( 2smooth muscle actin, 2SM actin)

Transcription

1 UV2C 234, , 51 (2), Acta Physiologica Sinica 3 (, ) (220 W, 220 V, 50 Hz) UV2C (254 nm), 10 cm (smooth muscle cells, SM2 Cs),, ; / ; DNA UV2C SMCs SMCs : ; ; ; : Q2233 (smooth muscle cells, SMCs) (atherosclerosis, AS), SMCs AS,,, SMCs SMCs 2C(ultraviolet C, UV2C) SMCs,, SMCs [1 SMCs ] g SD,,, SMCs, SP 2 ( 2smooth muscle actin, 2SM actin) UV2C SMCs 4 7 SMCs,, 10 5 / ml 60 mm 48 h, ml D2Hank s, UV2C 10 cm,, 10 min, :,

2 2 : UV2C % DMEM, 5 % CO 2, UV2C SMCs 4 7 SMCs 48 h, UV2C, h, HE, UV2C SMCs, UV2 C h 4, D2Hank s 2 3,, 10 min 5, 10, ( n ), Quantimet 520 +, (cellular area, CA) (nuclear area, NA) (ratio of area, RA) 3 : RA = NA CA 100 %. ( x s), t, SPLM ( ) (Control) ; (UV2C). DNA :, UV2C SMCs, h, r/ min 5 min,, 4 ml PBS, 50 3 h,, 500 l,,, r/ min 5 min,, 2 (24 1) 500 l,,, r/ min 5 min,, 3 mol/ L 50 l 1 ml, - 20, 1000 r/ min 10 min,,, 100 l TE, RNase 5 l, min, 115 %, DNA : 11 SMCs SMCs,, SP 2SM actin, SMCs 21 UV2C SMCs 211 UV2C, 3 h, 12 h, 24 h HE, :,,,, 48 h 24 h, ; 72 h ( 1) 212 UV2C h SMCs (CA) (NA) / (RA) 1, UV2C h CA NA RA ( P < 0101), h CA NA RA ( P < 0101) UV2C SMCs

3 A1 SMCs ; B. UV2C 24 h SMCs ; C. UV2C 72 h SMCs Fig11 A. Normal SMCs (HE staining ; scale bar : 25 m). B1 Cell shinkage and condensation of cytoplasm and chromatin of SMCs at 24 h after exposure to UV2C (HE staining ; scale bar : 50 m). C. Appearance of apoptotic body in SMCs at 72 h after exposure to UV2C (HE staining ; scale bar : 25 m). 1 UV2C SMCs CA NA RA Table 1 Changes in CA, NA and RA after exposure to UV2C CA / m 2 Time after exposure to UV2C 24 h 48 h NA / m 2 Time after exposure to UV2C 24 h 48 h RA / % Time after exposure to UV2C 24 h 48 h Control UV2C % % % % P < 0101 as compared with control. CA : cellular area ; NA : nuclear area ; RA : ratio of area.,,, / ( ) %(control) ( ) %(24 h) ( ) %(48 h) 213 UV2C h DNA, 2 UV2C h DNA bp, (ladder), DNA, UV2C SMCs DNA 3 : UV2A ( nm) ; UV2B ( nm) ; UV2C ( 280 nm), DNA nm, 260 nm,

4 2 : UV2C SMCs DNA Fig12 DNA ladder of SMCs h after exposure to UV2C ; 21 Con2 trol ; after exposure to UV2C ; 41 Molecular mark. UV2C, UV2C, 254 nm, UV2C, UV2C, [2 UV2A UV2B ] DNA ( cyclobutane2type pyrimidine dimmer, CPD) 6,42 DNA DNA2 Sin2 glet, UV2C DNA, CPD,, Langerhans [3 ] [4 ] [5 HL260 U937 ], DNA Servomaa [6 ],, UV (L1Rn), ; Wright [5 ], UV HL260 U937 T Molt23 Molt24, UV DNA ; Gudar [7 ] UV 4 h, 20 h DNA Bazar [8 ], RNA UV T Jurkat, UV,, UV, Clusterin [9 ] fos/ jun [10 ] bcl22 [10,11 ] DNA P 53 [11 ], UV2C SMCs 10 min,,,

