白消安致小鼠睾丸损伤的机理及其调控通路的研究进展

doi:10.16431/j.cnki.1671-7236.2023.07.021

摘要/Abstract

摘要： 白消安因其具有较强精子毒害作用常被用于制备精原干细胞移植受体，但具体毒性机制尚不清楚，影响了其科学使用及效果提升。研究表明，白消安可引起生精细胞产生氧化应激、炎症反应，并抑制细胞自噬、损伤血睾屏障，毒害睾丸生精机能，但其相互之间的关系及调控通路尚不明确。基于核转录因子κB （nuclear transportation factor κB，NF-κB）通路在炎症反应中的重要调控作用，笔者综述了白消安引起的氧化应激、炎症反应，并损伤血睾屏障及抑制睾丸细胞自噬的研究结果，分析了NF-κB信号通路在白消安毒害睾丸中可能发挥的调控作用，以期深入揭示白消安毒害精子发生的分子机理，为科学使用白消安和避免环境毒素伤害提供参考。

关键词: 白消安; 睾丸; 氧化应激; 炎症反应; 血睾屏障; 自噬

Abstract: Because of its strong toxic effect on spermatogonium, busulfan is often used to prepare spermatogonium stem cell transplantation recipients, but the specific toxic mechanism is still unclear, which affects its scientific use and effect improvement.Studies have shown that busulfan can cause oxidative stress and inflammatory response of spermatogenic cells, inhibit autophagy, damage blood-testis barrier, and poison spermatogenic function of testis, but the relationship between them and the regulatory pathway remain unclear.Based on the important regulatory role of nuclear transportation factor κB(NF-κB) pathway in inflammatory response, the author reviewed the research results of oxidative stress, inflammatory response, damage of blood-testis barrier and inhibition of testicular cell autophagy caused by busulfan, and analyzed the possible regulatory role of NF-κB signaling pathway in testicular toxicity of busulfan.The aim of this study was to reveal the toxic mechanism of busulfan to spermatogenesis, and to provide reference for scientific use of busulfan and to avoid environmental toxins.

Key words: busulfan; testis; oxidative stress; inflammatory reaction; blood-testis barrier; autophagy

中图分类号:

S814.1

吴开慧, 周成利, 赵羚均, 许师源, 刘松奇, 董智豪, 原开敏, 王栋. 白消安致小鼠睾丸损伤的机理及其调控通路的研究进展[J]. 中国畜牧兽医, 2023, 50(7): 2811-2819.

WU Kaihui, ZHOU Chengli, ZHAO Lingjun, XU Shiyuan, LIU Songqi, DONG Zhihao, YUAN Kaimin, WANG Dong. Research Progress on the Mechanism and Regulatory Pathway of Testicular Injury Induced by Busulfan in Mice[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(7): 2811-2819.

