too (p < 0.05). HNEFA cows had slightly lower (p < 0.1) insulin concentrations than LNEFA cows across the study period, and the body condition score decreased more from +1d to +3wk in HNEFA than in LNEFA (p = 0.09). The mRNA abundance of genes in the liver related to fatty acid oxidation (carnitine palmitoyltransferase 2 and very long chain acyl-coenzyme A dehydrogenase) and ketogenesis (3-hydroxy-3-methylglutaryl-coenzyme A synthase 2) were lower in HNEFA than in LNEFA cows. No differences between the two groups were observed for mRNA expression of genes in adipose tissue. The number of calculated significant correlation coefficients (moderately strong) between parameters in the liver and in adipose tissue was nearly similar on +1d, and higher for HNEFA compared with LNEFA cows in +3wk. In conclusion, dairy cows with high compared with low plasma NEFA concentrations in early lactation show differentially synchronized mRNA expression of genes in adipose tissue and liver in +3wk that suggests a different orchestrated homeorhetic regulation of lipid metabolism.     

神经内分泌因子对体外培养新生犊牛肝细胞MTP mRNA表达的影响

为了阐明神经内分泌因子在奶牛肝脂蛋白组装与转运过程中的调控作用,通过向体外培养的新生犊牛肝细胞添加胰岛素、胰高血糖素和瘦蛋白,采用内对照RT-PCR方法检测神经内分泌因子对体外培养新生犊牛肝细胞微粒体甘油三酯转运蛋白(MTP)mRNA丰度的影响。结果显示,随着细胞培养液中胰岛素和胰高血糖素浓度升高,MTP mRNA的丰度呈现剂量依赖性下降(P<0.01);随着瘦蛋白浓度的增加,MTP mRNA的丰度先升高后降低(P<0.01或P<0.05)。这表明胰岛素和胰高血糖素对肝细胞MTP mRNA表达具有抑制作用,呈现剂量依赖性;瘦蛋白对肝细胞MTP mRNA表达具有低剂量促进而高剂量抑制的双重作用,且均呈现剂量依赖性。     

Plasma lipids in normal cows around partus and in cows with metabolic disorders with and without fatty liver

Free fatty acids, cholesterol and phospholipids in plasma were studied from 6 weeks before to 6 weeks after calving in 16 normal multiparous cows. The same plasma lipids were studied the day after calving in 20 normal primiparous cows. Ten of these were fed according to standards and the other 10 were overfed the last 3 weeks prepartum. The plasma lipids were also analysed in 16 cows with left displacement of the abomasum and fatty liver, and in 16 cows with ketosis with no or only slight fat infiltration of the liver. In the normal cows there was a rise in FFA-level and a reduction in cholesterol and phospholipids from 6 weeks before to the day after calving. Thereafter there was a reduction of FFA-level and rise in cholesterol and phospholipids. Increased feed intensity had no effect on plasma lipids at calving. The level of the plasma lipids in cows with fatty liver differed very much from the amounts in normal cows at corresponding time from calving. Cows with ketosis had high FFA-level but the amount of cholesterol and phospholipids differed very little from normal cows.     

The effects of a single injection of dexamethasone-21-isonicotinate on the lymphocyte functions of dairy cows at two weeks post partum

Dexamethasone is a potent therapeutic for treatment of the fatty liver syndrome or primary ketosis in post partum dairy cows. Reservations exist, however, among practitioners with respect to the risk of immunosuppression induced by corticosteroids. The aim of this study was to investigate the effect of a single injection of dexamethasone-21-isonicotinate on distinct immune functions of postpartum dairy cows because only scarce information is available on the effects of corticosteroid preparations when administered at a dosage and frequency for treatment of the fatty liver syndrome or primary ketosis. Sixteen Swedish red-pied dairy cows, between days 9 and 15 post partum, were allotted to either a control group (n = 8) or a treatment group (n = 8). The cows in the treatment group received a single intramuscular injection of a dexamethasone-21-isonicotinate suspension at a dosage of 0.02 mg/kg i.m. at the start of the experiment. White blood cell counts and selected lymphocyte functions (lymphocyte proliferation, expression of lymphocyte markers and the b2 and a4 chain of adhesion molecules belonging to the integrin family) and some parameters of the energy metabolism (glucose, insulin) were determined before the administration of corticosteroids (day 0) and subsequently at days 2, 4, 7 and 9 of the experiment. Changes in glucose and insulin were within the target range for treatment of the fatty liver syndrome or primary ketosis. Significant (P < 0.05) increases in the number of circulating white blood cells were observed in treated cows on the second day following treatment which was exclusively caused by an increase in the number of circulating neutrophils. Lymphocyte blastogenesis in response to ConA and the percentages of lymphocytes positive for CD2, CD4, CD8, CD49d and CD18 as well as the intensity of CD49d expression did not differ between the treatment and control groups. There was, however, a significant reduction (P < 0.01) in the intensity of CD18 expression on lymphocytes in the treated animals on the fourth day after treatment. In conclusion, a single administration of dexamethasone-21-isonicotinate in a dosage of 0.02 mg/kg i.m. at two weeks post partum in healthy cows had a significant but highly transient effect on CD18 expression on lymphocytes and the number of peripheral blood neutrophils, but did not affect lymphocyte blastogenesis or lymphocyte subpopulation patterns in peripheral blood.     

