丁酸钠通过AMPK通路调控LPS造成牛乳腺上皮细胞脂代谢紊乱的作用机制

doi:10.11843/j.issn.0366-6964.2022.09.036

Abstract

Abstract: This study aimed to investigate the regulation mechanism of (2, 8, 16 μmol·L^-1) sodium butyrate to the LPS-induced lipid metabolism disorder model of bovine mammary epithelial cell line (MAC-T), explore its regulation mechanism on cell lipid metabolism and repair inflammation damage effect. After stimulating MAC-T cells with 1 000 ng·mL^-1 LPS for 9 h, the lipid droplet area and the cell triglyceride (TG) content were measured respectively. After stimulating MAC-T cells with different concentrations of sodium butyrate for 12 h. MAC-T cells were added with different concentrations of sodium butyrate, and the apoptosis rate was detected by flow cytometry. Subsequently, the experiment was divided into 5 groups:control group, LPS treatment group, 2 μmol·L^-1 sodium butyrate + LPS treatment group, 8 μmol·L^-1 sodium butyrate +LPS treatment group and 16 μmol·L^-1 sodium butyrate+LPS treatment group. Cell TG content, AMPK signaling pathway protein, key genes of lipid metabolism and related inflammatory factors were detected. Results:LPS will cause a significant decrease in the total lipid droplet area of MAC-T cells (P<0.05), and extremely significant decrease in TG content (P<0.01). Different concentrations of sodium butyrate had no effect on the apoptosis rate of MAC-T cells. Compared to the control group, LPS treatment group significantly decreased the content of TG (P<0.01), significantly increased the expression level of P-AMPK (P<0.05), lipid anabolism related genes ACC, SCD-1 and FAS mRNA expression levels were significantly (P<0.05) or extremely significantly decreased (P<0.01), lipid catabolism-related genes CPT-1, CPT-2 and ACO mRNA expression levels of were significantly increased (P<0.05), and contents of inflammatory factors TNF-α and IL-6 were significantly increased (P<0.05). Compared with LPS treatment groups, (2, 8, 16 μmol·L^-1) sodium butyrate +LPS treatment groups increased TG content, decreased P-AMPK expression level, increased lipid anabolism related gene expression level, decreased lipid catabolism related gene expression level, and decreased inflammatory factors TNF-α and IL-6 content. This study shows that sodium butyrate activates lipid anabolism through the AMPK pathway, regulates TG synthesis, and alleviates the inflammatory damage in MAC-T cells.

Key words: lipopolysaccharide, sodium butyrate, bovine mammary epithelial cell line (MAC-T), lipid metabolism disorder

CLC Number:

S856.5

LI Lin, CAO Meng, GONG Binbin, ZHAO Mei, WANG Jie, ZHANG Xiaohui. The Mechanism of Sodium Butyrate through AMPK Pathway to Regulate Lipid Metabolism Disorder Caused by LPS in Bovine Mammary Epithelial Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(9): 3221-3230.

