RESEARCH PAPER

Analysis on Effects of Mulberry Leaves at Two Maturities on Fat Body Proteome in Silkworm Based on Isobaric Tags for Relative and Absolute Quantitation Technology

  • XIE Yan ,
  • WANG Hui ,
  • LI Na ,
  • JIA Manli ,
  • HUANG Lu ,
  • SUN Zhichao ,
  • LI Jisheng
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  • Applied Technology R&D Center of Sericulture and Specialty of Hebei Universities, Institute of Sericulture of Chengde Medical University, Chengde 067000, China

Received date: 2021-10-10

  Online published: 2022-05-14

Abstract

This research was to explore the micromechanism of silkworm body adapting to the changes of nutrients in mulberry leaves. The contents of nutrients and active components of tender and aged mulberry leaves, these components including moisture, crude protein, crude fat, soluble sugar, reduced sugar, polysaccharide, total flavonoid, total polyphone and 1-deoxynojirimycin (1-DNJ), were detected and compared. The proteomic profile of fat body from the 5th instar silkworm fed for 3 days with tender and aged mulberry leaves were obtained using isobaric tags for relative and absolute quantitation (iTRAQ) technology, the differentially expressed proteins were also screened and analyzed by bioinformatics method. The results showed that the contents of crude protein, crude fat, reduced sugar, total flavonoid, total polyphone and 1-DNJ tender mulberry leaves were extremely significantly higher than those in aged mulberry leaves (P<0.01). The total of 2 172 fat body proteins were detected by iTRAQ technology, and there were 173 differentially expressed proteins between two groups. Compared with the aged mulberry leaves group, the tender mulberry leaves group had 88 down-regulated expressed proteins and 85 up-regulated expressed proteins. For 111 differentially expressed proteins[fold change (FC)>1.25 or <0.80] of them were used for functional enrichment analysis. GO and KEGG pathway annotation showed these proteins were mainly involved in the biological processes such as substance metabolism, transportation and storage, stress and immune response, body development. Differentially expressed protein interaction analysis found that the ribosomal proteins and neural precursor cell-expressed developmentally down-regulated ptotein 8 (NEDD8) protein had high centrality in interaction network. In conclusion, the maturity of mulberry leaves significantly affects its nutrient contents, resulting the sensitive and extensive differential responses of fat body proteome of silkworm fed tender or aged mulberry leaves.

Cite this article

XIE Yan , WANG Hui , LI Na , JIA Manli , HUANG Lu , SUN Zhichao , LI Jisheng . Analysis on Effects of Mulberry Leaves at Two Maturities on Fat Body Proteome in Silkworm Based on Isobaric Tags for Relative and Absolute Quantitation Technology[J]. Chinese Journal of Animal Nutrition, 2022 , 34(5) : 3271 -3285 . DOI: 10.3969/j.issn.1006-267x.2022.05.053

