The Pathological Factors Involved in Current In Vitro Atherosclerotic Models.

Cardiovascular disease stemmed from atherosclerosis (AS) is well recognized to be the predominant cause of global death. To comprehensively clarify the pathogenesis of AS, exploit effective drugs, as well as develop therapeutic solutions, various atherosclerotic models were constructed in vitro and widely utilized by the scientific community. Compared with animal models, the in vitro atherosclerotic models play a prominent role not only in the targeted research of single pathological factor related to AS in the human derived system, but also in the combined study on multipathological factors leading to AS, thereby contributing tremendously to the in-depth elucidation of atherosclerotic pathological process. In the current review, a variety of pathological factors incorporated into the existing atherosclerotic models in vitro are broadly elaborated, including the pathological mechanism, in vitro simulation approaches, and the desired improvement perspectives for reproducing each pathological factor. In addition, this review also summarizes the advantages and disadvantages of current atherosclerotic models as well as their potential functionality. Finally, the promising aspects for future atherosclerotic models in vitro with potential advances are also discussed.

Blue LED light induces cytotoxicity via ROS production and mitochondrial damage in bovine subcutaneous preadipocytes.

The purpose of this study was to investigate the effect and mechanism of blue light irradiation on bovine subcutaneous preadipocytes. In this study, preadipocytes were divided into dark group (control) and blue light group. Results show that blue light exposure time-dependently reduced the viability of preadipocytes and induced mitochondrial damage, in accompaniment with the accumulation of intracellular reactive oxygen species (ROS). Meanwhile, blue light caused oxidative stress, as evidenced by the increased MDA level, the reduced T-AOC contents, as well as the decreased activities of antioxidant enzymes. Additionally, blue light treatment induced apoptosis and G2/M phase arrest via Bcl-2/Bax/cleaved caspase-3 pathway and P53/GADD45 pathway, respectively. Protein expressions of LC3-II/LC3-I and P62 were up-regulated under blue light exposure, indicating blue light initiated autophagy but impeded autophagic degradation. Moreover, blue light caused an increase in the secretion of pro-inflammatory factors (TNF-α, IL-1ß, and IL-6). Pretreatment with N-acetylcysteine (NAC), a potent ROS scavenger, restored the loss of mitochondrial membrane potential (Δψ) and reduced excess ROS. Additionally, the above negative effects of blue light on cells were alleviated after NAC administration. In conclusion, this study demonstrates blue light induces cellular ROS overproduction and Δψ depolarization, resulting in the decrease of cell viability and the activation of apoptosis, autophagy, and inflammation, providing a reference for the application of blue light in the regulation of fat cells in the future.

Cold exposure induces browning of bovine subcutaneous white fat in vivo and in vitro.

White adipocytes can be transformed into beige adipocytes through the process of browning under cold exposure. To investigate the effects and underlying mechanisms of cold exposure on subcutaneous white fat in cattle, in vitro and in vivo studies were performed. Eight bulls of Jinjiang cattle breed (Bos taurus) aged 18 months were allocated to the control group (n = 4, autumn) or the cold group (n = 4, winter) by different slaughter seasons. Biochemical and histomorphological parameters were detected in blood and backfat samples. Subcutaneous adipocytes from Simental cattle (Bos taurus) were then isolated and cultured at a normal body temperature (37 °C) and at a cold temperature (31 °C) in vitro. In the in vivo study, cold exposure stimulated subcutaneous white adipose tissue (sWAT) browning by reducing adipocyte sizes and up-regulating the expression levels of browning-specific makers (UCP1, PRDM16, and PGC-1α) in cattle. In addition, cold-exposed cattle displayed lower lipogenesis transcriptional regulator levels (PPARγ and CEBPα) and higher lipolysis regulator levels (HSL) in sWAT. In the in vitro study, cold temperature inhibited subcutaneous white adipocytes (sWA) adipogenic differentiation by reducing lipid contents and decreasing the expression of adipogenic marker genes and proteins. Furthermore, cold temperature led to sWA browning which was characterized by increased browning-related genes, mitochondrial contents, and mitochondrial biogenesis-specific markers. In addition, p38 MAPK signaling pathway activity was stimulated by the incubation in cold temperature for 6 h in sWA. We concluded that the cold-induced browning of the subcutaneous white fat was beneficial to the production of heat and the maintenance of body temperature regulation in cattle.

