Abstract：Excessive lipid deposition in grass carp Ctenopharyngodon idella adipose tissue as one of the common and prominent problems in aquaculture production can be dealt with induction of adipocyte apoptosis.To investigate the relationship between mitophagy and apoptosis in grass carp adipocytes, 2 trails were designed and conducted in vitro and in vivo. In vitro, juvenile grass carp with body weight of some 20 g were reared in net cages and fed diets containing 0 mg/kg (control group), 10 mg/kg, 20 mg/kg, and 40 mg/kg mitophagy activatorcarbonyl cyanide-3-chlorophenylhydrazone (CCCP) at water temperature of 24—30 ℃for 8 weeks. In vitro trail, the isolated primary adipocytes of grass carpwere cultured, differentiated and matured, and then treated with CCCP at dose of 0(control group), and 20 μmol/L for 24 h.The results in the vivo assay showed that compared with the control group, the treatment of 40 mg/kg of mitophagy activator led to significantly lower grass carp intraperitoneal fat index (P<0.05), and the content of triglycerides (P<0.05), and to significantly elevated the mRNA levels of autophagy-related genes pink1, parkin, atg12, atg5 and apoptosis-related genes caspase3 and caspase9 (P<0.05), without significant change in the size of adipocyte area in each treatment group compared with the control group (P>0.05). In vivo results showed that there were significantly elevated cellular mitophagy-related genes pink1, parkin, atg4, atg7, and atg12(P<0.01), significant increase in the mRNA levels of the apoptosis-related genes bax, caspase9, and the bax/bcl-2 ratio (P<0.05 or P<0.01), and significant decrease in the mRNA of the anti-apoptosis gene bcl-2level (P<0.05), and cellular triglyceride content (P<0.05) in the fish fed the diet containing the activator. It was found that mitophagy activator treatment activated mitophagy and apoptosis. The findings indicate that reduction of intraperitoneal fat index in grass carp may be realized by inducing apoptosis of grass carp adipocytes, thus reducing lipid accumulation in grass carp. Combined in vivo and in vitro assays showed that mitophagy activator treatment activated mitophagy mediated by Pink1/Parkin.
曾彩虹, 边晨晨, 孙健, 吉红, 李汉东, 胡泽超. 激活线粒体自噬对草鱼脂肪细胞凋亡和生长发育的影响[J]. 水产科学, 2025, 44(2): 171-180.
ZENG Caihong, BIAN Chenchen, SUN Jian, JI Hong, LI Handong, HU Zechao. Effects of Activation of Mitophagy on Adipocyte Apoptosis and Growth and Development in Grass Carp Ctenopharyngodon idella. Fisheries Science, 2025, 44(2): 171-180.
[1] 农业农村部渔业渔政管理局,全国水产技术推广总站,中国水产学会. 2024中国渔业统计年鉴[M] .北京:中国农业出版社,2024. [2] TIAN J J, JIN Y Q, YU E M, et al. Farnesoid X receptor is an effective target for modulating lipid accumulation in grass carp, Ctenopharyngodon idella[J] . Aquaculture,2021,534:736248. [3] ELMORE S. Apoptosis:a review of programmed cell death[J] . Toxicologic Pathology,2007,35(4):495-516. [4] GREEN D R, GALLUZZI L, KROEMER G. Mitochondria and the autophagy-inflammation-cell death axis in organismal aging[J] . Science,2011,333(6046):1109-1112. [5] OSELLAME L D, BLACKER T S, DUCHEN M R. Cellular and molecular mechanisms of mitochondrial function[J] . Best Practice & Research Clinical Endocrinology & Metabolism,2012,26(6):711-723. [6] FIVENSON E M, LAUTRUP S, SUN N, et al. Mitophagy in neurodegeneration and aging[J] . Neurochemistry International,2017,109:202-209. [7] SAITO T, SADOSHIMA J. Molecular mechanisms of mitochondrial autophagy/mitophagy in the heart[J] . Circulation Research,2015,116(8):1477-1490. [8] ZHAO C L, HE R Z, SHEN M, et al. PINK1/parkin-mediated mitophagy regulation by reactive oxygen species alleviates rocaglamide A-induced apoptosis in pancreatic cancer cells[J] . Frontiers