参考文献

[1] 陈冬玲, 王得志, 王焱林.白消安对雄性青春期小鼠的生殖系统发育的影响[J].武汉大学学报(医学版), 2016, 37(5):828-832. CHEN D L, WANG D Z, WANG Y L.Effects of busulfan on reproductive system development of male puberty mouse[J].Geomatics of Wuhan University (Medical Edition), 2016, 37(5):828-832.(in Chinese)
[2] MOGHADAM M T, DADFAR R, KHORSANDI L.The effects of ozone and melatonin on busulfan-induced testicular damage in mice[J].JBRA Assisted Reproduction, 2021, 25(2):176-184.
[3] QU N, TERAYAMA H, HIRAYANAGI Y, et al.Induction of experimental autoimmune orchitis by immunization with xenogenic testicular germ cells in mice[J].Journal of Reproductive Immunology, 2017, 121:11-16.
[4] YI W, XIANG-LIANG T, YU Z, et al.DEHP exposure destroys blood-testis barrier (BTB) integrity of immature testes through excessive ROS-mediated autophagy[J].Genes & Diseases, 2018, 5(3):263-274.
[5] ZHANG X, WANG T, DENG T, et al.Damaged spermatogenic cells induce inflammatory gene expression in mouse Sertoli cells through the activation of Toll-like receptors 2 and 4[J].Molecular and Cellular Endocrinology, 2013, 365(2):162-173.
[6] LU J, LIU Z, SHU M, et al.Human placental mesenchymal stem cells ameliorate chemotherapy-induced damage in the testis by reducing apoptosis/oxidative stress and promoting autophagy[J].Stem Cell Research & Therapy, 2021, 12(1):199.
[7] PHAYDEN M S, GHOSH S.Shared principles in NF-kappaB signaling[J].Cell, 2008, 132(3):344-362.
[8] HADDAD J J.Redox and oxidant-mediated regulation of apoptosis signaling pathways:Immuno-pharmaco-redox conception of oxidative siege versus cell death commitment[J].International Immunopharmacology, 2004, 4(4):475-493.
[9] MORGAN M J, LIU Z G.Crosstalk of reactive oxygen species and NF-kappaB signaling[J].Cell Research, 2011, 21(1):103-115.
[10] RAHMAN I, MACNEE W.Role of transcription factors in inflammatory lung diseases[J]. Thorax, 1998, 53(7):601-612.
[11] SCHRECK R, ALBERMANN K, BAEUERLE P A.Nuclear factor kappa B:An oxidative stress-responsive transcription factor of eukaryotic cells (a review)[J].Free Radical Research Communications, 1992, 17(4):221-237.
[12] HAYDEN M S, GHOSH S.NF-kappaB in immunobiology[J].Cell Research, 2011, 21(2):223-244.
[13] LIU J, REN L, WEI J, et al.Fine particle matter disrupts the blood-testis barrier by activating TGF-beta3/p38 MAPK pathway and decreasing testosterone secretion in rat[J]. Environmental Toxicology, 2018, 33(7):711-719.
[14] LU Y, LUO B, LI J, et al.Perfluorooctanoic acid disrupts the blood-testis barrier and activates the TNFalpha/p38 MAPK signaling pathway in vivo and in vitro[J].Archives of Toxicology, 2016, 90(4):971-983.
[15] XIA W, WONG E W, MRUK D D, et al.TGF-beta3 and TNFalpha perturb blood-testis barrier (BTB) dynamics by accelerating the clathrin-mediated endocytosis of integral membrane proteins:A new concept of BTB regulation during spermatogenesis[J].Developmental Biology, 2009, 327(1):48-61.
[16] 冯亚静.NF-κB信号通路介导自噬参与PM2.5致雄性生殖损伤的机制[D].郑州:郑州大学, 2020. FENG Y J.Mechanism of NF-κB signaling pathway mediating autophagy involved in male reproductive damage induced by PM2.5[D].Zhengzhou:Zhengzhou University, 2020.(in Chinese)
[17] ZHOU M, XU W, WANG J, et al.Boosting mTOR-dependent autophagy via upstream TLR4-MyD88-MAPK signalling and downstream NF-kappaB pathway quenches intestinal inflammation and oxidative stress injury[J].EBioMedicine, 2018, 35:345-360.
[18] 唐功.活性氧·抗氧化酶及抗氧化剂之间关系的探讨[J].安徽农业科学, 2010, 38(33):18619-18621. TANG G.The discuss on the relationship among reactive oxygen species, antioxidant enzyme and antioxidant[J]. Journal of Anhui Agricultural Sciences, 2010, 38(33):18619-18621.(in Chinese)