Low temperature exposure inhibits proliferation and induces apoptosis of bovine subcutaneous preadipocytes via p38 MAPK/JNK activation.

Cold stress can cause changes in cell growth states. Cell proliferation and apoptosis are important physiological processes to maintain normal growth and development of cells and tissues. We sought to identify the impact of low temperature exposure on bovine subcutaneous preadipocyte proliferation and apoptosis. We first cultured preadipocytes at different low temperatures (35°C, 33°C, and 31°C) for 12 h, and then at 31°C for 24 and 48 h. The results showed that compared with the 37°C group, exposure to 31°C significantly reduced cell viability and number as well as inhibited cell cycle progression in preadipocytes. Moreover, low temperature also significantly upregulated the apoptosis rate of preadipocytes. After low temperature treatment, the mRNA levels of Cyclin E, CDK2 and B-cell lymphoma 2 (Bcl-2) were decreased in preadipocytes, whereas that of p53, p21 and Bcl-2 associated x protein (Bax) were increased in preadipocytes. Concurrently, low temperature increased the proteins levels of p53, Bax and Cleaved Caspase3, and reduced the protein level of Bcl-2 in preadipocytes. Furthermore, the elevated phosphorylation levels of p38 mitogen-activated protein kinases (p38 MAPK) and c-jun NH2-terminal kinase (JNK) were detected in cold-treated preadipocytes. The influence of low temperature exposure on preadipocyte proliferation and apoptosis were obviously weakened after blocking the p38 MAPK and JNK signaling pathways. In conclusion, low temperature exposure could inhibit proliferation and cell cycle progression and induce apoptosis through activation of p38 MAPK and JNK signaling pathways in bovine subcutaneous preadipocytes.

Omics analysis of the effect of cold normal saline stress through gastric gavage on LPS induced mice.

Cold stress is a significant environmental stimulus that negatively affects the health, production, and welfare of animals and birds. However, the specific effects of cold stimulation combined with lipopolysaccharide (LPS) on the mouse intestine remain poorly understood. Therefore, we designed this research to explore the effect of cold stimulation + LPS on mice intestine via microbiome and microbiota sequencing. Forty-eight mice were randomly divided into four experimental groups (n = 12): Control (CC), LPS-induced (CL), cold normal saline-induced (MC) and LPS + cold normal saline-induced (ML). Our results showed body weight was similar among different groups of mice. However, the body weight of mice in groups CC and CL were slightly higher compared to those in groups MC and ML. The results of gene expressions reflected that CL and ML exposure caused gut injury and barrier dysfunction, as evident by decreased ZO-1, OCCLUDIN (P < 0.01), and CASPASE-1 (P < 0.01) expression in the intestine of mice. Moreover, we found that cold stress induced oxidative stress in LPS-challenged mice by increasing malondialdehyde (MDA) accumulation and decreasing the antioxidant capacity [glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), total and antioxidant capacity (T-AOC)]. The cold stress promoted inflammatory response by increased IL-1ß in mice treated with cold normal saline + LPS. Whereas, microbiome sequencing revealed differential abundance in four phyla and 24 genera among the mouse groups. Metabolism analysis demonstrated the presence of 4,320 metabolites in mice, with 43 up-regulated and 19 down-regulated in CC vs. MC animals, as well as 1,046 up-regulated and 428 down-regulated in ML vs. CL animals. It is Concluded that cold stress enhances intestinal damage by disrupting the balance of gut microbiota and metabolites, while our findings contribute in improving management practices of livestock in during cold seasons.

Growth hormone inhibits adipogenic differentiation and induces browning in bovine subcutaneous adipocytes.

