摘要
本研究以小鼠和牦牛为研究对象,首先在小鼠上建立了单核细胞吞噬诱导凋亡卵泡颗粒细胞的模型,然后在此基础上,对牦牛单核细胞吞噬诱导凋亡颗粒细胞过程的形态变化做了深入的研究。
在小鼠上,本实验采用体外培养和诱导凋亡的方式,模拟体内单核细胞吞噬由大蒜诱导凋亡的卵泡颗粒细胞的过程,在不同时点(1h,2h,3h,4h,5h)对共培养物进行瑞氏染色后,用显微镜观察并照相,对吞噬程度随时间变化的规律进行回归分析。结果显示,在颗粒细胞凋亡形变的初期,即可被单核细胞识别,形成吞噬泡,与此同时,颗粒细胞的凋亡程度也越来越大,在吞噬泡内可见到凋亡细胞崩解的碎片。对照片进行分类计数发现,单核细胞与颗粒细胞共同培养约3 h的时候,外接凋亡细胞的单核细胞个数与内吞凋亡细胞的单核细胞个数的比值接近1,说明此时单核细胞中半数呈现吞入状态,半数呈现外接(识别阶段)状态,笔者建议将此状态的时间点称为“半吞期”。SPSS13.0回归分析,可得线性回归方程y=-0.247x+1.644,R~2=0.912,回归方程显著性检验F=31.095,P=0.011(＜0.05),回归系数显著性检验T=-5.576,P=0.011(＜0.05)。
在牦牛上,以先期的小鼠实验为基础,同样模拟了牦牛体内单核细胞吞噬由大蒜诱导凋亡的卵泡颗粒细胞的过程,在不同时点(1h,2h,3h,4h,5h)对共培养物进行瑞氏染色后,用显微镜观察并照相,对吞噬程度随时间变化的规律进行回归分析。同时,对牦牛单核细胞吞噬诱导凋亡的卵泡颗粒细胞的过程,进行了扫描电镜和透射电镜的观察。结果显示,牦牛单核细胞吞噬诱导凋亡卵泡颗粒细胞的光镜下的过程基本与在小鼠上观察到的相同,半吞期为4h。SPSS15回归分析,得线性回归方程y=-0.540x+2.958,R~2=0.937,回归方程显著性检验F=44.766,P=0.007(＜0.01),回归系数显著性检验T=-6.691,P=0.007(＜0.01)。在扫描电镜和透射电镜下,清晰可见颗粒细胞的凋亡和其被吞噬的过程。
综上所述,这种对早期凋亡细胞吞噬过程的体外模拟可以作为一种定量分析的方法,判断吞噬能力的变化,从而可用于对一些影响因素,如促进或抑制吞噬的药物,进行定量分析。以小鼠模型为基础,可在许多的动物上研究其他吞噬细胞的吞噬能力。实验表明小鼠模型在牦牛上是适用的,且得出的半吞期及相关的形态学研究结果,一方面扩大了对牦牛生殖系统凋亡以及凋亡清除的认识,另一方面为深入的研究提供了依据。另外,实验过程中发现,反相处理瑞氏染色的照片,可获得单一荧光染色的效果。
The present study firstly established a method for quantitative measurement of phagocytosis in mouse. Then the further research has been carried out about the morphologic change of phagocytosis in yak.
On the mouse, the phagocytic process of apoptotic granulosa cells by monocytes was imitated in vitro. Monocytes and granulosa cells were isolated from Kunming mice and cultured. Granulosa cells were induced to apoptosis by garlic, and then co-cultured with monocytes. At different time points (1 h, 2 h, 3 h, 4 h, 5 h), co-cultured cells were assessed by Wright's staining. The results showed that at the beginning of morphological changes in apoptotic granulosa cells, monocytes captured the apoptotic cells. Meanwhile, the apoptosis of granulosa cells were continuing. Debris was found in phagocytic vacuole. At the point of 3 h after co-culture, the ratio between the number of monocytes which attached apoptotic granulosa cells and that which engulfed the apoptotic cells was close to one. Namely, half of monocytes were in the state of recognition and half were in a state of engulfment. The author suggested to name it as 'half phagocytic period'. Regression analysis showed that the equation of linear regression was y = -0.247x + 1.644, R~2=0.912, F=31.095, P=0.011 (<0.05), T= -5.576, P=0.011 (<0.05).
On the yak, according to the mode of mouse above, the phagocytic process of apoptotic granulosa cells by monocytes was also imitated in vitro. At different time points (1 h, 2 h, 3 h, 4 h, 5 h), co-cultured cells were assessed by Wright's staining. In addition, the process was observed through scanning electron microscope (SEM) and transmission electron microscope (TEM). The results showed that the morphological changes under microscope were same with that of mouse in the main. At the point of 4 h after co-culture, the ratio between the number of monocytes which attached apoptotic granulosa cells and that which engulfed the apoptotic cells was close to one. Regression analysis showed that the equation of linear regression was y = -0.540x + 2.958, R~2 = 0.937, F=44.766, P=0.007 (<0.01), T= -6.691, P=0.007 (<0.01). Under the SEM and TEM, the apoptosis and phagocytosis were apparent.