5 /,, Orlov [12 ] SMCs, SMCs, ; SMCs,, SMCs /, UV2C SMCs, UV2C,,, UV2C SMCs DNA [13, ], UV2C DNA,, UV2C SMCs DNA UV2C,, SMCs, SMCs [1 ] Ross R. The smooth muscle cell. II Growth of smooth muscle in culture and formation of elastic fibers. J Cell Biol, 1971, 50 : [2 ] Stapleton AE. Ultraviolet radiation and plants : burning questions. Plant Cell, 1992, 4 : [ 3 ] Hollis DE. Scheibner A. Ultrastructural changes in epidermal langerhans cells and melanocytes in response to ul2 traviolet irradiation in Australians of Aboriginal and Celtic desent. Br J Dermatol, 1988, 119 (1) : [4 ] Kane KS, Maytin EV. Ultraviolet B induced apoptosis of keratinocytes in murine skin is reduced by mild local hyperthermia. J Invest Dermatol, 1995, 104 (1) : [5 ] Wright SC, Wei QS, Zhong J, et al. Purification of a 242KD protase from apoptotic tumor cells that activates DNA fragmentation. J Exp Med, 1994, 180 (6) : [6 ] Servomaa K, Rytomaa T. UV light and ionizing radiations cause programmed death of rat chloroleukaemia cells by inducing retropositions of a mobile DNA element (L1Rn). Int J Radiat Biol, 1990, 57 (2) : [7 ] Gudar DE, Lucas AD. Spectral dependence of UV2induced immediate and delayed apoptosis : the role of mem2 brane and DNA damage. Photochem Photobiol, 1995, 62 (1) : [8 ] Bazar LS, Deng HJ. Ultraviolet B induced DNA fragmentation (apoptosis) in actived T2lymphocytes and Jurkat cells is augmented by inhibition of RNA and protein synthesis. Exp Hematol, 1992, 20 (1) : [9 ] French LE, Wohlwend A, Sappino AP, et al. Human clusterin gene expression is confined to surviving cells during in vitro programmed cell death. J Clin Invest, 1994, 93 (2) : [10 ] Gillardon F, Eschenfelder C, Uhlmann E, et al. Differential regulation of c2fos, fosb, c2jun, junb, bcl22 and bax expression in rat skin following single or chronic ultraviolet irradiation and in vivo modulation by antisens oligodeoxynucleotide superfusion. Oncogene, 1994, 9 (11) : [ 11 ] Jiang MC, Yang Yen HF, Lin J K, et al. Differential regulation of P 53, c2myc, Bcl22 and Bax protein expression during apoptosis induced by widely divergent stimuli in human hepatoblastoma cells. Oncogene, 1996, 13 (3) : [12 ] Orlov SN, Dam TV, Tremblay J, et al. Apoptosis in vascular smooth muscle cells : Role of cell shrinkage. Biochem Biophys Res Commun, 1996, 221 (3) : [13 ] Majno G, Joris I. Apoptosis, oncosis and necrosis : an overview of cell death. Am J Pathol, 1995, 146 (1) : 3 15.

6 Acta Physiologica Sinica Apr. 1999, 51 (2), ESTABL ISHMENT OF AN APOPTOSIS MODEL OF VASCULAR SMOOTH MUSCL E CELLS IND UCED BY ULTRAVIOLET2C RADIATION IN VITRO LI XIAO2DAN, LI J IN 3 ( Department of Internal Medicine of Guangzhou Medical College, PLA, Guangzhou ) ABSTRACT After a 102min exposure to 254 nm UV2C radiation, aortic SMCs of rat were found to undergo apoptosis, characterized by cell shrinkage, condensation of cytoplasm, mem2 brane blebbing, appearance of apoptotic body and DNA ladder in the agarose gel. There2 fore, UV2C radiation can be used to generate a useful model for the investigation of the molecular regulatory mechanisms underlying SMCs apoptosis. Key words : smooth muscle cells ; apoptosis ; ultraviolet ; atherosclerosis 3 Present address : Department of Histology and Embryology of the First Military Medical University, Guangzhou