References

[1] 张辉洁, 孙乐娜, 杨承远, 等.用四臂嗅觉仪观测家蚕对非桑科植物的选择行为[J].蚕业科学, 2012, 38(1):74-81. ZHANG H J, SUN L N, YANG C Y, et al.An observation on food selection behavior of silkworm larvae to non-Moraceae plants with four-arm olfactometer[J].Science of Sericulture, 2012, 38(1):74-81.(in Chinese)
[2] DONG H L, ZHANG S X, CHEN Z H, et al.Differences in gut microbiota between silkworms (Bombyx mori) reared on fresh mulberry (Morus alba var. multicaulis) leaves or an artificial diet[J].RSC Advances, 2018, 8(46):26188-26200.  
[3] ZHOU Z H, YANG H J, CHEN M, et al.Comparative proteomic analysis between the domesticated silkworm (Bombyx mori) reared on fresh mulberry leaves and on artificial diet[J].Journal of Proteome Research, 2008, 7(12):5103-5111.  
[4] 何金涛, 占鹏飞, 郝志华, 等.家蚕人工饲料研究进展[J].蚕桑通报, 2020, 51(1):16-19. HE J T, ZHAN P F, HAO Z H, et al.Research progress of artificial silkworm feed[J].Bulletin of Sericulture, 2020, 51(1):16-19.(in Chinese)
[5] 程洋, 林小兵, 夏冬梅, 等.我国家蚕人工饲料及适应性品种选育研究进展[J].四川蚕业, 2019, 47(2):39-42. CHENG Y, LIN X B, XIA D M, et al.Research progress on the artificial diet and the breeding of varieties suitable for artificial diet of silkworm in China[J].Sichuan Sericulture, 2019, 47(2):39-42.(in Chinese)
[6] 兰凤杰.1-脱氧野尻霉素和脲酶对人工饲料育家蚕代谢的影响及消化酶基因表达研究[D].硕士学位论文.泰安:山东农业大学, 2018. LAN F J.Effects of 1-deoxyoxykomycin and urease on metabolism of silkworm, Bombyx mori, fed by artificial diet and gene expression of digestive enzymes[D].Master's Thesis.Tai'an:Shandong Agricultural University, 2018.(in Chinese)
[7] SABHAT A, MALIL F A, MALIK M A, et al.A comparative study on nutritional composition of some mulberry varieties in relation to expression of economic characters of silkworm races[J].The Journal of Experimental Zoology, 2016, 19(2):935-941.
[8] KERENHAP W, KUMAR V, THIAGARAJAN V.Nutrition quality of mulberry varieties assessed through chemo-assay study of internal tissues and organs of Bombyx mori Linn.[J].Journal of the Entomological Research Society, 2008, 10(1):37-42.
[9] REDDY Y S, MALIK F A, MAMATHA S S, et al.Lipid metabolic profile interrelationships between the silkworm (Bombyx mori L.) and its host plant (Morus) varieties[J].Agricultural Science & Technology, 2009, 10(6):128-134.
[10] ALEBIOSU I B, OLATUNDE G O, PITAN O O R.Variations in some traits of two silkworm Bombyx mori L. (Lepidoptera:Bombycidae) hybrids fed on mulberry leaves of different maturity stages[J].Nigerian Journal of Science, 2013, 47:1-7.
[11] RAHMATHULLA V K, HIMANTHARAJ M T, SRINIVASA G, et al.Association of moisture content in mulberry leaf with nutritional parameters of bivoltine silkworm (Bombyx mori L.)[J].Acta Entomologica Sinica, 2004, 47(6):701-704.
[12] KRISHNAPRASAD N, SANNAPPA B, NAIK B.Effect of feeding mulberry leaves of variable maturity on cocoon traits of Bombyx mori L.[J].Environment and Ecology, 2003, 21(1):112-116.
[13] 陈敏, 宋江波, 李知泉, 等.家蚕作为人类疾病模型与用于药物筛选的研究进展及展望[J].药学学报, 2016, 51(5):690-697. CHEN M, SONG J B, LI Z Q, et al.Progress and perspective of silkworm as a model of human diseases for drug screening[J].Acta Pharmaceutica Sinica, 2016, 51(5):690-697.(in Chinese)
[14] 秦俭, 何宁佳, 向仲怀.家蚕模式化研究进展[J].蚕业科学, 2010, 36(4):645-649. QIN J, HE N J, XIANG Z H.Advances in silkworm modeling research[J].Science of Sericulture, 2010, 36(4):645-649.(in Chinese)
[15] 谢秀枝, 王欣, 刘丽华, 等.iTRAQ技术及其在蛋白质组学中的应用[J].中国生物化学与分子生物学报, 2011, 27(7):616-621. XIE X Z, WANG X, LIU L H, et al.iTRAQ technology and its application in proteomics[J].Chinese Journal of Biochemistry and Molecular Biology, 2011, 27(7):616-621.(in Chinese)