In conclusion, the present mode of phagocytosis can be used as a method of quantitative assay, which could analyze some effective factors such as medicines that could enhance or restrain phagocytosis. The mode of mouse fit on the study of yak. The result of half phagocytic period and morphological changes enlarged the understanding of apoptosis and phagocytosis, providing base for further research. Based on the mode, the ability of phagocytosis could be studied on other animals. In addition, it was notable that effect of fluorochrome stain could be got through picture processing of 'Invert' on the pictures.
引文
[1]Han H,Iwanaga T,Uchiyama Y,et al.Aggregation of macrophages in the tips of intestinal villi in guinea pigs:their possible role in the phagocytosis of effete epithelial cells.[J].Cell Tissue Res.1993,271(3):407-416.
[2]Hopkinson-Woolley J,Hughes D,Gordon S,et al.Macrophage recruitment during limb development and wound healing in the embryonic and foetal mouse.[J].J Cell Sci.1994,107(Pt 5):1159-1167.
[3]Cohen J J.Programmed cell death in the immune system.[J].Adv Immunol.1991,50:55-85.
[4]Savill J,Dransfield I,Gregory C,et al.A blast from the past:clearance of apoptotic cells regulates immune responses.[J].Nat Rev Immunol.2002,2(12):965-975.
[5]Dini L,Autuori F,Lentini A,et al.The clearance of apoptotic cells in the liver is mediated by the asialoglycoprotein receptor.[J].FEBS Lett.1992,296(2):174-178.
[6]Dini L,Lentini A,Diez G D,et al.Phagocytosis of apoptotic bodies by liver endothelial cells.[J].J Cell Sci.1995,108(Pt 3):967-973.
[7]Walsh G M,Sexton D W,Blaylock M G,et al.Resting and cytokine-stimulated human small airway epithelial cells recognize and engulf apoptotic eosinophils.[J].Blood.1999,94(8):2827-2835.
[8]Savill J,Smith J,Sarraf C,et al.Glomerular mesangial cells and inflammatory macrophages ingest neutrophils undergoing apoptosis.[J].Kidney Int.1992,42(4):924-936.
[9]Hall S E,Savill J S,Henson P M,et al.Apoptotic neutrophils are phagocytosed by fibroblasts with participation of the fibroblast vitronectin receptor and involvement of a mannose/fucose-specific lectin.[J].J Immunol.1994,153(7):3218-3227.
[10]Thompson C B.Apoptosis in the pathogenesis and treatment of disease.[J].Science.1995,267(5203):1456-1462.
[11]Damasio A R.[Z].New York:Harcourt Brace,1999.
[12]Guerrero I,Ruiz I A A.Development.Longing for ligand:hedgehog,patched,and cell death.[J].Science.2003,301(5634):774-776.
[13]Thibert C,Teillet M A,Lapointe F,et al.Inhibition of neuroepithelial patched-induced apoptosis by sonic hedgehog.[J].Science.2003,301(5634):843-846.
[14]Werlen G,Hausmann B,Naeher D,et al.Signaling life and death in the thymus:timing is everything.[J].Science.2003,299(5614):1859-1863.
[15]Cotran R S,Kumar V,Collins T,et al.[Z].Philadelphia;London:W.B.Saunders,1998.
[16]Li M O,Sarkisian M R,Mehal W Z,et al.Phosphatidylserine receptor is required for clearance of apoptotic cells.[J]. Science. 2003,302(5650): 1560-1563.
[17] Wang X, Wu Y C, Fadok V A, et al. Cell corpse engulfment mediated by C. elegans phosphatidylserine receptor through CED-5 and CED-12.[J]. Science. 2003, 302(5650): 1563-1566.
[18] Savill J, Gregory C, Haslett C. Cell biology. Eat me or die.[J]. Science. 2003, 302(5650): 1516-1517.
[19] Mayer. Immunology — Chapter One: Innate (non-specific) Immunity[Z]. Gene. Microbiology and Immunology On-Line Textbook. USC School of Medicine. : 2006.
[20] Delves P J, Roitt I M. Roitt's Essential Immunology [Z]. Maiden, Mass.: Blackwell Pub., 2006.
[21] Hoffbrand A V, Moss P A H , Pettit J E. Essential Haematology (4th edition)[Z]. Maiden, Mass.; Oxford: Blackwell Pub., 2006.
[22] Little W, Fowler H W, Coulson J S, et al. The Shorter Oxford English Dictionary[Z]. London: Guild Pub., 19831566-1567.