[19] AFZALI A, AMIDI F, KORUJI M, et al.Astaxanthin relieves busulfan-induced oxidative apoptosis in cultured human spermatogonial stem cells by activating the Nrf-2/HO-1 pathway[J].Reproductive Sciences, 2022, 29(2):374-394.
[20] NASIMI P, TABANDEH M R, ROOHI S.Busulfan-mediated oxidative stress and genotoxicity decrease in sperm of Satureja Khuzestanica essential oil-administered mice[J].Systems Biology in Reproductive Medicine, 2018, 64(5):348-357.
[21] ROSTAMI A, VAKILI S, KOOHPEYMA F, et al.Ellagic acid effects on testis, sex hormones, oxidative stress, and apoptosis in the relative sterility rat model following busulfan administration[J].BMC Complementary Medicine and Therapies, 2022, 22(1):170.
[22] LI B, HE X, ZHUANG M, et al.Melatonin ameliorates busulfan-induced spermatogonial stem cell oxidative apoptosis in mouse testes[J]. Antioxidants & Redox Signaling, 2018, 28(5):385-400.
[23] PROBIN V, WANG Y, ZHOU D.Busulfan-induced senescence is dependent on ROS production upstream of the MAPK pathway[J].Free Radical Biology & Medicine, 2007, 42(12):1858-1865.
[24] 雷义, 金文达, 陈锋.活性氧与线粒体损伤[J].南华大学学报(医学版), 2006, 5:648-649. LEI Y, JIN W D, CHEN F.Reactive oxygen species and mitochondrial damage[J] Journal of University of South China (Medical Edition), 2006, 5:648-649.(in Chinese)
[25] 徐道绲, 钱伟, 陈喆, 等.PYDDT通过ROS激活p53诱导结肠癌细胞HCT116凋亡机制的研究[J].中华中医药学刊, 2014, 32(5):1077-1080. XU D G, QIAN W, CHEN Z, et al.PYDDT induces apoptosis of HCT116 cells by ROS activation of p53[J]. Chinese Journal of Traditional Chinese Medicine, 2014, 32(5):1077-1080.(in Chinese)
[26] 董雅洁, 高维娟.bcl-2、bax、caspase-3在细胞凋亡中的作用及其关系[J].中国老年学杂志, 2012, 32(21):4828-4830. DONG Y J, GAO W J.The role of bcl-2, bax and caspase-3 in apoptosis and their relationship[J].Chinese Journal of Gerontology, 2012, 32(21):4828-4830.(in Chinese)
[27] ZHANG X, XIA Q, WEI R, et al.Melatonin protects spermatogonia from the stress of chemotherapy and oxidation via eliminating reactive oxidative species[J].Free Radical Biology & Medicine, 2019, 137:74-86.
[28] ABARIKWU S O, NJOKU R C, JOHN I G, et al.Antioxidant and anti-inflammatory protective effects of rutin and kolaviron against busulfan-induced testicular injuries in rats[J].Systems Biology in Reproductive Medicine, 2022, 68(2):151-161.
[29] QU N, KURAMASU M, HIRAYANAGI Y, et al.Gosha-jinki-gan recovers spermatogenesis in mice with busulfan-induced aspermatogenesis[J]. International Journal of Molecular Sciences, 2018, 19(9):2606.
[30] QU N, KURAMASU M, NAGAHORI K, et al.Co-administration of the traditional medicines hachimi-jio-gan and hochu-ekki-to can reverse busulfan-induced aspermatogenesis[J]. International Journal of Molecular Sciences, 2020, 21(5):1716.
[31] GIRIDHARAM S, SRINIVASAN M.Mechanisms of NF-kappaB p65 and strategies for therapeutic manipulation[J].Journal of Inflammation Research, 2018, 11:407-419.
[32] TONG L, WU S.The role of constitutive nitric-oxide synthase in ultraviolet B light-induced nuclear factor κB activity[J].The Journal of Biological Chemistry, 2014, 289(38):26658-26668.
[33] HADDAD J J, LAND S C.Redox signaling-mediated regulation of lipopolysaccharide-induced proinflammatory cytokine biosynthesis in alveolar epithelial cells[J]. Antioxidants & Redox Signaling, 2002, 4(1):179-193.
[34] HADDAD J J, LAND S C.O₂-evoked regulation of HIF-1alpha and NF-kappaB in perinatal lung epithelium requires glutathione biosynthesis[J].Lung Cellular and Molecular Physiology, 2000, 278(3):L492-L503.
[35] RAHMAN I, MULIER B, GILMOUR P S, et al.Oxidant-mediated lung epithelial cell tolerance:The role of intracellular glutathione and nuclear factor-kappaB[J].Biochemical Pharmacology, 2001, 62(6):787-794.
[36] MRUK D D, CHENG C Y.The mammalian blood-testis barrier:Its biology and regulation[J].Endocrine Reviews, 2015, 36(5):564-591.
