Short-chain fatty acids inhibit bovine rumen epithelial cells proliferation via upregulation of cyclin-dependent kinase inhibitors 1A,but not mediated by G protein-coupled receptor 41

Short-chain fatty acids (SCFAs) play a critical role in regulation of rumen epithelial growth. The mechanisms underlying the regulatory effects of SCFAs on the proliferation of bovine rumen epithelial cells (BRECs) remain unknown; however, SCFAs can bind to G protein-coupled receptor 41 (GPR41); hence, the regulatory effects of SCFAs on BRECs proliferation may be mediated by GPR41. Here, we investigated the molecular mechanisms underlying the effects of SCFAs and GPR41 on BRECs proliferation. We demonstrated that SCFAs activate the expression of GPR41 and inhibit (p < .05) BRECs proliferation, while the GPR41 knockdown (GPR41KD) BRECs exhibited (p < .05) slow proliferation compared with controls. The treatment of BRECs with 10 mM SCFAs significantly enhanced (p < .05) expression of cyclin-dependent kinase inhibitors 1A (CDKN1A), 2A (CDKN2A) and 2B (CDKN2B) and inhibited (p < .05) their transition from G1 to S phase of the cell cycle, compared with controls. Remarkably, the GPR41KD BRECs treated with SCFAs restored high level of CDKN1A, relative to GPR41KD BRECs, but did not affect (p > .05) the expression of CDKN2A and CDKN2B. The GPR41KD BRECs had significantly reduced (p < .05) cyclin-dependent kinase 4 (CDK4) and cyclin D2 mRNA abundance compared with controls. The GPR41KD BRECs treated with SCFAs significantly decreased (p < .05) CDK4, cyclin D2, CDKN2A and CDKN2B mRNA abundance compared with BRECs treated with SCFAs. Overall, our results demonstrated that downregulation of CDK4 and cyclin D2 likely mediates the inhibitory effects of GPR41KD on BRECs proliferation. Additionally, CDKN1A plays a vital role in mediating the inhibitory effect of SCFAs on the BRECs proliferation, and that these changes are not mediated by GPR41.     

瘦蛋白(Leptin)对体外培养新生牛脂肪细胞Leptin、HSL基因表达的影响

取单层生长良好的脂肪细胞,采用单因素重复试验,分别添加0、2.5、5、10、50、100μg/L的外源牛重组瘦蛋白(Leptin),培养12h后提取总RNA,每个处理3个重复(孔),应用半定量PCR方法检测外源Leptin对脂肪细胞Leptin mRNA和HSL mRNA水平的影响.结果表明,Leptin对脂肪细胞内Leptin mRNA表达具有抑制作用,但一定浓度的Leptin能提高脂肪细胞内HSL mRNA水平,其中以5μg/L Leptin的作用最为明显;当Leptin的质量浓度超过5μg/L,HSL mRNA表达水平呈下降趋势.     

Localization of leptin and leptin receptor in the bovine adenohypophysis

The present study was carried out to detail the cellular localization of leptin (Lep) and the leptin receptor (LepR) in the bovine adenohypophysis. Lep immunoreactivity (Lep-ir) was found in about 30% of adenohypophysial cells in the gland. Immunochemistry of Lep and specific hormones using serial sections revealed that Lep-ir was present in 60.4% of somatotrophs, 15.9% of gonadotrophs, 6.5% of mammotrophs, 6.5% of thyrotrophs and 2.4% of corticotrophs. Both the common short isoform (OBRa) and the long isoform (OBRb) of LepR mRNA were expressed in the bovine adenohypophysis. LepR immunoreactivity (LepR-ir) was found in only 2.8% of the adenohypophysial cells and over 50% of LepR-ir cells were gonadotrophs, in which most of the cells were distributed in the zona tuberalis. The findings on Lep and LepR in the adenohypophysial cells indicate that Lep may regulate gonadotroph function through autocrine/paracrine pathway in the bovine adenohypophysis.     