[23] Jr Schmalstieg F C, Goldman A S. Ilya Ilich Metchnikoff (1845-1915) and Paul Ehrlich (1854-1915): the centennial of the 1908 Nobel Prize in Physiology or Medicine.[J]. J Med Biogr. 2008,16(2): 96-103.
[24] Janeway. Chapter: Evolution of the innate immune system.see Bibliography, retrieved on March 20,2009 [Z].
[25] Ernst J. D. And Stendahl O. (Phagocytosis of Bacteria and Bacterial Pathogenicity[Z]. Cambridge University Press: 2006.
[26] Robinson J.p. And Babcock G. F. (Phagocyte Function —A guide for research and clinical evaluation[Z]. Wiley-Liss: 1998.
[27] Janeway. Chapter: Induced innate responses to infection, see Bibliography, retrieved on March 20,2009[Z].
[28] Fang F C. Antimicrobial reactive oxygen and nitrogen species: concepts and controversies.[J]. Nat Rev Microbiol. 2004,2(10): 820-832.
[29] Janeway. Chapter: Antigen Presentation to T Lymphocytes. see Bibliography, retrieved on March 20,2009 [Z].
[30] Hampton M B, Vissers M C, Winterbourn C C. A single assay for measuring the rates of phagocytosis and bacterial killing by neutrophils.[J]. J Leukoc Biol. 1994, 55(2): 147-152.
[31] Sompayrac L. How the Immune System Works (3rd edition)[Z]. Blackwell Publishing: 2008.
[32] Zhou Z, Yu X. Phagosome maturation during the removal of apoptotic cells: receptors lead the way.[J]. Trends Cell Biol. 2008,18(10): 474-485.
[33] Zen K, Parkos C A. Leukocyte-epithelial interactions.[J]. Curr Opin Cell Biol. 2003,15(5):557-564.
[34] Takahashi K, Naito M, Takeya M. Development and heterogeneity of macrophages and their related cells through their differentiation pathways.[J]. Pathol Int. 1996,46(7): 473-485.
[35] Krombach F, Munzing S, Allmeling A M, et al. Cell size of alveolar macrophages: an interspecies comparison.[J]. Environ Health Perspect. 1997,105 Suppl 5: 1261-1263.
[36] Bowers W (Immunology -Chapter Thirteen: Immunoregulation[Z]. Microbiology and Immunology On-Line Textbook. USC School of Medicine..
[37] Stvrtinová V, Jakubovsky J, I H ("Neutrophils, central cells in acute inflammation". Inflammation and Fever from Pathophysiology: Principles of Disease. [Z]. Computing Centre, Slovak Academy of Sciences: Academic Electronic Press.Retrieved on March 28, 2009. .
[38] Linderkamp O, Ruef P, Brenner B, et al. Passive deformability of mature, immature, and active neutrophils in healthy and septicemic neonates.[J]. Pediatr Res. 1998,44(6): 946-950.
[39] Paoletti R. N A A R. Phagocytes: Biology, Physiology, Pathology, and Pharmacotherapeutics[Z]. The New York Academy of Sciences: 1997.
[40] Soehnlein O, Kenne E, Rotzius P, et al. Neutrophil secretion products regulate anti-bacterial activity in monocytes and macrophages.[J]. Clin Exp Immunol. 2008, 151(1): 139-145.
[41] Soehnlein O, Kai-Larsen Y, Frithiof R, et al. Neutrophil primary granule proteins HBP and HNP1-3 boost bacterial phagocytosis by human and murine macrophages.[J]. J Clin Invest. 2008,118(10): 3491-3502.
[42] Steinman R M, Cohn Z A. Identification of a novel cell type in peripheral lymphoid organs of mice. I. Morphology, quantitation, tissue distribution.[J]. J Exp Med. 1973,137(5):1142-1162.
[43] Steinman, Ralph. "Dendritic Cells"[Z]. The Rockefeller University .Retrieved on November 14, 2008. .
[44] Guermonprez P, Valladeau J, Zitvogel L, et al. Antigen presentation and T cell stimulation by dendritic cells.[J]. Annu Rev Immunol. 2002,20: 621-667.
[45] Sallusto F, Lanzavecchia A. The instructive role of dendritic cells on T-cell responses.[J]. Arthritis Res. 2002,4 Suppl 3: 127-132.
[46] Stelekati E, Orinska Z, Bulfone-Paus S. Mast cells in allergy: innate instructors of adaptive responses.[J]. Immunobiology. 2007,212(6): 505-519.
[47] Malaviya R, Abraham S N. Mast cell modulation of immune responses to bacteria.[J]. Immunol Rev. 2001,179:16-24.
[48] Connell I, Agace W, Klemm P, et al. Type 1 fimbrial expression enhances Escherichia coli virulence for the urinary tract.[J]. Proc Natl Acad Sci U S A. 1996, 93(18): 9827-9832.