Study on the Intervention Effect and Mechanism on Insulin Resistant Mice of Total Flavone from Melastoma dodecandrum Lour.

This experiment was conducted to explore the intervention effect and mechanism on insulin resistance (IR) mice of total flavonoids from Melastoma dodecandrum Lour.(TFMD).The model of IR mice was established by intragastric administration of high-fat emulsion,while 600,300 and 150 mg/kg TFMD were administered once daily for 30 days.After the end of the experiment the mices' fasting blood glucose (FBG),fasting serum insulin (FINS),serum total cholesterol(TC),triglyceride (TG),high density lipoprotein(HDL) were determined.The mRNA expression level of liver insulin receptor(InsR),fat peroxisome proliferator-activated receptor-γ(PPAR-γ),and glucose transporter 4 gene (GLUT4) in skeletal muscle were detected by Real-time quantative PCR,and the protein levels were detected by immunohistoche mical method.The results showed that TDMF could reduce the body weight of IR mice,decrease the level of serum FBG,FINS and HOMA-IR,increase the level of ISI,decrease the content of serum TG,TC and LDL,and increase the content of HDL(P<0.01,P<0.05);And the mRNA expression of INSR,PPAR-γ in liver and GLUT4 in skeletal muscle of IR mice were increased(P<0.01,P<0.05),and the protein expression level of InsR,PPAR-γ in liver and GLUT4 in skeletal muscle of IR mice were increased(P<0.01,P<0.05).The results confirmed that the TFMD could relieve experimental insulin resistance in mice,and the activity was related to the regulation of glucose and lipid metabolism and the enhancement of insulin sensitivity.     

胰岛素、胰高血糖素对体外培养脂肪细胞胰高血糖素受体mRNA丰度的影响

为阐明胰岛素、胰高血糖素在奶牛脂肪代谢中的调控作用,应用RT-PCR法观察了胰岛素、胰高血糖素对体外培养脂肪细胞胰高血糖素受体(GLNR)mRNA丰度的影响。结果发现,随着培养液中胰岛素质量浓度的升高,GLNR mRNA表达逐渐增加(P<0.05);而随着培养液中胰高血糖素浓度的升高,GLNR mRNA表达逐渐降低(P<0.01)。本研究结果表明,胰岛素、胰高血糖素直接调控奶牛脂肪细胞胰高血糖素受体mRNA的表达。     

Comparative proteomic analysis of livers from ketotic cows

The aim of this study was to examine molecular pathways activated in the liver of the ketotic cow to gain further understanding of the pathogenic effects of ketosis. We used two-dimensional-electrophoresis (2D-E) and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF-TOF) tandem mass spectrometry to compare the liver proteomic profile between ketotic and normal cows. Thirty-eight different proteins were identified between these groups by 2D-E gel. These proteins play a role in energy metabolism, carbohydrate degradation, fatty acid metabolism, amino acid metabolism, antioxidation, cell structure, nucleotide metabolism, and protein metabolism. This is the first time the majority of these proteins have been reported in cow ketosis. Possible explanations for the differential expression patterns and their relevance to the pathogenesis of ketosis in the cow are discussed. This study is the first to provide a detailed insight into protein expression patterns in the liver of the ketotic cow, and the different proteins involved in the pathogenesis of bovine ketosis.     

Relationship between ketosis and dairy cows’ blood metabolites in intensive production farms of the periurban area of Dakar

This study which involved 140 Holstein and Montbeliard was carried out in the periurban area of Dakar with the aim to establish the relationship between ketosis, milk production and biochemical blood metabolites. The results showed that ketosis is a real problem in periurban farms around the city of Dakar with high proportions of 33.57% for subclinical ketosis and 6.43% for clinical ketosis. In their second month of milking, cows with subclinical ketosis had a decrease of 12.4 and 15.,6% in milk yield respectively for Montbeliard and Holstein, whereas cows with clinical ketosis had a decrease of 18.6 and 26%. Ketogenic cows (subclinical and clinical) have significantly lower average levels of blood glucose (p<0.05) and significantly higher average levels of blood urea (p<0.05) than cows with normal blood beta-Hydroxy Butyrate (BbHB) levels. Also, from one farm to another, significant difference was recorded with concentration of total proteins and globulin, calcium and magnesium.