OBJECTIVE: It is well established that growth hormone (GH) has the ability to stimulate lipolysis. The effects of GH on adipocyte differentiation and browning have not been clearly described. Therefore, the present study aimed to elucidate the role of GH in the differentiation and browning of bovine subcutaneous adipocytes as well as its underlying molecular mechanisms. METHODS: We first treated bovine subcutaneous preadipocytes with different concentrations (0, 10, 100, and 500 ng/mL) of GH for 8 days and measured lipid accumulation and gene expression. Afterward, we treated preadipocytes and mature adipocytes with 500 ng/mL GH and determined differentiation and browning-related indicators. Finally, we investigated the expression of STAT5B in both preadipocytes and mature adipocytes after GH treatment. RESULTS: We demonstrated that GH inhibited lipid accumulation and decreased the expression levels of adipogenic key genes (SCD1, SREBP1, PPARγ, and CEBPα) during adipocyte differentiation. Moreover, we observed that the inhibitory effect of GH on the early stage of adipocyte differentiation (0-2 days) was stronger than that on the later stage of adipocyte differentiation (2-8 days). We also found that GH promoted the expression levels of browning-related genes such as uncoupling protein 1 (UCP1) in mature adipocytes. Concurrently, GH promoted mitochondrial biogenesis and increased the expression levels of mitochondrial biogenesis-related genes. In addition, GH promoted phosphorylation of signal transducers and activator of transcription 5 b (STAT5B) and contributed to translocation of STAT5B to nucleus. After blocking the expression of STAT5B protein, GH weakened the inhibition of adipogenic key genes and reduced the promotion of browning-related genes in bovine subcutaneous adipocytes. CONCLUSIONS: GH can inhibit adipocyte differentiation and promote adipocyte browning by regulating STAT5B in bovine subcutaneous adipocytes.

Procyanidin B2 Alleviates Heat-Induced Oxidative Stress through the Nrf2 Pathway in Bovine Mammary Epithelial Cells.

The objective of this study was to investigate the protective effects and potential molecular mechanisms of procyanidin B2 (PB2) in MAC-T (mammary alveolar cells-large T antigen) cells during heat stress (HS). The MAC-T cells were divided into three treatment groups: control (37 °C), HS (42 °C), and PB2 + HS (42 °C). Compared with MAC-T cells that were consistently cultured at 37 °C, acute HS treatment remarkably decreased cell viability, reduced activities of catalase (CAT), superoxide dismutase (SOD), and total antioxidant capacity (T-AOC), and elevated intracellular levels of malondialdehyde (MDA) and reactive oxygen species (ROS). Additionally, nuclear factor erythroid 2-related factor 2 (Nrf2) was activated and translocated to the nucleus, in accompaniment with upregulation of Nrf2, heme oxygenase 1 (HO-1), thioredoxin reductase 1 (Txnrd1), and heat shock protein 70 (HSP70). In parallel, both mRNA transcript and actual protein secretion of pro-inflammatory cytokines, including tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß), were increased by heat stress. Pretreatment of MAC-T cells with 0~25 µM PB2 alleviated the decline of cell viability by HS in a dose-dependent fashion and protected cells against HS-induced oxidative stress, as evidenced by significantly improved CAT, SOD, and T-AOC activity, as well as with decreased MDA and ROS generation. Furthermore, PB2 further activated the Nrf2 signaling pathway and reversed the inflammatory response induced by HS. Silencing of Nrf2 by si-Nrf2 transfection not only exacerbated HS-induced cell death and provoked oxidative stress and the inflammatory response, but also greatly abolished the cytoprotective effects under HS of PB2. In summary, PB2 protected MAC-T cells against HS-induced cell death, oxidative stress, and inflammatory response, partially by operating at the Nrf2 signal pathway.

Association between high serum Nogo-B and hypertension in Chinese Han.

BACKGROUND: Cellular and animal studies have shown that endoplasmic reticulum protein B (Nogo-B) is associated with hypertension, but that association has not been fully studied in humans. Therefore, the expression levels of Nogo-B were investigated in hypertensive patients. METHODS: The plasma Nogo-B levels of 74 patients with hypertension and 67 non-hypertensive patients were measured by enzyme-linked immunosorbent assay. RESULTS: The plasma Nogo-B levels in the hypertensive group [523.43(411.41-746.79)] were higher than in the non-hypertensive group [380.29(281.57-462.13)] (P < 0.01). Pearson's correlation analysis indicated that systolic blood pressure and diastolic blood pressure were linearly and positively correlated with plasma Nogo-B levels. Multivariable logistic regression analysis was performed based on sex, age, BMI, smoking history, drinking history, and levels of TC, TG, LDL, and HDL. The results indicated that the plasma Nogo-B levels were independently associated with hypertension (OR = 1.007, 95%CI: 1.004-1.010, P < 0.01). CONCLUSIONS: The present study suggests that hypertensive participants exhibited higher plasma Nogo-B levels than those without hypertension. Plasma Nogo-B levels are independently associated with hypertension.