多不饱和脂肪酸的营养作用及其基因表达调控

多不饱和脂肪酸(PUFA)是一种重要的营养物质,是所有细胞膜的重要成分,具有重要的生理功能。本文着重论述了PUFA的营养功能和其调控基因表达的分子机制。     

Sunflower-seed oil,rapidly-degradable starch,and adiposity up-regulate leptin gene expression in lactating goats

We conducted experiments to evaluate the effects of lipid supplementation and the nature of starchy concentrate on the regulation of leptin synthesis in lactating goats. Multiparous goats in mid- to late lactation received diets based on different forages and containing plant oil or seeds rich in either 18:1c9, 18:2n-6 or 18:3n-3 corresponding to 3%–7% dry matter (DM) as lipid supplements, or diets based on concentrate as either rapidly or slowly degradable starch. The isoenergetic replacement of a part of the concentrate by either oleic sunflower-seed oil, formaldehyde-treated linseeds, or linseed oil did not modify leptinemia and the leptin mRNA concentration in adipose tissues, suggesting a lack of effect of 18:1c9, 18:3n-3, or their biohydrogenation products. Conversely, leptinemia and the leptin mRNA abundance were increased (by 20% and 140%, respectively, P < 0.05) in goats fed sunflower-seed oil under a grassland hay-based diet but not a maize silage-based diet, at similar energy intakes and adiposity. Thus, 18:2n-6 per se may up-regulate leptin gene expression, but the effect could be blunted by other fatty acids formed during the ruminal digestion of sunflower-seed oil when combined with maize silage. Consumption of rapidly but not slowly degradable starch increased (by 17%, P < 0.05) leptinemia. Moreover, during lactation, plasma leptin was positively correlated (P < 0.05) to adiposity parameters and negatively correlated to fiber intake. The results suggest that leptinemia responds poorly to nutritional factors in lactating goats, thus highlighting the physiological need to sustain hypoleptinemia during lactation.     

脂肪酸对基因表达的影响及调控

脂肪酸是一类重要的营养物质,是动物体内的能量来源,同时也是生物膜的组成成分,在细胞生化过程中也同样起着重要作用。研究发现,脂肪酸可通过对基因表达的影响,对代谢、生长发育以及细胞分化发挥重要的调控作用。真核生物基因表达的调控大致可分为转录前、转录、转录后、翻译和翻译后等5个阶段的调控。脂肪酸通过细胞膜受体信号途径和转录因子活化途径调节基因表达,而多不饱和脂肪酸主要从基因转录和mRNA的稳定性两个方面调节基因表达。文章综述了脂肪酸,尤其是多不饱和脂肪酸对基因表达的影响及调控机制。     

Effect of fatty acids added to the milk replacer on white scour and excretion of fatty acids in Holstein calves

In order to examine the relationship between white scour and fatty acids, we added fatty acids to the milk replacer. Twenty healthy Holstein calves were divided into 4 groups, five calves per group; a control group with no fortified fatty acid, and 3 groups fed either with oleic, stearic, or palmitic acid, respectively. The calves were fed milk replacer (5% of the calf's body weight) twice a day but the fatty acids (2 wt % of milk replacer) were added only once. The fecal and blood samples were obtained at 0, 12, 24, 36, and 48 h after feeding of the acids. All five calves in the palmitic acid group, and 3 out of 5 calves each in the stealic and the oleic acid groups had whitish feces after feeding fatty acid. The stearic acid group had a significantly elevated stearic acid concentration in the feces during 24–36 h compared to the pre-feeding level. The fecal concentration of palmitic acid increased significantly at 24–36 h in the palmitic acid group. We concluded that the long-chain saturated fatty acids are one of the causes of white scour in calves.     

The effect of dietary barley level on volatile fatty acid concentration and manure ammonia emissions in finishing pigs

An experiment was conducted to investigate the linear and quadratic effects of barley on volatile fatty acid (VFA) profile and manure ammonia emissions in pigs. Sixteen boars (66 kg) were assigned to one of four treatments (n = 4): (T1) 0 g/kg barley (control), (T2) 222 g/kg barley, (T3) 442 g/kg barley and (T4) 666 g/kg barley. Barley was substituted for wheat. The diets were formulated to contain similar concentrations of digestible energy and lysine, soya oil was adjusted to maintain net energy. Increasing barley levels caused a linear decrease in colonic (P < 0.01) and caecal pH (P < 0.001). Total caecal VFA and propionic acid were affected by dietary barley level (quadratic, P < 0.05). There was a linear decrease in the proportion of acetic acid (P < 0.001), isobutyric acid (P < 0.01), isovaleric acid (P < 0.05) with increasing levels of barley in both the caecum and colon. There was a linear increase in the proportion of propionic acid (P < 0.001) and butyric acid (P < 0.05) with increasing barley in the colon. Barley based diets had a lower ammonia emission from 96 to 240 h (P < 0.05) and from 0 to 240 h (P < 0.05) than the wheat based diet. In conclusion, a high level of barley is required to reduce offensive branch chained VFAs.     