[16] 陈潇飞, 何晓红, 叶绍辉, 等.iTRAQ技术及其在动物蛋白质组学中的研究进展[J].黑龙江畜牧兽医, 2016(7):57-60. CHEN X F, HE X H, YE S H, et al.iTRAQ technology and its research advances in animal proteomics[J].Heilongjiang Animal Science and Veterinary Medicine, 2016(7):57-60.(in Chinese)
[17] 何静, 曾晓英, 岳宣, 等.5龄期不同给桑量对贵蚕1号原蚕生长及经济性状的影响[J].贵州农业科学, 2015, 43(1):114-116. HE J, ZENG X Y, YUE X, et al.Effects of different supplying leaves weight on growth and economic traits of 5th instar silkworm[J].Guizhou Agricultural Sciences, 2015, 43(1):114-116.(in Chinese)
[18] 李瑞, 桑前.家蚕壮蚕期所用桑叶品质与若干经济性状的关系[J].蚕业科学, 1984(4):197-201. LI R, SANG Q.The relationship between mulberry leaf quality and some economic characters of grown silkworm[J].Science of Sericulture, 1984(4):197-201.(in Chinese)
[19] 胡仕叶, 张安臣, 何静, 等.桑叶成熟度对洞庭原蚕生命力及F1卵质的影响[J].贵州农业科学, 2009, 37(11):139-141, 147. HU S Y, ZHANG A C, HE J, et al.Effects of mulberry leaf maturity degree on vital energies and F1 egg quality of Dongting silkworm[J].Guizhou Agricultural Sciences, 2009, 37(11):139-141, 147.(in Chinese)
[20] 侯勇, 赵萍, 刘鸿丽, 等.家蚕脂肪体蛋白质组学研究[J].生物工程学报, 2007, 23(5):867-872. HOU Y, ZHAO P, LIU H L, et al.Proteomics analysis of fat body from silkworm (Bombyx mori)[J].Chinese Journal of Biotechnology, 2007, 23(5):867-872.(in Chinese)
[21] 董久鸣.家蚕 (Bombyx mori) 5龄幼虫脂肪体蛋白质组及其相关功能基因的研究[D].硕士学位论文.杭州:浙江大学, 2008. DONG J M.Proteomic and related functional genes research of fat body from 5 instars silkworm (Bombyx mori)[D].Master's Thesis.Hangzhou:Zhejiang University, 2008.(in Chinese)
[22] CHEN H Q, LI Y J, CHEN K P, et al.Comparative proteomic analysis of Bombyx mori hemolymph and fat body after calorie restriction[J].Acta Biochimica Polonica, 2010, 57(4):505-511.
[23] 田江海.添食TiO2 NPs对家蚕脂肪体生理功能的影响[D].硕士学位论文.苏州:苏州大学, 2017. TIAN J H.Effects of dietary TiO2 NPs on fat body physiological function of Bombyx mori[D].Master's Thesis.Suzhou:Soochow University, 2017.(in Chinese)
[24] SANCHEZ D, LÓPEZ-ARIAS B, TORROJA L, et al.Loss of glial lazarillo, a homolog of apolipoprotein D, reduces lifespan and stress resistance in Drosophila[J].Current Biology, 2006, 16(7):680-686.  
[25] 吕顺霖, 徐俊良, 臧荣春.家蚕幼虫添食蔗糖的效果和机理研究[J].蚕业科学, 1989, 15(1):23-30. LV S L, XU J L, ZANG R C.Study on the effect of sucrose addition on larva of domestic silkworm[J].Acta Sericologica Sinica, 1989, 15(1):23-30.(in Chinese)
[26] 高玉军, 马宝俊, 宋永学, 等.黄芪对家蚕生长影响与其药理的对应关系探讨[J].北方蚕业, 2018, 39(4):14-18. GAO Y J, MA B J, SONG Y X, et al.Effects of Astragalus on silkworm growth and relationship with its pharmacology[J].North Sericulture, 2018, 39(4):14-18.(in Chinese)
[27] CHATTOPADHYAY D, THIRUMURUGAN K.Longevity promoting efficacies of different plant extracts in lower model organisms[J].Mechanisms of Ageing and Development, 2018, 171:47-57.
[28] CHEN C, SONG J B, CHEN M, et al.Rhodiola rosea extends lifespan and improves stress tolerance in silkworm, Bombyx mori[J].Biogerontology, 2016, 17(2):373-381.  
[29] SONG J B, LIU L, HAO K G, et al.Resveratrol elongates the lifespan and improves antioxidant activity in the silkworm Bombyx mori[J].Journal of Pharmaceutical Analysis, 2021, 11(3):374-382.  
[30] 胡乐山, 朱方容.桑叶品质对家蚕产卵数影响的研究[J].广西蚕业, 2000, 37(1):1-7. HU L S, ZHU F R.Effect of the leaf quality of mulberry on eggs-lying of silkworm[J].Guangxi Sericulture, 2000, 37(1):1-7.(in Chinese)