[49] Malaviya R, Twesten N J, Ross E A, et al. Mast cells process bacterial Ags through a phagocytic route for class I MHC presentation to T cells.[J]. J Immunol. 1996, 156(4): 1490-1496.
[50] Taylor M L, Metcalfe D D. Mast cells in allergy and host defense.[J]. Allergy Asthma Proc. 2001,22(3): 115-119.
[51] Birge R B, Ucker D S. Innate apoptotic immunity: the calming touch of death.[J]. Cell Death Differ. 2008,15(7): 1096-1102.
[52] Couzinet S, Cejas E, Schittny J, et al. Phagocytic uptake of Encephalitozoon cuniculi by nonprofessional phagocytes.[J]. Infect Immun. 2000,68(12): 6939-6945.
[53] Rabinovitch M. Professional and non-professional phagocytes: an introduction.[J]. Trends Cell Biol. 1995, 5(3): 85-87.
[54] Todar K. "Mechanisms of Bacterial Pathogenicity: Bacterial Defense Against Phagocytes"[Z]. 2008.
[55] Alexander J, Satoskar A R, Russell D G. Leishmania species: models of intracellular parasitism.[J]. J Cell Sci. 1999,112 Pt 18:2993-3002.
[56] Celli J, Finlay B B. Bacterial avoidance of phagocytosis.[J]. Trends Microbiol. 2002, 10(5): 232-237.
[57] Valenick L V, Hsia H C, Schwarzbauer J E. Fibronectin fragmentation promotes alpha4betal integrin-mediated contraction of a fibrin-fibronectin provisional matrix.[J]. Exp Cell Res. 2005, 309(1): 48-55.
[58] Burns S M, Hull S I. Loss of resistance to ingestion and phagocytic killing by O(-) and K(-) mutants of a uropathogenic Escherichia coli O75:K5 strain.[J]. Infect Immun. 1999,67(8): 3757-3762.
[59] Vuong C, Kocianova S, Voyich J M, et al. A crucial role for exopolysaccharide modification in bacterial biofilm formation, immune evasion, and virulence.[J]. J Biol Chem. 2004,279(52): 54881-54886.
[60] Melin M, Jarva H, Siira L, et al. Streptococcus pneumoniae capsular serotype 19F is more resistant to C3 deposition and less sensitive to opsonophagocytosis than serotype 6B.[J]. Infect Immun. 2009, 77(2): 676-684.
[61] Foster T J. Immune evasion by staphylococci.[J]. Nat Rev Microbiol. 2005, 3(12): 948-958.
[62] Sansonetti P. Phagocytosis of bacterial pathogens: implications in the host response.[J]. Semin Immunol. 2001,13(6): 381-390.
[63] Dersch P, Isberg R R. A region of the Yersinia pseudotuberculosis invasin protein enhances integrin-mediated uptake into mammalian cells and promotes self-association.[J]. EMBO J. 1999, 18(5): 1199-1213.
[64] Antoine J C, Prina E, Lang T, et al. The biogenesis and properties of the parasitophorous vacuoles that harbour Leishmania in murine macrophages.[J]. Trends Microbiol. 1998,6(10):392-401.
[65] Masek K S, C A H (Eurekah Bioscience Collection: Evasion of Phagosome Lysosome Fusion and Establishment of a Replicative Organelle by the Intracellular Pathogen Legionella pneumophila. Landes Bioscience. [Z].
[66] Das D, Saha S S, Bishayi B. Intracellular survival of Staphylococcus aureus: correlating production of catalase and superoxide dismutase with levels of inflammatory cytokines.[J]. Inflamm Res. 2008, 57(7): 340-349.
[67] Hara H, Kawamura I, Nomura T, et al. Cytolysin-dependent escape of the bacterium from the phagosome is required but not sufficient for induction of the Th1 immune response against Listeria monocytogenes infection: distinct role of Listeriolysin O determined by cytolysin gene replacement.[J]. Infect Immun. 2007,75(8): 3791-3801.
[68] Datta V, Myskowski S M, Kwinn L A, et al. Mutational analysis of the group A streptococcal operon encoding streptolysin S and its virulence role in invasive infection.[J]. Mol Microbiol. 2005, 56(3): 681-695.
[69] Iwatsuki K, Yamasaki O, Morizane S, et al. Staphylococcal cutaneous infections: invasion, evasion and aggression.[J]. J Dermatol Sci. 2006,42(3): 203-214.
[70] Denkers E Y, Butcher B A. Sabotage and exploitation in macrophages parasitized by intracellular protozoans.[J]. Trends Parasitol. 2005, 21(1): 35-41.
[71] Gregory D J, Olivier M. Subversion of host cell signalling by the protozoan parasite Leishmania.[J]. Parasitology. 2005, 130 Suppl: 27-35.
[72] Heinzelmann M, Mercer-Jones M A, Passmore J C. Neutrophils and renal failure.[J]. Am J Kidney Dis. 1999,34(2): 384-399.