Effects of Backfat Thickness on Oxidative Stress and Inflammation of Placenta in Large White Pigs.

The purpose of this study was to evaluate the impact of the backfat thickness of sows on reproductive performance and on lipid metabolism, oxidative stress, and inflammation. At farrowing, 60 sows were assigned to three groups: the low-backfat-thickness group (LBF, n = 20): sows' backfat thickness was between 9 and 12 mm; the medium-backfat-thickness group (MBF, n = 20): sows' backfat thickness was between 13 and 20 mm; and the high-backfat-thickness group (HBF, n = 20): sows' backfat thickness was between 21 and 25 mm. Maternal and fetal blood and placental samples were collected. Compared with the LBF and HBF groups, the MBF group delivered a significantly greater number of live piglets than the LBF or HBF groups. The different backfat thicknesses of sows had different effects on the lipid-related hormones and adipokines of maternal and fetal serum and placenta. Sows with poor or excessive backfat displayed higher levels of oxidative stress and higher levels of pro-inflammatory cytokines. According to these data, the thickness of a sow's backfat affects the characteristics of farrowing piglets and their lipid metabolism, as well as placental inflammation, maternal inflammation, and oxidative stress. A moderate backfat thickness (between 13 and 20 mm) was associated with greater reproductive performance in sows.

Preconditioning with procyanidin B2 protects MAC-T cells against heat exposure-induced mitochondrial dysfunction and inflammation.

Heat stress (HS) induced by high environmental temperature is a main factor causing mastitis and reduced milk production in dairy cows. Procyanidin B2 (PB2) is a phenolic compound with strong anti-inflammatory and antioxidant properties. By using the MAC-T (mammary alveolar cells-large T antigen) cells as the in vitro cell model, this study determines PB2 effects on HS-induced MAC-T mitochondrial dysfunction, cell apoptosis, and inflammation. Cells were divided into three groups: Con (37 °C), HS (42 °C), and PB2 +HS. Results show that, under HS-exposure, MAC-T cells exhibited an increased accumulation of reactive oxygen species (ROS) and Ca2+, a decreased mitochondrial membrane potential (Δψ) and ATP content. Besides, HS markedly induced cell apoptosis, as evidenced by flow cytometry and significantly increased mRNA and protein expressions of apoptosis-related genes, including cytochrome C (Cyto-c) and cleaved caspase-3, etc. HS also led to mitochondrial fission and fusion dynamic disorder. Meanwhile, HS induced a significant inflammatory response by activating the Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κß) signaling pathway and the NOD-like receptor with pyrin domain containing-3 (NLRP3) inflammasome. Notably, preconditioning of PB2 alleviated the accumulation of ROS and Ca2+ concentration induced by HS, increased Δψ and ATP content, and maintained the dynamic balance of mitochondrial fission and fusion, thus improving mitochondrial function. PB2 also blocked the HS-induced mitochondrial caspase apoptosis pathway. Furthermore, PB2 preconditioning inhibited HS-induced activation of the TLR4/NF-κß signaling pathway and the NLRP3 inflammasome, as well as IL-1ß release, reversing HS-induced inflammation. In conclusion, PB2 has an important protective effect against the mitochondrial dysfunction, inflammatory response, and apoptosis of MAC-T cells induced by HS.

BMP2 increases hyperplasia and hypertrophy of bovine subcutaneous preadipocytes via BMP/SMAD signaling.