[31] 肖竞军, 吴鹏抟.高温环境中不同成熟度的桑叶对家蚕影响的研究[J].华南农业大学学报, 1987, 8(3):38-46. XIAO J J, WU P T.A study on the influence of the mulberry leaf's maturity of different degrees on the silkworm under high temperature[J].Journal of South China Agricultural University, 1987, 8(3):38-46.(in Chinese)
[32] MILANI M, BECKETT A J, AL-ZEBEEBY A, et al.DRP-1 functions independently of mitochondrial structural perturbations to facilitate BH3 mimetic-mediated apoptosis[J].Cell Death Discovery, 2019, 5:117.
[33] CHICA N, ROZALÉN A E, PÉREZ-HIDALGO L, et al.Nutritional control of cell size by the greatwall-endosulfine-PP2A·B55 pathway[J].Current Biology, 2016, 26(3):319-330.  
[34] 郭璐.Neddylation对小鼠肝细胞凋亡、衰老和脂肪酸β-氧化的影响[D].硕士学位论文.开封:河南大学, 2017. GUO L.The effects of Neddylation on cell apoptosis, cell senescence and β-oxidation of fatty acid in mouse hepatocytes[D].Master's Thesis.Kaifeng:Henan University, 2017.(in Chinese)
[35] TELKOPARAN P, ERKEK S, YAMAN E, et al.Coiled-coil domain containing protein 124 is a novel centrosome and midbody protein that interacts with the Ras-guanine nucleotide exchange factor 1B and is involved in cytokinesis[J].PLoS One, 2013, 8(7):e69289.
[36] UTISPAN K, KOONTONGKAEW S.Macrophage migration inhibitory factor modulates proliferation, cell cycle, and apoptotic activity in head and neck cancer cell lines[J].Journal of Dental Sciences, 2021, 16(1):342-348.  
[37] FANG K, CHU Y, ZHAO Z Y, et al.Enhanced expression of asparagine synthetase under glucose-deprived conditions promotes esophageal squamous cell carcinoma development[J].International Journal of Medical Sciences, 2020, 17(4):510-516.  
[38] CHAPELLIER M, PEÑA-MARTÍNEZ P, RAMAKRISHNAN R, et al.Arrayed molecular barcoding identifies TNFSF13 as a positive regulator of acute myeloid leukemia-initiating cells[J].Haematologica, 2019, 104(10):2006-2016.  
[39] 侯成香, 覃光星, 郭锡杰.家蚕等昆虫贮存蛋白的研究进展[J].中国蚕业, 2016, 37(4):1-5. HOU C X, QIN G X, GUO X J.Advances in studies on storage proteins of silkworm and other insects[J].China Sericulture, 2016, 37(4):1-5.(in Chinese)
[40] 聂作明.家蚕体相关蛋白乙酰化修饰鉴定及其对蚕丝蛋白合成影响的研究[D].博士学位论文.杭州:浙江理工大学, 2015. NIE Z M.Identification of acetylated proteins in silkworm body and its effects on silk protein synthesis[D].Ph.D.Thesis.Hangzhou:Zhejiang Sci-Tech University, 2015.(in Chinese)
[41] ZHU Y N, WANG L Z, LI C C, et al.Artificial selection on storage protein 1 possibly contributes to increase of hatchability during silkworm domestication[J].PLoS Genetics, 2019, 15(1):e1007616.
[42] TOMINAGA T, SHARMA I, FUJITA Y, et al.Myo-inositol oxygenase accentuates renal tubular injury initiated by endoplasmic reticulum stress[J].American Journal of Physiology:Renal Physiology, 2019, 316(2):F301-F315.
[43] GIAMOGANTE F, MARROCCO I, CERVONI L, et al.Punicalagin, an active pomegranate component, is a new inhibitor of PDIA3 reductase activity[J].Biochimie, 2018, 147:122-129.
[44] OH T, KWON M, YU J S, et al.Ent-peniciherqueinone suppresses acetaldehyde-induced cytotoxicity and oxidative stress by inducing ALDH and suppressing MAPK signaling[J].Pharmaceutics, 2020, 12(12):1229.
[45] PEDRO N F, BISELLI J M, MANIGLIA J V, et al.Candidate biomarkers for oral squamous cell carcinoma:differential expression of oxidative stress-related genes[J].Asian Pacific Journal of Cancer Prevention, 2018, 19(5):1343-1349.
[46] BAO A H, MA A Y, ZHANG H, et al.Inducible expression of heat shock protein 20 protects airway epithelial cells against oxidative injury involving the Nrf2-NQO-1 pathway[J].Cell & Bioscience, 2020, 10:120.