[73] Lee W L, Downey G P. Neutrophil activation and acute lung injury.[J]. Curr Opin Crit Care. 2001,7(1): 1-7.
[74] Moraes T J, Zurawska J H, Downey G P. Neutrophil granule contents in the pathogenesis of lung injury.[J]. Curr Opin Hematol. 2006, 13(1): 21-27.
[75] Abraham E. Neutrophils and acute lung injury.[J]. Crit Care Med. 2003, 31(4 Suppl): 195-199.
[76] Ricevuti G. Host tissue damage by phagocytes.[J]. Ann N Y Acad Sci. 1997,832:426-448.
[77] Charley B, Riffault S, Van Reeth K. Porcine innate and adaptative immune responses to influenza and coronavirus infections.[J]. Ann N Y Acad Sci. 2006, 1081: 130-136.
[78] Paoletti R. N A A R. Phagocytes: Biology, Physiology, Pathology, and Pharmacotherapeutics[Z]. The New York Academy of Sciences: 1997.
[79] Parham P. The Immune System, 2nd edition [Z]. Garland Science.
[80] Ishimoto H, Yanagihara K, Araki N, et al. Single-cell observation of phagocytosis by human blood dendritic cells.[J]. Jpn J Infect Dis. 2008, 61(4): 294-297.
[81] Boettner D R, Huston C D, Linford A S, et al. Entamoeba histolytica phagocytosis of human erythrocytes involves PATMK, a member of the transmembrane kinase family.[J]. PLoS Pathog.2008,4(1):8.
[82] DPDx - Amebiasis[Z]. http://www.dpd.cdc.gov/DPDx/html/Amebiasis.htm.
[83] Gronlien H K, Berg T, Lovlie A M. In the polymorphic ciliate Tetrahymena vorax, the non-selective phagocytosis seen in microstomes changes to a highly selective process in macrostomes.[J]. J Exp Biol. 2002,205(Pt 14): 2089-2097.
[84] Stibor H, Sommer U. Mixotrophy of a photosynthetic flagellate viewed from an optimal foraging perspective.[J]. Protist. 2003,154(1): 91-98.
[85] Hart S P, Dransfield I, Rossi A G. Phagocytosis of apoptotic cells.[J]. Methods. 2008,44(3): 280-285.
[86] Dransfield I, Stocks S C, Haslett C. Regulation of cell adhesion molecule expression and function associated with neutrophil apoptosis.[J]. Blood. 1995, 85(11): 3264-3273.
[87] Savill J S, Wyllie A H, Henson J E, et al. Macrophage phagocytosis of aging neutrophils in inflammation. Programmed cell death in the neutrophil leads to its recognition by macrophages.[J]. J Clin Invest. 1989, 83(3): 865-875.
[88] Stern M, Savill J, Haslett C. Human monocyte-derived macrophage phagocytosis of senescent eosinophils undergoing apoptosis. Mediation by alpha v beta 3/CD36/thrombospondin recognition mechanism and lack of phlogistic response.[J]. Am J Pathol. 1996,149(3): 911-921.
[89] Fadok V A, Bratton D L, Konowal A, et al. Macrophages that have ingested apoptotic cells in vitro inhibit proinflammatory cytokine production through autocrine/paracrine mechanisms involving TGF-beta, PGE2, and PAF.[J]. J Clin Invest. 1998, 101(4): 890-898.
[90] Woolley K L, Gibson P G, Carty K, et al. Eosinophil apoptosis and the resolution of airway inflammation in asthma.[J]. Am J Respir Crit Care Med. 1996, 154(1): 237-243.
[91] Kotecha S, Mildner R J, Prince L R, et al. The role of neutrophil apoptosis in the resolution of acute lung injury in newborn infants.[J]. Thorax. 2003, 58(11): 961-967.
[92] Savill J, Dransfield I, Hogg N, et al. Vitronectin receptor-mediated phagocytosis of cells undergoing apoptosis.[J].Nature. 1990,343(6254): 170-173.
[93] Albert M L, Kim J I, Birge R B. alphavbeta5 integrin recruits the CrkII-Dock180-rac1 complex for phagocytosis of apoptotic cells.[J]. Nat Cell Biol. 2000,2(12): 899-905.
[94] Savill J, Hogg N, Ren Y, et al. Thrombospondin cooperates with CD36 and the vitronectin receptor in macrophage recognition of neutrophils undergoing apoptosis.[J]. J Clin Invest. 1992,90(4): 1513-1522.
[95] Duvall E, Wyllie A H, Morris R G. Macrophage recognition of cells undergoing programmed cell death (apoptosis).[J]. Immunology. 1985, 56(2): 351-358.
[96] Platt N, Suzuki H, Kurihara Y, et al. Role for the class A macrophage scavenger receptor in the phagocytosis of apoptotic thymocytes in vitro.[J]. Proc Natl Acad Sci U S A. 1996, 93(22): 12456-12460.