The study aims to characterize functions of bone morphogenetic protein 2 (BMP2) gene in the process of subcutaneous (SQ) fat deposition of bovine, thereby providing insights into mechanisms for the use of BMP2 in fat management. Our results show that BMP2 was extensively expressed in bovine and relatively rich in adipose tissue. Exogenous BMP2 significantly enhanced proliferation of bovine preadipocytes. Consistently, si-BMP2 apparently induced cell cycle arrest at G0/G1 phase and decreased proliferation of preadipocytes. Meanwhile, exogenous BMP2 mildly enhanced preadipocyte differentiation at day 3 of differentiation, as evidenced by accelerated lipid accumulation, as well as increased mRNA and protein expressions of adipogenic key transcription factor PPARγ; contrary results about lipids were found by BMP2 interference treatment. No difference was observed concerning BMP2 or si-BMP2 treatment at day - 2 and day 0 of differentiation. Additionally, LDN-193189 (inhibitor of BMP type I receptor) pretreatment diminished the enhancement of preadipocyte proliferation and differentiation induced by BMP2, as evidenced by constant proliferation rate and PPARγ expressions. Furthermore, BMP2 markedly enhanced phosphorylation level of SMAD1/5/9, and LDN-193189 could diminish the difference caused by BMP2. Thus, our results suggest that BMP2 triggers BMP/SMAD signaling pathway, promoting both hyperplasia and hypertrophy of bovine preadipocytes.

Microtubule Affinity-Regulating Kinase 4 Promotes Oxidative Stress and Mitochondrial Dysfunction by Activating NF-κB and Inhibiting AMPK Pathways in Porcine Placental Trophoblasts.

The aim of this investigation was to evaluate the role of MARK4 in the regulation of oxidative stress and mitochondrial dysfunction in pig placental trophoblasts and analyze the signaling pathways involved. In this study, we found that enhanced MARK4 contributed to augmented oxidative stress in pig trophoblasts, as evidenced by decreased total antioxidant capacity (TAC); higher production of reactive oxygen species (ROS); elevated protein carbonylation; and reduced SOD, CAT, and GSH-PX activities. Further analyses revealed MARK4 impaired mitochondrial oxidative respiration in cultured trophoblasts, which was associated with reduced ATP content, decreased mitochondrial membrane potential, lower mitochondrial Complexes I and III activities, and down-regulated protein contents of subunits of complexes I, II, and V. At same time, mitochondrial biogenesis and structure were negatively altered by elevated MARK4. By antioxidant treatment with vitamin E (VE), oxidative stress along with impaired mitochondrial function induced by enhanced MARK4 were blocked. Furthermore, we found activation of AMPK signaling prevented MARK4 from blocking mitochondrial biogenesis and function in pig trophoblast cells. Finally, we demonstrated that the IKKα/NF-κB signal pathway was involved in MARK4 activated oxidative stress and mitochondrial dysfunction. Thus, these data suggest that MARK4 promotes oxidative stress and mitochondrial injury in porcine placental trophoblasts and can contribute to the developing of knowledge of pathological processes leading to mitochondrial dysfunction associated with excessive back-fat in the pig placenta and to the obesity-associated pregnant syndrome.

Effects of prolonged photoperiod on growth performance, serum lipids and meat quality of Jinjiang cattle in winter.

OBJECTIVE: This study was conducted to investigate the potential effects of prolonged photoperiod on the serum lipids, carcass traits, and meat quality of Jinjiang cattle during winter. METHODS: Thirty-four Jinjiang bulls aged between 14 and 16 months were randomly assigned to two groups that were alternatively subjected to either natural daylight +4 h supplemental light (long photoperiod, LP) or natural daylight (natural photoperiod, NP) for 96 days. The potential effects on the levels of serum lipids, carcass traits, meat quality, and genes regulating lipid metabolism in the intramuscular fat (IMF) of the cattle were evaluated. RESULTS: Jinjiang cattle kept under LP showed significant increase in both dry matter intake and backfat thickness. the serum glucose and the plasma leptin levels were significantly reduced, while that of melatonin and insulin were observed to be increased. The crude fat contents of biceps femoris muscle and longissimus dorsi muscle were higher in LP than in NP group. In longissimus dorsi muscle, the proportions of C17:0 and C18:0 were significantly higher but that of the C16:1 was found to be significantly lower in LP group. The relative mRNA expressions in IMF of longissimus dorsi muscle, the lipid synthesis genes (proliferatoractivated receptor gamma, fatty acid-binding protein) and the fatty acid synthesis genes (acetyl-coa carboxylase, fatty acid synthetase, 1-acylglycerol-3-phosphate acyltransferase) were significantly up-regulated in LP group (p<0.05); whereas the hormone-sensitive lipase and stearoyl-CoA desaturase 1 were significantly down-regulated in LP than in NP group. CONCLUSION: Prolonged photoperiod significantly altered the growth performance, hormonal levels, gene expression and fat deposition in Jinjiang cattle. It suggested that the LP improved the fat deposition by regulating the levels of different hormones and genes related to lipid metabolism, thereby improving the fattening of Jinjiang cattle during winter.