in Pharmacology,2019,10:968. [9] ZHANG Y L, CHEN L W, LUO Y N, et al. Pink1/parkin-mediated mitophagy regulated the apoptosis of dendritic cells in sepsis[J] . Inflammation,2022,45(3):1374-1387. [10] DUAN T X, HU T, WU C Y, et al. PINK1/Parkin-mediated mitophagy is involved in NaAsO2-induced apoptosis of human hepatic cells through activation of ERK signaling[J] . Toxicology in Vitro,2020,66:104857. [11] DING Q, XIE X L, WANG M M, et al. The role of the apoptosis-related protein BCL-B in the regulation of mitophagy in hepatic stellate cells during the regression of liver fibrosis[J] . Experimental & Molecular Medicine,2019,51(1):1-13. [12] CHEN Y, CHEN H N, WANG K, et al. Ketoconazole exacerbates mitophagy to induce apoptosis by downregulating cyclooxygenase-2 in hepatocellular carcinoma[J] . Journal of Hepatology,2019,70(1):66-77. [13] GUMP J M, THORBURN A. Autophagy and apoptosis:what is the connection?[J] . Trends in Cell Biology,2011,21(7):387-392. [14] LUO L P, LIU M L. Adipose tissue in control of metabolism[J] . Journal of Endocrinology,2016,231(3):R77-R99. [15] RAYALAM S, DELLA-FERA M A, BAILE C A. Phytochemicals and regulation of the adipocyte life cycle[J] . The Journal of Nutritional Biochemistry,2008,19(11):717-726. [16] 邢君霞,吉红,李汉东,等.日粮中裂殖壶藻油配伍外源性EPA对草鱼幼鱼脂肪酸组成、FAD、ELO基因表达及脂质代谢的影响[J] .水产学报,2021,45(10):1642-1656. [17] JIN A, LEI C X, TIAN J J, et al. Dietary docosahexaenoic acid decreased lipid accumulation via inducing adipocytes apoptosis of grass carp, Ctenopharygodon idella[J] . Fish Physiology and Biochemistry,2018,44(1):197-207. [18] LI H D, JI S H, YUAN X T, et al. Eicosapentaenoic acid (EPA) improves grass carp (Ctenopharyngodonidellus) muscle development and nutritive value by activating the mTOR signaling pathway[J] . Fish Physiology and Biochemistry,2024,50(2):687-703. [19] 陈永平,韩现芹,陈建,等.MS-222在草鱼、花鲈、鲫鱼组织中的富集与消除[J] .水产科学,2019,38(4):527-532. [20] BIAN C C, YUAN X T, ZENG C H, et al. Docosahexaenoic acid (DHA) inhibits abdominal fat accumulation by promoting adipocyte apoptosis through PPARγ-LC3-BNIP3 pathway-mediated mitophagy[J] . Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids,2024,1869(1):159425. [21] 姜鹏,骆丽婷,李胜杰,等.草鱼肠系膜脂肪组织高质量总RNA提取方法的研究[J] .水产科学,2021,40(2):166-171. [22] SHI X C, JIN A, SUN J, et al. α-lipoic acid ameliorates n-3 highly-unsaturated fatty acids induced lipid peroxidation via regulating antioxidant defenses in grass carp (Ctenopharyngodonidellus)[J] . Fish & Shellfish Immunology,2017,67:359-367. [23] 胡泽超,邹孝翠,孙健,等.草鱼脂肪组织及脂肪细胞qRT-PCR内参基因的筛选[J] .水产科学,2023,42(4):547-555. [24] YUAN X C, LIANG X F, LIU L W, et al. Fat deposition pattern and mechanism in response to dietary lipid levels in grass carp, Ctenopharyngodonidellus[J] . Fish Physiology and Biochemistry,2016,42(6):1557-1569. [25] LIU P, JI H, LI C, et al. Ontogenetic development of adipose tissue in grass carp (Ctenopharyngodonidellus)[J] . Fish Physiology and Biochemistry,2015,41(4):867-878. [26] LUO Y, HU C T, QIAO F, et al. Gemfibrozil improves lipid metabolism in Nile tilapia Oreochromis niloticus fed a high-carbohydrate diet through peroxisome proliferator activated receptor-α activation[J] . General and Comparative Endocrinology,2020,296:113537. [27] LU R H, LIANG X F, WANG M, et al. The role of leptin in lipid metabolism in fatty degenerated hepatocytes of the grass carp Ctenopharyngodonidellus[J] . Fish Physiology and Biochemistry,2012,38(6):1759-1774. [28] HUANG X C, BIAN C C, JI H, et al. DHA induces adipocyte lipolysis through endoplasmic reticulum stress and the cAMP/PKA signaling pathway in grass carp (Ctenopharyngodon idella)[J] . Animal Nutrition,2023,13:185-196. [29] TODORČEVIĆ M, HODSON L. The effect of