Dietary fatty acids in gut health: Absorption,metabolism and function

In biological responses, fatty acids (FA) are absorbed and metabolized in the form of substrates for energy production. The molecular structures (number of double bonds and chain length) and composition of dietary FA impact digestion, absorption and metabolism, and the biological roles of FA. Recently, increasing evidence indicates that FA are essentially utilized as an energy source and are signaling molecules that exert physiological activity of gut microbiota and immune responses. In addition, FA could serve as natural ligands for orphan G protein-coupled receptors (GPCR), also called free fatty acid receptors (FFAR), which intertwine metabolic and immune systems via multiple mechanisms. The present review explores the recent findings on FA absorption and its impact on gut health, particularly addressing the mechanism by which dietary FA potentially influences intestinal microbiota and epithelial functions. Also, this work attempts to uncover research ideas for devising future strategies for manipulating the composition of dietary FA to regulate gut health and support a normal immune system for metabolic and immune disorders.     

High feeding intensity increases the severity of fatty liver in the American mink (Neovison vison) with potential ameliorating role for long-chain n-3 polyunsaturated fatty acids

Background

Rapid body fat mobilization, obesity, and an inadequate supply of n-3 polyunsaturated fatty acids (PUFA) have been suggested to play roles in the etiology of fatty liver in the American mink (Neovison vison). This study examined the effects of feeding intensity and dietary fat source on fatty liver induced by fasting. In a multi-factorial design, 3 different fat sources (herring oil, rich in n-3 PUFA, soya oil, rich in n-6 PUFA, and canola oil, rich in n-9 monounsaturated fatty acids) were fed to mink at a low and high feeding intensity for 10 weeks, followed by an overnight or a 5-day fasting treatment to induce fatty liver.

Results

Fasting led to the development of fatty liver with increased severity in the mink fed at the high feeding intensity. The herring oil diet, high in long-chain n-3 PUFA, was found to decrease the severity of fatty liver in the mink at the high feeding intensity.

Conclusion

Preventing excessive weight gain and increasing dietary intake of n-3 long-chain PUFA may help prevent excessive lipid accumulation during prolonged periods of fasting or inappetence by promoting hepatic fatty acid oxidation.     

Effect of short‐chain fatty acids on triacylglycerol accumulation,lipid droplet formation and lipogenic gene expression in goat mammary epithelial cells

Short‐chain fatty acids (SCFAs) are the major energy sources for ruminants and are known to regulate various physiological functions in other species. However, their roles in ruminant milk fat metabolism are still unclear. In this study, goat mammary gland epithelial cells (GMECs) were treated with 3 mmol/L acetate, propionate or butyrate for 24 h to assess their effects on lipogenesis. Data revealed that the content of triacylglycerol (TAG) and lipid droplet formation were significantly stimulated by propionate and butyrate. The expression of FABP3, SCD1, PPARG, SREBP1, DGAT1, AGPAT6 and ADRP were upregulated by propionate and butyrate treatment. In contrast, the messenger RNA (mRNA) expression of FASN and LXRα was not affected by propionate, but reduced by butyrate. Acetate had no obvious effect on the content of TAG and lipid droplets but increased the mRNA expression of SCD1 and FABP3 in GMECs. Additionally, it was observed that propionate significantly increased the relative content of mono‐unsaturated fatty acids (C18:1 and C16:1) at the expense of decreased saturated fatty acids (C16:0 and C18:0). Butyrate and acetate had no significant effect on fatty acid composition. Overall, the results from this work help enhance our understanding of the regulatory role of SCFAs on goat mammary cell lipid metabolism.     

短链脂肪酸在生猪产业的研究与应用

短链脂肪酸是肠道微生物发酵产物,其对机体健康的影响已被广泛研究。本文就短链脂肪酸在国内外的研究进展,综述了其对猪肠道健康和代谢健康的有益影响以及在生猪养殖中的应用,旨在为新型饲料添加剂的开发与利用提供应用和理论依据。     

Sexual maturation in hens is not associated with increases in serum leptin and the expression of leptin receptor mRNA in hypothalamus

Background

In mammals, leptin is an attractive candidate for mediating the metabolic signal and the reproductive function via the specific receptor in hypothalamus. However, till now, the role of leptin on reproduction in birds is less well established. This experiment was conducted to elucidate the role of leptin on the onset of reproduction in bird, as a first step, to detect the changes of peripheral leptin and leptin receptor mRNA expression in hypothalamus between mature and immature hens at the same age. 120 ISA brown pullets at D60 were allocated randomly into two groups, long light (LL) group being raised under artificial light regimes with incrementally increased light phase (from 8 L:16D to 14 L:12D) and short light (SL) group raised on consistent light (8 L:16D) for 12 wk.