[97] Devitt A, Moffatt O D, Raykundalia C, et al. Human CD14 mediates recognition and phagocytosis of apoptotic cells.[J]. Nature. 1998,392(6675): 505-509.
[98] Luciani M F, Chimini G. The ATP binding cassette transporter ABC1, is required for the engulfment of corpses generated by apoptotic cell death.[J]. EMBO J. 1996,15(2): 226-235.
[99] Oka K, Sawamura T, Kikuta K, et al. Lectin-like oxidized low-density lipoprotein receptor 1 mediates phagocytosis of aged/apoptotic cells in endothelial cells.[J]. Proc Natl Acad Sci U S A. 1998, 95(16): 9535-9540.
[100] Ramprasad M P, Fischer W, Witztum J L, et al. The 94- to 97-kDa mouse macrophage membrane protein that recognizes oxidized low density lipoprotein and phosphatidylserine-rich liposomes is identical to macrosialin, the mouse homologue of human CD68.[J]. Proc Natl Acad Sci U S A. 1995,92(21): 9580-9584.
[101] Dransfield I, Buckle A M, Savill J S, et al. Neutrophil apoptosis is associated with a reduction in CD16 (Fc gamma RIII) expression.[J]. J Immunol. 1994, 153(3): 1254-1263.
[102] Hart S P, Ross J A, Ross K, et al. Molecular characterization of the surface of apoptotic neutrophils: implications for functional downregulation and recognition by phagocytes.[J]. Cell Death Differ. 2000, 7(5): 493-503.
[103] Hart S P, Smith J R, Dransfield I. Phagocytosis of opsonized apoptotic cells: roles for 'old-fashioned' receptors for antibody and complement.[J]. Clin Exp Immunol. 2004,135(2): 181-185.
[104] Hart S P, Alexander K M, Dransfield I. Immune complexes bind preferentially to Fc gamma RIIA (CD32) on apoptotic neutrophils, leading to augmented phagocytosis by macrophages and release of proinflammatory cytokines.[J]. J Immunol. 2004, 172(3): 1882-1887.
[105] Ren Y, Silverstein R L, Allen J, et al. CD36 gene transfer confers capacity for phagocytosis of cells undergoing apoptosis.[J]. J Exp Med. 1995,181(5): 1857-1862.
[106] Rossi A G, Mccutcheon J C, Roy N, et al. Regulation of macrophage phagocytosis of apoptotic cells by cAMP.[J]. J Immunol. 1998,160(7): 3562-3568.
[107] Liu Y, Cousin J M, Hughes J, et al. Glucocorticoids promote nonphlogistic phagocytosis of apoptotic leukocytes.[J]. J Immunol. 1999, 162(6): 3639-3646.
[108] Ogasawara J, Watanabe-Fukunaga R, Adachi M, et al. Lethal effect of the anti-Fas antibody in mice.[J]. Nature. 1993, 364(6440): 806-809.
[109] Hagimoto N, Kuwano K, Miyazaki H, et al. Induction of apoptosis and pulmonary fibrosis in mice in response to ligation of Fas antigen.[J]. Am J Respir Cell Mol Biol. 1997,17(3):272-278.
[110] Sawatzky D A, Willoughby D A, Colville-Nash P R, et al. The involvement of the apoptosis-modulating proteins ERK 1/2, Bcl-xL and Bax in the resolution of acute inflammation in vivo.[J]. Am J Pathol. 2006,168(1): 33-41.
[111] Rossi A G, Sawatzky D A, Walker A, et al. Cyclin-dependent kinase inhibitors enhance the resolution of inflammation by promoting inflammatory cell apoptosis.[J]. Nat Med. 2006, 12(9): 1056-1064.
[112] Rossi A G, Hallett J M, Sawatzky D A, et al. Modulation of granulocyte apoptosis can influence the resolution of inflammation.[J]. Biochem Soc Trans. 2007, 35(Pt 2): 288-291.
[113] Newman S L, Henson J E, Henson P M. Phagocytosis of senescent neutrophils by human monocyte-derived macrophages and rabbit inflammatory macrophages.[J]. J Exp Med. 1982, 156(2): 430-442.
[114] Jersmann H P, Ross K A, Vivers S, et al. Phagocytosis of apoptotic cells by human macrophages: analysis by multiparameter flow cytometry.[J]. Cytometry A. 2003, 51(1):7-15.
[115] Potter P K, Cortes-Hernandez J, Quartier P, et al. Lupus-prone mice have an abnormal response to thioglycolate and an impaired clearance of apoptotic cells.[J]. J Immunol. 2003,170(6): 3223-3232.
[1]Hart S P,Dransfield I,Rossi A G.Phagocytosis of apoptotic cells.[J].Methods.2008,44(3):280-285.