References 31

[1]	KHAN I T, NADEEM M, IMRAN M, et al.Antioxidant properties of milk and dairy products:a comprehensive review of the current knowledge[J]. Lipids Health Dis, 2019, 18:41.
[2]	方莉莉, 杨金芳, 荆海霞, 等. 雌激素受体在泌乳中期和静止期牦牛乳腺组织内的分布与表达[J]. 畜牧兽医学报, 2019, 50(1):211-217.FANG L L, YANG J F, JING H X, et al. Distribution and expression of estrogen receptors in mammary gland of yak during mid-lactating period and non-lactating period[J]. Acta Veterinaria et Zootechnica Sinica, 2019, 50(1):211-217. (in Chinese)
[3]	GALLIER S, TOLENAARS L, PROSSER C. Whole goat milk as a source of fat and milk fat globule membrane in infant formula[J]. Nutrients, 2020, 12(11):3486.
[4]	MALDONADO R F, SÁ-CORREIA I, VALVANO M A. Lipopolysaccharide modification in Gram-negative bacteria during chronic infection[J]. FEMS Microbiol Rev, 2016, 40(4):480-493.
[5]	王林枫, 贾少丹, 杨改青, 等. 脂多糖对奶山羊肝脏代谢组学的影响[J]. 中国农业科学, 2015, 48(18):3701-3710.WANG L F, JIA S D, YANG G Q, et al. Study on the effect of lipopolysaccharide on hepatic metabolism in dairy goat liver[J]. Scientia Agricultura Sinica, 2015, 48(18):3701-3710. (in Chinese)
[6]	ZHOU Z H, XU N B, MATEI N, et al. Sodium butyrate attenuated neuronal apoptosis via GPR41/Gβγ/PI3K/Akt pathway after MCAO in rats[J]. J Cereb Blood Flow Metab, 2021, 41(2):267-281.
[7]	吴泳江. 丁酸钠和β-羟丁酸钠缓解瘤胃细菌细胞壁成分对奶牛炎症和泌乳影响的研究[D]. 重庆:西南大学, 2021.WU Y J. A study on the roles of sodium butyrate and β-hydroxybutyrate in mitigating the effects of rumen bacterial cell-wall components on inflammation and lactation in dairy cows[D]. Chongqing:Southwest University, 2021. (in Chinese)
[8]	李林, 李建嫄, 赵梅, 等. 基于AMPK信号通路研究LPS对奶牛乳腺上皮细胞脂代谢的调控机理[J]. 畜牧与兽医, 2021, 53(12):41-48.LI L, LI J Y, ZHAO M, et al. Determination of the regulation mechanism of LPS on lipid metabolism in bovine mammary epithelial cells based on the AMPK signaling pathway[J]. Animal Husbandry & Veterinary Medicine, 2021, 53(12):41-48. (in Chinese)
[9]	宋美洁, 欧爱群, 薛晓锋, 等. 蜂胶提取物对脂多糖诱导小鼠急性乳腺炎及乳腺屏障功能的保护作用[J]. 中国农业科学, 2021, 54(12):2675-2688.SONG M J, OU A Q, XUE X F, et al. Protective effects of Chinese Propolis extract against lipopolysaccharide- induced acute mastitis and mammary barrier functions in mice[J]. Scientia Agricultura Sinica, 2021, 54(12):2675-2688. (in Chinese)
[10]	RAN X, ZHANG Y, YANG Y X, et al. Dioscin improves pyroptosis in LPs-induced mice mastitis by activating AMPK/Nrf2 and Inhibiting the NF-κB signaling pathway[J]. Oxid Med Cell Longev, 2020, 2020:8845521.
[11]	VéDRINE M, BERTHAULT C, LEROUX C, et al. Sensing of Escherichia coli and LPS by mammary epithelial cells is modulated by O-antigen chain and CD14[J]. PLoS One, 2018, 13(8):e0202664.
[12]	WU Y J, SUN Y W, DONG X W, et al. The synergism of PGN, LTA and LPS in inducing transcriptome changes, inflammatory responses and a decrease in lactation as well as the associated epigenetic mechanisms in bovine mammary epithelial cells[J]. Toxins (Basel), 2020, 12(6):387.
[13]	LIU M, FANG G, YIN S, et al. Caffeic acid prevented LPS-induced injury of primary bovine mammary epithelial cells through inhibiting NF-κB and MAPK activation[J]. Mediators Inflamm, 2019, 2019:1897820.
[14]	LAN R X, ZHAO Z H, LI S Q, et al. Sodium butyrate as an effective feed additive to improve performance, liver function, and meat quality in broilers under hot climatic conditions[J]. Poult Sci, 2020, 99(11):5491-5500.