Cytoprotective Effects of Taurine on Heat-Induced Bovine Mammary Epithelial Cells In Vitro.

It is a widely known that heat stress induces a reduction in milk production in cows and impairs their overall health. Studies have shown that taurine protects tissues and organs under heat stress. However, there have yet to be studies showing the functions of taurine in mammary alveolar cells-large T antigen (MAC-T) (a bovine mammary epithelial cell line) cells under heat shock. Therefore, different concentrations of taurine (10 mM, 50 mM, and 100 mM) were tested to determine the effects on heat-induced MAC-T cells. The results showed that taurine protected the cells against heat-induced damage as shown by morphological observations in conjunction with suppressed the translocation and expression of heat shock factor 1 (HSF1). Moreover, taurine not only reversed the decline in antioxidase (superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX)) activities but also attenuated the accumulation of malondialdehyde (MDA). Meanwhile, mitochondrial damage (morphology and complex I activity) resulting from heat exposure was mitigated. Taurine also alleviated the rates of cell apoptosis and markedly depressed the mRNA expressions of BCL2 associated X, apoptosis regulator (BAX) and caspase3. Furthermore, compared with the heat stress (HS) group, the protein levels of caspase3 and cleaved caspase3 were decreased in all taurine groups. In summary, taurine improves the antioxidant and anti-apoptosis ability of MAC-T cells thereby alleviates damage of cells due to heat insults.

Validity of five formulas in estimating 24-h urinary sodium via spot urine sampling in hypertensive patients living in Northeast China.

OBJECTIVE: The objective was to evaluate the accuracy of five formulas -- the Kawasaki, Tanaka, INTERSALT, Mage, and Uechi methods -- using spot urinary sampling for 24-h urinary sodium (UNa) prediction in hypertensive patients living in northeast China. METHODS: There were 1154 hypertensive patients enrolled from multiple centers. Five different formulas were used to predict 24-h UNa excretion via spot morning urinary samples. Actual UNa excretion was measured from 24-h urine samples. The estimated value was compared with the actual value by examining biases, the intraclass correlation coefficients (ICC), and Bland-Altman plots. RESULTS: The average excretion of sodium was 2.97 ±â€Š1.26 g/day. The formula-produced mean biases for actual UNa were 0.31 g/day for INTERSALT, 0.80 g/day for Mage, 0.88 g/day for Tanaka, 1.14 g/day for Uechi, and 1.95 g/day for Kawasaki. The ICC was 0.511 for Kawasaki, 0.499 for INTERSALT, 0.468 for Tanaka, 0.402 for Mage, and 0.378 for Uechi. The least mean bias in the lower and moderate salt intake subgroups was 1.22 and 0.07 g/day, respectively, which was calculated using the Mage and INTERSALT methods. The least mean bias in the higher salt intake subgroup was 0.10 g/day for the Uechi method. The INTERSALT method was more efficiency at the individual level, with 17.4% of participants having relative differences within 10%, and 22.3% participants having absolute differences within 393 mg. CONCLUSION: The INTERSALT method may exhibit a good performance in estimating 24-h urinary sodium level for the hypertensive population living in northeast China.

Impaired Mitochondrial Function Results from Oxidative Stress in the Full-Term Placenta of Sows with Excessive Back-Fat.