[37] 李茹意.白消安毒害小鼠睾丸的机理研究[D].长春:吉林农业大学, 2021. LI R Y.Study on the mechanism of busulfan poisoning mice testis[D].Changchun:Jilin Agricultural University, 2021.(in Chinese)
[38] BALDA M S, MATTER K.Transmembrane proteins of tight junctions[J]. Seminars in Cell & Developmental Biology, 2000, 11(4):281-289.
[39] TSUKITA S, FURUSE M, ITOH M.Multifunctional strands in tight junctions[J].Molecular Cell Biology, 2001, 2(4):285-293.
[40] FU J, LUO Y Y, MIAO S Y, et al.Expression patterns of the proteins associated with cell junctions in mouse testes[J]. Acta Academiae Medicinae Sinicae, 2016, 38(2):205-209.
[41] HEISKALA M, PETERSON P A, YANG Y.The roles of claudin superfamily proteins in paracellular transport[J].Traffic, 2001, 2(2):93-98.
[42] IKENOUCHI J, FURUSE M, FURUSE K, et al.Tricellulin constitutes a novel barrier at tricellular contacts of epithelial cells[J].The Journal of Cell Biology, 2005, 171(6):939-945.
[43] COYNE C B, BERGELSON J M.CAR:A virus receptor within the tight junction[J].Advanced Drug Delivery Reviews, 2005, 57(6):869-882.
[44] EBNET K, SUZUKI A, OHNO S, et al.Junctional adhesion molecules (JAMs):More molecules with dual functions?[J].Journal of Cell Science, 2004, 117(1):19-29.
[45] FENG R, ADENIRAN S O, HUANG F, et al.The ameliorative effect of melatonin on LPS-induced Sertoli cells inflammatory and tight junctions damage via suppression of the TLR4/MyD88/NF-κB signaling pathway in newborn calf[J].Theriogenology, 2022, 179:103-116.
[46] MITIC L L, ANDERSON J M.Molecular architecture of tight junctions[J].Annual Review of Physiology, 1998, 60:121-142.
[47] MCCABE M J, FOO C F, DINGER M E, et al.Claudin-11 and occludin are major contributors to Sertoli cell tight junction function, in vitro[J]. Asian Journal of Andrology, 2016, 18(4):620-626.
[48] UMEDA K, IKENOUCHI J, KATAHIRA-TAYAMA S, et al.ZO-1 and ZO-2 independently determine where claudins are polymerized in tight-junction strand formation[J].Cell, 2006, 126(4):741-754.
[49] DECROUY X, GASC J M, POINTIS G, et al.Functional characterization of Cx43 based gap junctions during spermatogenesis[J]. Journal of Cellular Physiology, 2004, 200(1):146-154.
[50] LIM W, MRUK D D, LEE W M, et al.Connexin 43 is critical to maintain the homeostasis of the blood-testis barrier via its effects on tight junction reassembly[J]. Proceedings of the National Academy of Sciences of the United States of America, 2010, 107(42):17998-18003.
[51] MULLER L, HATZFELD M, KEIL R.Desmosomes as signaling hubs in the regulation of cell behavior[J].Frontiers in Cell and Developmental Biology, 2021, 9:745670.
[52] VLEMINCKX K, KEMLER R.Cadherins and tissue formation:Integrating adhesion and signaling[J].Bioessays, 1999, 21(3):211-220.
[53] XIAO X, CHENG C Y, MRUK D D.Intercellular adhesion molecule-1 is a regulator of blood-testis barrier function[J]. Journal of Cell Science, 2012, 125(23):5677-5689.
[54] ZHAO J, WANG M, WANG Y, et al.Endoplasmic reticulum stress promotes blood-testis barrier impairment in mice with busulfan-induced oligospermia through PERK-eIF2α signaling pathway[J].Toxicology, 2022, 473:153193.
[55] CAI Y, LIU T, FANG F, et al.Involvement of ICAM-1 in impaired spermatogenesis after busulfan treatment in mice[J].Andrologia, 2016, 48(1):37-44.
[56] ALVES M G, RATO L, CARVALHO R A, et al.Hormonal control of Sertoli cell metabolism regulates spermatogenesis[J].Cellular and Molecular Life Sciences, 2013, 70(5):777-793.
[57] LIE P P, CHENG C Y, MRUK D D.Signalling pathways regulating the blood-testis barrier[J].The International Journal of Biochemistry & Cell Biology, 2013, 45(3):621-625.
[58] XIA W, CHENG C Y.TGF-beta3 regulates anchoring junction dynamics in the seminiferous epithelium of the rat testis via the Ras/ERK signaling pathway:An in vivo study[J].Developmental Biology, 2005, 280(2):321-343.