[2]Lauber K,Blumenthal S G,Waibel M,et al.Clearance of apoptotic cells:getting rid of the corpses.[J].Mol Cell.2004,14(3):277-287.
[3]Licht R,Jacobs C W,Tax W J,et al.An assay for the quantitative measurement of in vitro phagocytosis of early apoptotic thymocytes by murine resident peritoneal macrophages.[J].J Immunol Methods.1999,223(2):237-248.
[4]张卓然.培养细胞学与细胞培养技术:Cultural Cytology and Technique for Cell Culture[Z].上海:上海科学技术出版社,200412488.
[5]苑辉卿.医学细胞分子生物学实验[Z].北京:科学出版社,2007212.
[6]Shui Y B,Sasaki H,Pan J H,et al.Morphological observation on cell death and phagocytosis induced by ultraviolet irradiation in a cultured human lens epithelial cell line.[J].Exp Eye Res.2000,71(6):609-618.
[7]Castillo E C,Kouri J B.A new role for chondrocytes as non-professional phagocytes.An in vitro study.[J].Microsc Res Tech.2004,64(3):268-269.
[8]Peter A T,Dhanasekaran N.Apoptosis of granulosa cells:a review on the role of MAPK-signalling modules.[J].Reprod Domest Anim.2003,38(3):209-213.
[9]Palumbo A,Yeh J.In situ localization of apoptosis in the rat ovary during follicular atresia.[J].Biol Reprod.1994,51(5):888-895.
[10]胡野.细胞凋亡的分子医学[Z].北京:军事医学科学出版社,2002567.
[11]Messmer U K,Pfeilschifter J.New insights into the mechanism for clearance of apoptotic cells.[J].Bioessays.2000,22(10):878-881.
[12]Guzik K,Potempa J.Friendly fire against neutrophils:proteolytic enzymes confuse the recognition of apoptotic cells by macrophages.[J].Biochimie.2008,90(2):405-415.
[13]Zhang X J,Zheng G G,Ma X T,et al.Effects of various inducers on the expression of P2X7receptor in human peripheral blood mononuclear cells.[J].Sheng Li Xue Bao.2005,57(2):193-198.
[14]张容昶.中国的牦牛=China:The Yak:China:The Yak[Z].兰州:甘肃科学技术出版社,1998386.
[15]刘振魁.青海牦牛的繁殖特性[J].1981:5-7.
[16]牦牛繁殖育种编写组.牦牛繁殖力[J].1983(2):51-534.
[17]姜端生等.中甸牦牛繁殖现状调查[J].1985(3):25-28.
[18]牦牛繁殖育种编写组,牦牛繁殖与育种[J].中国牦牛.1982(3):73-82.
[19]Cui Y,Yu S J.An anatomical study of the internal genital organs of the yak at different ages.[J].Vet J.1999,157(2):192-196.
[20]Cui Y.Some physiological index in blood of yak at different ages(in Chinese).[J].1998,4.
[21]蒙学莲,崔燕,余四九,等.牦牛发情周期中卵巢卵泡发育状况的组织学观察[J].2006,36(1):57-61.
[22]Cui Y,Yong Y H,Yu S J.Follicular morphology in Yak in early pregnancy.Yak production in central asian highlands.Session Ⅳ:Reproduction and Physiology.[Z].2004.
[23]Yong Y H,Cui Y,Yu S J.Histological characteristics of corpus luteum in Yak during early pregnancy.YAK PRODUCTION IN CENTRAL ASIAN HIGHLANDS.Session IV:Reproduction and Physiology.[Z]. 2004.
[24] Yu S J, Chen B X. Endocrine changes just before the breeding season and during estrus in yak[J]. 1994:277-280.
[25] Yu S J, Huang Y M, Chen B X. Reproductive patterns of the yak. III. Levels of progesterone and oestradiol-17 beta during pregnancy and the periparturient period.[J]. Br Vet J. 1993, 149(6): 595-602.
[26] Yu S J, Huang Y M, Chen B X. Reproductive patterns of the yak. II. Progesterone and oestradiol-17 beta levels in plasma and milk just before the breeding season; also during normal and short oestrous cycles.[J]. Br Vet J. 1993, 149(6): 585-593.
[27] Yu S J, Huang Y M, Chen B X. Reproductive patterns of the yak. I. Reproductive phenomena of the female yak.[J]. Br Vet J. 1993, 149(6): 579-583.
[28] Yu S J, Ju X, Wang L, et al. Successful embryo transfer in Tianzhu white yak using standard protocol [J]. Science in China Series C: Life Sciences. 2007, 50(5): 655-659.
[29] Zhao H S, Yu S J, Zhao Q. Quantitative measurement of in vitro phagocytosis of apoptotic granulosa cells by monocytes in mice[J]. Sheng Li Xue Bao. 2009,61(2): 194-199.
[30] 中国牦牛学[Z]. 四川科学技术出版社,1989188-206.