The aim of this study was to determine the effect of excessive back-fat (BF) of sows on placental oxidative stress, ATP generation, mitochondrial alterations in content and structure, and mitochondrial function in isolated trophoblasts. Placental tissue was collected by vaginal delivery from BFI (15-20 mm, n = 10) and BFII (21-27 mm, n = 10) sows formed according to BF at mating. Our results demonstrated that excessive back-fat contributed to augmented oxidative stress in term placenta, as evidenced by excessive production of ROS, elevated protein carbonylation, and reduced SOD, GSH-PX, and CAT activities (p < 0.05). Indicative of mitochondrial dysfunction, reduced mitochondrial respiration in cultured trophoblasts was linked to decreased ATP generation, lower mitochondrial Complex I activity and reduced expression of electron transport chain subunits in placenta of BFII sows (p < 0.05). Meanwhile, we observed negative alterations in mitochondrial biogenesis and structure in the placenta from BFII group (p < 0.05). Finally, our in vitro studies showed lipid-induced ROS production resulted in mitochondrial alterations in trophoblasts, and these effects were blocked by antioxidant treatment. Together, these data reveal that excessive back-fat aggravates mitochondrial injury induced by increased oxidative stress in pig term placenta, which may have detrimental consequences on placental function and therefore impaired fetal growth and development.

The Effects of Maternal Obesity on Porcine Placental Efficiency and Proteome.

Maternal obesity is associated with impaired maternal metabolism and affects the developmental programming of the fetus. The placenta is dysfunctional when exposed to an obese intrauterine environment and can transduce and mediate detrimental maternal impacts to the fetus through mechanisms that remain largely unknown. The main objective of this study was to investigate the effects of maternal obesity on the porcine placental proteome and to analyze the deregulated proteins and potential pathways predicted to be disturbed in obese placentas, using sows with high backfat as a model of obese pregnancy. The sows were divided into two groups based on their backfat thickness: normal backfat (NBF, 17-22 mm; n = 30) and high backfat (HBF, ≥23 mm; n = 30) as the maternal obesity group. The placental tissues used for the proteomic and biochemical analyses were obtained through vaginal delivery, and the maternal blood samples used to determine the metabolic parameters were collected at day 107 of pregnancy. Our study demonstrated that HBF sows had significantly decreased placental efficiency, increased plasma-free fatty acids and triglyceride levels, and increased proinflammatory cytokines plasma levels (p < 0.05). HBF placentas had significantly higher malondialdehyde level, lower total antioxidant capacity and antioxidase activity, increased triglyceride content and enhanced proinflammatory tumor necrosis factor- α (TNF-α) and interleukin-6 (IL-6) contents (p < 0.05). Among the 4652 proteins identified using the proteomic method, 343 were quantified as differentially abundant proteins, which were involved in many vital biological processes. Based on our bioinformatic and placental biochemical analyses, we concluded that maternal obesity is associated with abnormal carbohydrate and lipid metabolism, mitochondrial dysfunction, decreased steroid hormone biosynthesis, and increased oxidative stress and inflammation in the placenta. The results of this study are undoubtedly valuable to other researchers.

Molecular Characterization of Microtubule Affinity-Regulating Kinase4 from Sus scrofa and Promotion of Lipogenesis in Primary Porcine Placental Trophoblasts.

This study aimed to characterize the full-length cDNA of MARK4 in Sus scrofa, and evaluated its potential role in the regulation of lipid accumulation in pig placental trophoblasts and analyzed signaling pathways involved, thereby providing insights into mechanisms for placental lipotoxicity induced by excessive back-fat during pregnancy of sows. The cDNA obtained with 5' and 3' RACE amplification covered 3216 bp with an open reading frame of 2259 bp encoding 752 amino acids. Multiple alignments and phylogenetic analysis revealed MARK4 protein of Sus scrofa had a high homology (95%⁻99%) to that of other higher vertebrates. After transfection, enhanced MARK4 significantly promoted lipogenesis in pig trophoblasts, as evidenced by accelerated lipid accumulation and consistently increased mRNA expressions of lipogenic genes DGAT1, LPIN1, LPIN3, LPL, PPARδ and SREBP-1c. Meanwhile, PPARγ remarkably inhibited the stimulating effect of MARK4 on non-receptor-mediated lipid accumulation in trophoblasts. Further analyses revealed WNT signaling enhanced lipid accumulation and activation of MARK4 in pig trophoblast cells. Finally, we demonstrated that WNT/ß-catenin signal pathway is involved in MARK4 activated lipogenesis. These results suggest that MARK4 promotes lipid accumulation in porcine placental trophoblasts and can be considered as a potential regulator of lipotoxicity associated with maternal obesity in the pig placenta.