[15]	OHIRA H, FUJIOKA Y, KATAGIRI C, et al. Butyrate attenuates inflammation and lipolysis generated by the interaction of adipocytes and macrophages[J]. J Atheroscler Thromb, 2013, 20(5):425-442.
[16]	夏令. 奶牛能量负平衡产生的原因、危害及解决办法[J]. 现代畜牧科技, 2021(2):58-59.XIA L. Causes, harms and solutions of negative energy balance in dairy cows[J]. Modern Animal Husbandry Technology, 2021, (2):58-59. (in Chinese)
[17]	邢媛媛, 李大彪, 塔娜, 等. 丁酸对乳腺组织基因表达的调控作用及其机制[J]. 动物营养学报, 2016, 28(10):3064-3069.XING Y Y, LI D B, TA N, et al. Regulatory functions and mechanism of butyrate on gene expressions in mammary glands[J]. Acta Zoonutrimenta Sinica, 2016, 28(10):3064-3069. (in Chinese)
[18]	LIN S C, HARDIE D G. AMPK:sensing glucose as well as cellular energy status[J]. Cell Metab, 2018, 27(2):299-313.
[19]	YAN C H, TIAN X X, LI J Y, et al. A high-fat diet attenuates AMPK α1 in adipocytes to induce exosome shedding and nonalcoholic fatty liver development in vivo[J]. Diabetes, 2021, 70(2):577-588.
[20]	MCCONELL G K. It's well and truly time to stop stating that AMPK regulates glucose uptake and fat oxidation during exercise[J]. Am J Physiol Endocrinol Metab, 2020, 318(4):E564-E567.
[21]	KIM J, YANG G, KIM Y, et al. AMPK activators:mechanisms of action and physiological activities[J]. Exp Mol Med, 2016, 48(4):e224.
[22]	孙梅, 李大彪, 邢媛媛, 等. 氢化可的松对奶牛乳腺上皮细胞乳脂肪合成的影响[J]. 中国细胞生物学学报, 2017, 39(9):1156-1164.SUN M, LI D B, XING Y Y, et al. The effect of hydrocortisone on milk fat synthesis in bovine mammary epithelial cells[J]. Chinese Journal of Cell Biology, 2017, 39(9):1156-1164. (in Chinese)
[23]	SHENG D D, ZHAO S M, GAO L, et al. BabaoDan attenuates high-fat diet-induced non-alcoholic fatty liver disease via activation of AMPK signaling[J]. Cell Biosci, 2019, 9:77.
[24]	POLONI S, BLOM H J, SCHWARTZ I V D. Stearoyl-CoA desaturase-1:Is it the link between sulfur amino acids and lipid metabolism?[J]. Biology (Basel), 2015, 4(2):383-396.
[25]	LEE S M, LEE M H, SON Y K, et al. Omega-3 fatty acid decreases oleic acid by decreasing SCD-1 expression in the liver and kidney of a cyclosporine-induced nephropathy rat model[J]. Ren Fail, 2019, 41(1):211-219.
[26]	IWAMOTO H, ABE M, YANG Y L, et al. Cancer lipid metabolism confers antiangiogenic drug resistance[J]. Cell Metab, 2018, 28(1):104-117.e5.
[27]	LU J C, CHANG Y T, WANG C T, et al. Trichostatin A modulates thiazolidinedione-mediated suppression of tumor necrosis factor α-induced lipolysis in 3T3-L1 adipocytes[J]. PLoS One, 2013, 8(8):e71517.
[28]	XIE L S, ZHANG N, ZHANG Q, et al. Inflammatory factors and amyloid β-induced microglial polarization promote inflammatory crosstalk with astrocytes[J]. Aging (Albany NY), 2020, 12(22):22538-22549.
[29]	SCHWAGER S, DETMAR M. Inflammation and lymphatic function[J]. Front Immunol, 2019, 10:308.
[30]	LIN Y H, LIN Y J, CHANG T H, et al. Pipoxolan suppresses the inflammatory factors of NF-κB, AP-1, and STATs, but activates the antioxidative factor Nrf2 in LPS-stimulated RAW 264. 7 murine macrophage cells[J]. Environ Toxicol, 2020, 35(12):1352-1363.
[31]	CHEN X, LI X, ZHANG W Y, et al. Activation of AMPK inhibits inflammatory response during hypoxia and reoxygenation through modulating JNK-mediated NF-κB pathway[J]. Metabolism, 2018, 83:256-270.

The Mechanism of Sodium Butyrate through AMPK Pathway to Regulate Lipid Metabolism Disorder Caused by LPS in Bovine Mammary Epithelial Cells

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