[31] 青海省畜禽品种志[Z]. 青海人民出版社,198848-53.
[32] Dransfield I, Stocks S C, Haslett C. Regulation of cell adhesion molecule expression and function associated with neutrophil apoptosis.[J]. Blood. 1995, 85(11): 3264-3273.
[33] Savill J S, Wyllie A H, Henson J E, et al. Macrophage phagocytosis of aging neutrophils in inflammation. Programmed cell death in the neutrophil leads to its recognition by macrophages.[J]. J Clin Invest. 1989, 83(3): 865-875.
[34] Stern M, Savill J, Haslett C. Human monocyte-derived macrophage phagocytosis of senescent eosinophils undergoing apoptosis. Mediation by alpha v beta 3/CD36/thrombospondin recognition mechanism and lack of phlogistic response.[J]. Am J Pathol. 1996,149(3): 911-921.
[35] Fadok V A, Bratton D L, Konowal A, et al. Macrophages that have ingested apoptotic cells in vitro inhibit proinflammatory cytokine production through autocrine/paracrine mechanisms involving TGF-beta, PGE2, and PAF.[J]. J Clin Invest. 1998,101(4): 890-898.
[36] Savill J, Dransfield I, Gregory C, et al. A blast from the past: clearance of apoptotic cells regulates immune responses.[J]. Nat Rev Immunol. 2002,2(12): 965-975.
[37] Dransfield I, Buckle A M, Savill J S, et al. Neutrophil apoptosis is associated with a reduction in CD16 (Fc gamma RIII) expression.[J]. J Immunol. 1994,153(3): 1254-1263.
[38] Hart S P, Ross J A, Ross K, et al. Molecular characterization of the surface of apoptotic neutrophils: implications for functional downregulation and recognition by phagocytes.[J]. Cell Death Differ. 2000, 7(5): 493-503.
[39] Hart S P, Smith J R, Dransfield I. Phagocytosis of opsonized apoptotic cells: roles for 'old-fashioned' receptors for antibody and complement.[J]. Clin Exp Immunol. 2004,135(2):181-185.
[40] Hart S P, Alexander K M, Dransfield I. Immune complexes bind preferentially to Fc gamma RIIA (CD32) on apoptotic neutrophils, leading to augmented phagocytosis by macrophages and release of proinflammatory cytokines.[J]. J Immunol. 2004,172(3): 1882-1887.
[41] Ren Y, Silverstein R L, Allen J, et al. CD36 gene transfer confers capacity for phagocytosis of cells undergoing apoptosis.[J]. J Exp Med. 1995,181(5): 1857-1862.
[42] Rossi A G, Mccutcheon J C, Roy N, et al. Regulation of macrophage phagocytosis of apoptotic cells by cAMP.[J]. J Immunol. 1998, 160(7): 3562-3568.
[43] Liu Y, Cousin J M, Hughes J, et al. Glucocorticoids promote nonphlogistic phagocytosis of apoptotic leukocytes.[J]. J Immunol. 1999, 162(6): 3639-3646.
[44] Ogasawara J, Watanabe-Fukunaga R, Adachi M, et al. Lethal effect of the anti-Fas antibody in mice.[J]. Nature. 1993,364(6440): 806-809.
[45] Hagimoto N, Kuwano K, Miyazaki H, et al. Induction of apoptosis and pulmonary fibrosis in mice in response to ligation of Fas antigen.[J]. Am J Respir Cell Mol Biol. 1997, 17(3): 272-278.
[46] Sawatzky D A, Willoughby D A, Colville-Nash P R, et al. The involvement of the apoptosis-modulating proteins ERK 1/2, Bcl-xL and Bax in the resolution of acute inflammation in vivo.[J]. Am J Pathol. 2006,168(1): 33-41.
[47] Rossi A G, Sawatzky D A, Walker A, et al. Cyclin-dependent kinase inhibitors enhance the resolution of inflammation by promoting inflammatory cell apoptosis.[J]. Nat Med. 2006, 12(9): 1056-1064.
[48] Rossi A G, Hallett J M, Sawatzky D A, et al. Modulation of granulocyte apoptosis can influence the resolution of inflammation.[J]. Biochem Soc Trans. 2007,35(Pt 2): 288-291.
[49] Newman S L, Henson J E, Henson P M. Phagocytosis of senescent neutrophils by human monocyte-derived macrophages and rabbit inflammatory macrophages.[J]. J Exp Med. 1982,156(2): 430-442.
[50] Jersmann H P, Ross K A, Vivers S, et al. Phagocytosis of apoptotic cells by human macrophages: analysis by multiparameter flow cytometry.[J]. Cytometry A. 2003,51(1): 7-15.
[51] Potter P K, Cortes-Hernandez J, Quartier P, et al. Lupus-prone mice have an abnormal response to thioglycolate and an impaired clearance of apoptotic cells.[J]. J Immunol. 2003,170(6): 3223-3232.