[47] DEL CAÑO-ESPINEL M, ACEBES J R, SANCHEZ D, et al.Lazarillo-related Lipocalins confer long-term protection against type Ⅰ Spinocerebellar Ataxia degeneration contributing to optimize selective autophagy[J].Molecular Neurodegeneration, 2015, 10:11.
[48] 薛嘉虹, 朱参战, 胡艳超, 等.ABCG1在肿瘤坏死因子α诱导的氧化应激中的机制研究[J].中国现代医学杂志, 2017, 27(11):14-19. XUE J H, ZHU C Z, HU Y C, et al.ABCG1 inhibits TNF-α-induced oxidative stress by regulating NADPH oxidase and superoxide dismutase[J].China Journal of Modern Medicine, 2017, 27(11):14-19.(in Chinese)
[49] 杨恺清, 赵景霖, 毛小琴.核糖体蛋白的核糖体外功能及其在疾病中的作用[J].生理科学进展, 2021, 52(3):227-230. YANG K Q, ZHAO J L, MAO X Q.The extra-ribosomal functions of ribosomal proteins and their role in disease[J].Progress in Physiological Sciences, 2021, 52(3):227-230.(in Chinese)
[50] VOUSDEN K H, LU X.Live or let die:the cell's response to p53[J].Nature Reviews Cancer, 2002, 2(8):594-604.  
[51] ZHANG Y P, LU H.Signaling to p53:ribosomal proteins find their way[J].Cancer Cell, 2009, 16(5):369-377.  
[52] FRANKLIN D, ZHANG Y P.Nutrient availability dictates the regulation of metabolism by the ribosomal protein-MDM2-p53 pathway[J].Molecular & Cellular Oncology, 2018, 5(3):e1302904.
[53] KIM T H, LESLIE P, ZHANG Y P.Ribosomal proteins as unrevealed caretakers for cellular stress and genomic instability[J].Oncotarget, 2014, 5(4):860-871.  
[54] HE X, LI Y H, DAI M S, et al.Ribosomal protein L4 is a novel regulator of the MDM2-p53 loop[J].Oncotarget, 2016, 7(13):16217-16226.  
[55] BAI D M, ZHANG J F, XIAO W C, et al.Regulation of the HDM2-p53 pathway by ribosomal protein L6 in response to ribosomal stress[J].Nucleic Acids Research, 2014, 42(3):1799-1811.  
[56] ZHU Y H, REN C C, JIANG D Y, et al.RPL34-AS1-induced RPL34 inhibits cervical cancer cell tumorigenesis via the MDM2-P53 pathway[J].Cancer Science, 2021, 112(5):1811-1821.  
[57] VOUSDEN K H, RYAN K M.p53 and metabolism[J].Nature Reviews Cancer, 2009, 9(10):691-700.  
[58] 陈珂, 丁艳平, 王建林, 等.p53参与代谢调控的研究进展[J].生物技术通报, 2016, 32(11):52-58. CHEN K, DING Y P, WANG J L, et al.Research progress on p53-involved metabolic regulation[J].Biotechnology Bulletin, 2016, 32(11):52-58.(in Chinese)
[59] LIU Y, HE Y Z, JIN A W, et al.Ribosomal protein-Mdm2-p53 pathway coordinates nutrient stress with lipid metabolism by regulating MCD and promoting fatty acid oxidation[J].Proceedings of the National Academy of Sciences of the United States of America, 2014, 111(23):E2414-E2422.
[60] NADLER S T, STOEHR J P, SCHUELER K L, et al.The expression of adipogenic genes is decreased in obesity and diabetes mellitus[J].Proceedings of the National Academy of Sciences of the United States of America, 2000, 97(21):11371-11376.  
[61] HAN L, SHEN W J, BITTNER S, et al.PPARs:regulators of metabolism and as therapeutic targets in cardiovascular disease.Part Ⅱ:PPAR-β/δ and PPAR-γ[J].Future Cardiology, 2017, 13(3):279-296.  
[62] MARÉCHAL L, LAVIOLETTE M, RODRIGUE-WAY A, et al.The CD36-PPARγ pathway in metabolic disorders[J].International Journal of Molecular Sciences, 2018, 19(5):1529.
[63] PARK H S, JU U I, PARK J W, et al.PPARγ neddylation essential for adipogenesis is a potential target for treating obesity[J].Cell Death and Differentiation, 2016, 23(8):1296-1311.  
[64] 于洁.家蚕NEDD8功能及其Neddylation修饰蛋白的鉴定[D].硕士学位论文.镇江:江苏科技大学, 2017. YU J.Functional analysis of NEDD8 and identification of its modified proteins in silkworm, Bombyx mori[D].Master's Thesis.Zhenjiang:Jiangsu University of Science and Technology, 2017.(in Chinese)
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