ZBTB16 Overexpression Enhances White Adipogenesis and Induces Brown-Like Adipocyte Formation of Bovine White Intramuscular Preadipocytes.

BACKGROUND/AIMS: Our study aims to characterize functions of ZBTB16 gene in the process of intramuscular fat (IMF) deposition and metabolism of bovine, thereby providing insights into mechanisms for the use of ZBTB16 in fat management. METHODS: Primary preadipocytes derived from bovine IMF tissue were isolated and used as the in vitro cell model. An adenovirus Ad-ZBTB16 was transfected into bovine preadipocytes to overexpress the ZBTB16 gene. By using real-time quantitative PCR (RT-qPCR), western blotting, Oil Red-O staining, glycerol-3-phosphate dehydrogenase (GPDH) activity assay, and cell counting kit-8 (CCK-8) test, adipogenic and proliferative signals in adipocytes were monitored to investigate effects of ZBTB16 on adipogenesis of bovine preadipocytes. RESULTS: After transfection, mRNA and protein levels of ZBTB16 gene were significantly increased. Enhanced ZBTB16 significantly promoted preadipocyte differentiation, as evidenced by accelerated lipid accumulation, enhanced GPDH activity, consistently increased mRNA expressions of adipogenic key transcription factors PPARγ, C/EBPα, FABP4, and ADIPOQ, and markedly increased protein expressions of PPARγ and FABP4. No difference was observed concerning proliferation of preadipocytes after treatment with Ad-ZBTB16. Furthermore, relative mRNA levels of brown adipocyte selective genes (PRDM16, UCP1, Cidea, Cox8b, and PGC-1α) and beige adipocyte selective genes (CD137, TMEM26, and Tbx1) as well as UCP1 protein expression were significantly increased by Ad-ZBTB16. Meanwhile, Ad-ZBTB16 treatment remarkably induced mitochondrial biogenesis and increased relative mitochondrial DNA (mtDNA) copy number in bovine adipocytes. CONCLUSION: These results suggest that ZBTB16 overexpression can promote white adipogenesis and induce brown-like adipocyte formation for bovine white intramuscular preadipocytes.

Estimation of metabolisable energy and net energy of rice straw and wheat straw for beef cattle by indirect calorimetry.

Two experiments were conducted to estimate the metabolisable energy (ME) and net energy (NE) of rice straw and wheat straw for beef cattle. In each experiment, 16 Wandong bulls (Chinese indigenous yellow cattle) were assigned to 4 dietary treatments in a completely randomised design. Four dietary treatments included one corn silage-concentrate basal diet and three test diets in which the basal diet was partly substituted by rice straw (Exp. 1) or wheat straw (Exp. 2) at 100, 300 and 600 g/kg. Total collection of faeces and urine was conducted for 5 consecutive days after a 2-week adaption period, followed by a 4-d period where gas exchange measurements were measured by an open-circuit respiratory cage. Linear regression equations of rice straw- or wheat straw-associated ME and NE contribution in test diets against rice straw or wheat straw substitution amount were developed to predict the ME and NE values of rice straw and wheat straw. These regression equations resulted in ME and NE values (dry matter basis) of 6.76 and 3.42 MJ/kg for rice straw and 6.43 and 3.28 MJ/kg for wheat straw, respectively. The NE and ME requirement for maintenance of Wandong cattle fed a straw-based diet were 357 and 562 kJ·kg-0.75·d-1, respectively. The regression-derived ME and NE have lower standard errors and coefficients of variation than those estimated by any single substitution ratio. Our study found that the regression method based on multiple point substitution is more reliable than the substitution method for energy evaluation of feedstuffs for beef cattle.