[59] 姚琴, 任明姬, 赵鹏伟.MAPK转导通路在肿瘤发生发展中的作用[J].内蒙古医学杂志, 2014, 46(10):1198-1201. YAO Q, REN M J, ZHAO P W.The role of MAPK transduction pathways in cancer development and progression[J].Inner Mongolia Medical Journal, 2014, 46(10):1198-1201.(in Chinese)
[60] ZHANG H, YIN Y, WANG G, et al.Interleukin-6 disrupts blood-testis barrier through inhibiting protein degradation or activating phosphorylated ERK in Sertoli cells[J].Scientific Reports, 2014, 4:4260.
[61] MIMA M, GREENWALD D, OHLANDER S.Environmental toxins and male fertility[J].Current Urology Reports, 2018, 19(7):50.
[62] LA D K, UPTON P B, SWENBERG J A.Carcinogenic Alkylating Agents[M].Amsterdam:Elsevier, 2010.
[63] ADEGOKE E O, XUE W, MACHEBE N S, et al.Sodium Selenite inhibits mitophagy, downregulation and mislocalization of blood-testis barrier proteins of bovine Sertoli cell exposed to microcystin-leucine arginine (MC-LR) via TLR4/NF-kB and mitochondrial signaling pathways blockage[J]. Ecotoxicology and Environmental Safety, 2018, 166:165-175.
[64] PAN Y, LIU Y, WANG L, et al.MKP-1 attenuates LPS-induced blood-testis barrier dysfunction and inflammatory response through p38 and IκBα pathways[J].Oncotarget, 2016, 7(51):84907-84923.
[65] KLIONSKY D J.The molecular machinery of autophagy:Unanswered questions[J].Journal of Cell Science, 2005, 118(1):7-18.
[66] XILOURI M, STEFANIS L.Chaperone mediated autophagy in aging:Starve to prosper[J].Ageing Research Reviews, 2016, 32:13-21.
[67] 刘杉珊, 李薇.自噬现象及其分子机制的研究进展[J].中外健康文摘, 2011, 8(8):448-451. LIU S S, LI W.Research progress of autophagy and its molecular mechanism[J].Chinese and Foreign Health Abstracts, 2011, 8(8):448-451.(in Chinese)
[68] SIMONSEN A, TOOZE S A.Coordination of membrane events during autophagy by multiple class Ⅲ PI3-kinase complexes[J]. The Journal of Cell Biology, 2009, 186(6):773-782.
[69] 杨喆.多聚嘧啶束结合蛋白1对小鼠血睾屏障完整性和支持细胞自噬的影响[D].贵阳:贵州医科大学, 2021. YANG Z.Effect of polypyrimidine bundle binding protein 1 on the integrity of blood testis barrier and sertoli cell autophagy in mice[D].Guiyang:Guizhou Medical University, 2021.(in Chinese)
[70] MIZUSHIMA N, YOSHIMORI T.How to interpret LC3 immunoblotting[J].Autophagy, 2007, 3(6):542-545.
[71] FUJIOKA Y, NODA N N.Biomolecular condensates in autophagy regulation[J]. Current Opinion in Cell Biology, 2021, 69:23-29.
[72] BENYOUNES A, TAJEDDINE N, TAILLER M, et al.A fluorescence-microscopic and cytofluorometric system for monitoring the turnover of the autophagic substrate p62/SQSTM1[J].Autophagy, 2011, 7(8):883-891.
[73] 马泰, 孙国平, 李家斌.细胞自噬的研究方法[J].生物化学与生物物理进展, 2012, 39(3):204-209. MA T, SUN G P, LI J B.Research methods of autophagy[J].Progress in Biochemistry and Biophysics, 2012, 39(3):204-209.(in Chinese)
[74] VISHNUPRIYA S, PRIYA DHARSHINI L C, SAKTHIVEL K M, et al.Autophagy markers as mediators of lung injury-implication for therapeutic intervention[J]. Life Sciences, 2020, 260:118308.
[75] SARKAR C, ZHAO Z, AUNGST S, et al.Impaired autophagy flux is associated with neuronal cell death after traumatic brain injury[J].Autophagy, 2014, 10(12):2208-2222.
[76] SHINTANI T, KLIONSKY D J.Autophagy in health and disease:A double-edged sword[J].Science, 2004, 306(5698):990-995.
[77] VERMA N, MANNA S K.Advanced glycation end products (AGE) potentiates cell death in p53 negative cells via upregulaion of NF-kappa B and impairment of autophagy[J].Journal of Cellular Physiology, 2017, 232(12):3598-3610.
[78] WEI R, ZHANG X, CAI Y, et al.Busulfan suppresses autophagy in mouse spermatogonial progenitor cells via mTOR of AKT and p53 signaling pathways[J]. Stem Cell Reviews and Reports, 2020, 16(6):1242-1255.