marine derived n-3 fatty acids on adipose tissue metabolism and function[J] . Journal of Clinical Medicine,2015,5(1):3. [30] TIAN J J, LEI C X, JI H, et al. Dietary arachidonic acid has a time-dependent differential impact on adipogenesis modulated via COX and LOX pathways in grass carp Ctenopharyngodonidellus[J] . Lipids,2016,51(12):1325-1338. [31] XU Y C, XU Y H, ZHAO T, et al. Waterborne Cu exposure increased lipid deposition and lipogenesis by affecting Wnt/β-catenin pathway and the β-catenin acetylation levels of grass carp Ctenopharyngodon idella[J] . Environmental Pollution,2020,263:114420. [32] WANDEROY S, HEES J T, KLESSE R, et al. Kill one or kill the many:interplay between mitophagy and apoptosis[J] . Biological Chemistry,2020,402(1):73-88. [33] CHANG G R, CHIU Y S, WU Y Y, et al. Rapamycin protects against high fat diet–induced obesity in C57BL/6J mice[J] . Journal of Pharmacological Sciences,2009,109(4):496-503. [34] HAN S L, WANG J, LI L Y, et al. The regulation of rapamycin on nutrient metabolism in Nile tilapia fed with high-energy diet[J] . Aquaculture,2020,520:734975. [35] EL-CHAÂR D, GAGNON A, SORISKY A. Inhibition of insulin signaling and adipogenesis by rapamycin:effect on phosphorylation of p70 S6 kinase vs eIF4E-BP1[J] . International Journal of Obesity and Related Metabolic Disorders,2004,28(2):191-198. [36] SOLIMAN G A, ACOSTA-JAQUEZ H A, FINGAR D C. mTORC1 inhibition via rapamycin promotes triacylglycerol lipolysis and release of free fatty acids in 3T3-L1 adipocytes[J] . Lipids,2010,45(12):1089-1100. [37] HAN S L, WANG J, ZHANG Y X, et al. Inhibited autophagy impairs systemic nutrient metabolism in Nile tilapia[J] . Comparative Biochemistry and Physiology Part A:Molecular & Integrative Physiology,2019,236:110521. [38] CHANG Y Y, JUHÁSZ G, GORAKSHA-HICKS P, et al. Nutrient-dependent regulation of autophagy through the target of rapamycin pathway[J] . Biochemical Society Transactions,2009,37(Pt 1):232-236. [39] GUO R, ZHANG Y M, TURDI S, et al. Adiponectin knockout accentuates high fat diet-induced obesity and cardiac dysfunction:role of autophagy[J] . Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease,2013,1832(8):1136-1148. [40] GAO G Y, CHEN W W, YAN M J, et al. Rapamycin regulates the balance between cardiomyocyte apoptosis and autophagy in chronic heart failure by inhibiting mTOR signaling[J] . International Journal of Molecular Medicine,2020,45(1):195-209. [41] LI Q, GAO S E, KANG Z R, et al. Rapamycin enhances mitophagy and attenuates apoptosis after spinal ischemia-reperfusion injury[J] . Frontiers in Neuroscience,2018,12:865. [42] LI W, JIANG W S, SU Y R, et al. PINK1/Parkin-mediated mitophagy inhibits osteoblast apoptosis induced by advanced oxidation protein products[J] . Cell Death & Disease,2023,14(2):88. [43] WANG Q C, HE G, MAI K S, et al. Chronic rapamycin treatment on the nutrient utilization and metabolism of juvenile turbot (Psettamaxima)[J] . Scientific Reports,2016,6:28068. [44] PARK Y S, CHOI S E, KOH H C. PGAM5 regulates PINK1/Parkin-mediated mitophagy via DRP1 in CCCP-induced mitochondrial dysfunction[J] . Toxicology Letters,2018,284:120-128. [45] KONCHA R R, RAMACHANDRAN G, SEPURI N B V, et al. CCCP-induced mitochondrial dysfunction - characterization and analysis of integrated stress response to cellular signaling and homeostasis[J] . The FEBS Journal,2021,288(19):5737-5754. [46] SOUTAR M P M, KEMPTHORNE L, ANNUARIO E, et al. FBS/BSA media concentration determines CCCP′s ability to depolarize mitochondria and activate PINK1-PRKN mitophagy[J] . Autophagy,2019,15(11):2002-2011. [47] 陈红光,谢克亮,于泳浩.线粒体自噬的调控分子在不同病生理过程中的作用机制研究进展[J] .中国中西医结合外科杂志,2019,25(5):839-843.
2025, Vol. 44 