Results

The results showed that pullets in LL group reached sexual maturation 15 d earlier than those in SL group. Serum E₂ showed a significant increase with age, but no difference was observed between two groups. Serum leptin concentration decreased significantly from D112 to D136 in LL, and was markedly higher in LL group than that in SL at D112, while there was no significant difference between two groups at D136. Leptin receptor and GnRH-I mRNA expression in hypothalamus were significantly increased with age, yet there was no significant difference between SL and LL chickens at the same age. The expression of FSH-β and LH-β mRNA in pituitary was increased with age but did not show significant difference between LL and SL group. GnRH-I mRNA expression was very rich in pineal gland, and decreased from D112 to D136 in LL but not in SL group, and there was no difference between two groups at the same age.

Conclusions

These results indicate that the earlier sexual maturation in hens induced by long-light regime is not accompanied with an increase in serum leptin or leptin receptor gene expression in hypothalamus, or genes expression in HPG axis.     

中链脂肪酸在仔猪生产中的研究与应用

中链脂肪酸是一类含有6~12个碳原子的饱和脂肪酸,广泛存在于天然食物中,其对机体具有广泛的有益作用。本文就中链脂肪酸的消化吸收代谢特性、抗菌特性、抗炎抗氧化作用以及在仔猪生产中的应用进行综述,旨在为新型饲料添加剂的开发与利用提供应用和理论依据。     

Dynamic changes of fatty acids and minerals in sow milk during lactation

The objective of the present study was to explore the changes in fatty acids (FAs) and minerals composition in sow milk in order to improve the knowledge about FAs and mineral requirements for piglets. The FAs and minerals composition in the sow milk samples which were collected from ten sows during a period of 16 days of lactation was analysed. The proportion of FAs in sow milk has a significant increase in C8:0, C10:0, C12:0, C14:0, C16:0, C14:1 and C16:1 FAs and the sum of saturated fatty acids (SFAs) according to the lactation period (p < 0.05). The proportion of C18:2 cis‐9,12 FA and the sum of polyunsaturated fatty acids (PUFAs) significantly decreased with the advancing of lactation (p < 0.05). The concentrations of minerals, including potassium, copper, manganese and zinc, were higher on day 1 and day 2 than those on day 13 and day 16 (p < 0.05). On the contrary, the lowest concentrations of calcium, sodium, magnesium and iron and the lowest molar calcium: phosphorus ratio emerged in colostrum (p < 0.05). In summary, our results demonstrated FAs profile and minerals concentrations were changed with lactation period. Better understanding of the changes of FAs and minerals may be valuable to swine nutritionists in the commercial industry. In addition, those results provided some meaningful information for sow's diet formulation during lactation.     

Sexual maturation in hens is not associated with increases in serum leptin and the expression of leptin receptor mRNA in hjpothalamus

Background： In mammals, leptin is an attractive candidate for mediating the metabolic signal and the reproductive function via the specific receptor in hypothalamus. However, till now, the role of leptin on reproduction in birds is less well established. This experiment was conducted to elucidate the role of leptin on the onset of reproduction in bird, as a first step, to detect the changes of peripheral leptin and leptin receptor mRNA expression in hypothalamus between mature and immature hens at the same age. 120 ISA brown pullets at D60 were allocated randomly into two groups, long light （LL） group being raised under artificial light regimes with incrementally increased light phase （from 8 L：]6D to 14 L：]2D） and short light （SL） group raised on consistent light （8 L：16D） for 12 wk. Results： The results showed that pullets in LL group reached sexual maturation 15 d earlier than those in SL group. Serum E2 showed a significant increase with age, but no difference was observed between two groups. Serum leptin concentration decreased significantly from D112 to D136 in LL, and was markedly higher in LL group than that in SL at D112, while there was no significant difference between two groups at D136. Leptin receptor and GnRH-I mRNA expression in hypothalamus were significantly increased with age, yet there was no significant difference between SL and LL chickens at the same age. The expression of FSH-13 and LH-13 mRNA in pituitary was increased with age but did not show significant difference between LL and gland, and decreased from D112 to D136 in LL but not groups at the same age. SL group. GnfiH-I mRNA expression was very rich in pinea n SL group, and there was no difference between two Conclusions： These results indicate that the earlier sexual maturation in hens induced by long-light regime is not accompanied with an increase in serum leptin or leptin receptor gene expression in hypothalamus, or genes expression in HPG axis.     

日粮中添加牛至精油对牛肉脂肪酸以及血液生化指标的影响

为研究日粮中添加牛至精油对牛肉脂肪酸以及血液生化指标含量的影响,试验选择10头体重相近、健康的荷斯坦奶公牛,随机平均分成试验组(每头添加牛至精油28 g/d)和对照组(未添加牛至精油),每组5头,相同环境下饲喂8个月后屠宰。结果显示:添加牛至精油组血液中总蛋白、白蛋白显著升高(P < 0.05);甘油三酯显著降低(P < 0.05);对半腱肌饱和脂肪酸(SFA)和不饱和脂肪酸(UFA)均无显著影响(P > 0.05),但使多不饱和脂肪酸(PUFA)含量显著升高(P < 0.05),P/S值增大(P < 0.05),显著提高了油酸以及亚油酸含量。因此日粮中添加牛至精油可有效改善牛肉脂肪酸组成含量和血液生化指标。     

Interactions between dietary fatty acids and hepatic gene expression in livers of pigs during the weaning period

The aim of the present study was to evaluate the impact of dietary fatty acids (FA) during the weaning period on expression of genes involved in the oxidation and metabolism of FA (peroxisome proliferator-activated receptor (PPAR-), stearoyl-CoA-desaturase (SCD), Δ6-desaturase (D6D), acetyl CoA carboxylase (ACC) and fatty acid synthase (FAS)). Liver samples were obtained from littermates, either on day 28 of age just before weaning, or at day 56 after the 4 weeks ad lib. provision of 5% of either animal fat, fish oil or sunflower oil. In conclusion, genes involved in the regulation of FA conversion (SCD, D6D) were influenced by the n-6 to n-3 ratio, whereas the FA oxidation, as indicated by the expression of PPAR-, was highly likely regulated by the hepatic ratio between mono- and poly-unsaturated FA. Furthermore, weaning and/or age affected the hepatic expression of genes involved in FA synthesis and conversion, but not the expression of PPAR-.     

脂联素对犊牛肝细胞脂氧化关键酶表达的影响

体外原代培养犊牛肝细胞,添加0、16、64和128μg/L脂联素（adiponectin,ADPN）,分别培养4、8h后,提取细胞总RNA。应用实时荧光定量PCR方法,检测脂氧化关键酶脂酰辅酶A氧化酶（ACO）、肝脏脂肪酸结合蛋白（L-FABP）、肉碱脂酰转移酶-Ⅰ（CPTⅠ）、肉碱脂酰转移酶-Ⅱ（CPTⅡ）mRNA表达变化。结果显示,在ADPN作用4h后,ACO、L-FABP、CPTⅠ、CPTⅡ基因mRNA水平均随ADPN浓度的增加而增加,且均显著高于对照组,呈剂量依赖关系;作用8h时,ACO表达也增加,但差异不显著,L-FABP在中高剂量组显著高于对照组,CPTⅠ和CPTⅡ在各ADPN处理组均显著高于对照组。结果表明,ADPN可显著促进脂氧化关键酶的表达,增强肝脏脂氧化作用,减少肝脂储存。     

美仁牦牛肉不同部位常规营养成分及氨基酸和脂肪酸测定分析

摘 要：选取放牧条件下体况良好、体重相近的甘南成年美仁公牦牛 6 头、母牦牛 4头进行屠宰,采取美仁牦牛肉的肩甲、背最长肌、股二头肌三个部位的分割肉。测定不同部位营养成分、氨基酸和脂肪酸。 结果显示：不同部位美仁牦牛肉的蛋白质含量为22.97%～23.07%,其中股二头肌蛋白质含量最低,背最长肌最高;脂肪含量为1.27%～2.49%,股二头肌脂肪脂肪含量较低,不足1.27%。氨基酸含量丰富,脂肪酸总量依次为肩甲、背最长肌、股二头肌上脑肉。通过对比分析不同部位美仁牦牛肉中营养成分、氨基酸和脂肪酸等含量存在差异。