DY NAM IC CHANGE OF EP ID ERM AL STEM CELL S IN THE WOUND HEAL ING COURSE OF D IABETIC RATSg
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- Αίγλη Ρέντης
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1 693 (diabetes m ellitus, DM ) (ep iderm al stem cells, ESC s), ESC s DM 48 W istar,, g DM, 24 DM DM 115 cm 118 cm 2154 cm 2 ( 96 ), , ;, H E 19 (keratin 19, K19) Β1, , 24148% 3137% 50146% 1126% 92182% 2112% 99141% 0166%, DM 2143% 1102% 40159% 1165% 80177% 3157% 85140% 0194%, (P < 0101) , Λm,DM Λm, 3 21, (P < 0101) , K19 (po sitive unit, PU ) 91168% 93114% 72127% 70131%, DM 40129% 40179% 29194% 10137% ; Β1 PU 49160% 91116% 77113% 57117%,DM 38194% 24116% 61136% 38183%,, DM K19 Β1 PU, (P < 0105) ESC s DM : R R 332 : A DY NAM IC CHANGE OF EP ID ERM AL STEM CELL S IN THE WOUND HEAL ING COURSE OF D IABETIC RATSg T IA N X iaohong, B A I S hu ling, T IA N W ei, et al. Institu te of T issue E ng ineering, Colleg e of B asic M ed ical S ciences, Ch ina M ed ical U niversity, S heny ang L iaoning, , P. R. Ch ina. E 2m ail: tianx iaohong 2123@ hotm ail. com Corresp ond ing au thor: B A I S hu ling, E 2m ail: B ais hu ail. com Abstract Objective To observe the ep iderm al w idth, the amount variation and distribution character of ep iderm al stem cells (ESC s) and the w ound healing rate at different periods of diabetes m ellitus(dm ) rats after traum a, then to study the co rrelation of them. M ethods Fo rty2eigh tw istar rats w ere divided into DM group and no rm al contro l group random ly (n= 24). T he DM rats w ere induced by strep tozocin (ST Z) and then m ade ch ronic healing w ound by special perfo rex. A t the 3rd, 4th, 7th, 14th and 21st days after traum a, the healing rate w as calculated, the w ound edge and granulation tissue w ere obtained fo r haem atoxylin2eo sin (H E) staining and imm unoh istochem ical staining of keratin 19 (K19) and Β1 in tegrin. T hen the ep iderm al w idth, the area and the gradation value of po sitive un it ( PU ) w ere m easured. Results A t the 3rd, 7th, 14th and 21st days after traum a, the w ound healing rates of no rm al rats w ere 24148% 3137%, 50146% 1126%, 92182% 2112% and 99141% 0166% respectively, w h ile tho se of DM rats w ere 2143% 1102%, 40159% 1165%, 80177% 3157% and 85140% 0194% respectively, show ing significant differences (P < 0101). Befo re traum a, there w as no significant difference in the ep iderm al w idth betw een no rm al rats and DM rats (P > 0105). How ever, at the 3rd, 7th, 14th and 21st days after traum a, the ep iderm al w idth s of no rm al rats w ere , , and Λm respectively, w h ile tho se of DM rats w ere , , and Λm respectively, the ep iderm al w idth s in DM rats w ere significantly low er than tho se in no rm al rats (P < 0101). T he average PU value of K19 in no rm al rats w ere 91168%, 93114%, 72127% and 70131% respectively, w h ile tho se in DM rats w ere 40129%, 40179%, 29194% and 10137% respectively. T he average PU value of Β1 in tegrin in no rm al rats w ere 4916%, 91116%, 77113% and 57117% respectively, w h ile tho se in DM rats w ere 38194%, 24116%, 61136% and 38183%. T he resu lts indicated that the average PU values of K19 and Β1 integrin in DM rats w ere significantly low er than tho se in no rm al rats (P < 0. 05). Conclusion T he amount and activity decrease of ESC s m ay be one of the impo rtant m echanism s of difficult recovering w ounds of DM ratṡ Key words D iabetes m ellitus w ound W ound healing Ep iderm al stem cells D ynam ic change R at : (, ) :,,, :,E2m ail: BaiShuling@ho tm ail. com
2 694 Ch inese Journal of Reparative and Reconstructive Surgery, July 2007, V o l. 21, N o. 7 (ep iderm al stem cells, ESC s), Β1 19 (keratin 19, K19) ESC s [ 123 ] (diabetes m ellitu s, DM ),, [ 4 ], ESC s ESC s DM W istar 48,, g, (strep tozocin, ST Z, ), K19 Β1 SABC DAB (, ) W istar DM, DM [ 5 ] DM 12 h, ST Z (65 m ggkg), 72 h (, ),, mm o lgl,, DM (1 4 U g ) DM mm o lgl 3, 115 cm 118 cm 2154 cm 2 ( 96 ),,, % 65% ,, , 6,, 4%,,, H E K19 Β1, 48 [ 6 ] :, H 2O m in, 0101 m o lgl PBS 0101 m o lgl (ph 610), 20 m in (Β , K ), 4, PBS,, m in, PBS,, m in, DAB,, PBS, , M etam o rphgd P10gBX41, : = [ ( - ) g ] 100% [ 7 ] 200 (BX51, O lym pus, ),, M etam o rphgd P10gBX41 H E 3, Β1 K19,,, (m + n) (n) ( m + n) (GA ) Α (GΑ), (po sitive un it, PU ) : PU = 100 g GA 2GΑgg { Gm ax [ 1- ng(m + n) ]}, %, Gm ax, Gm ax= 256 [ 8 ] PU ESC s 1. 4 SPSS 1310, t, P < ,,,,, ;, ; 50% 7, DM 21, 99141% ( 1),DM,,,,,,,,,,, 50% 7 1 4, 2 1,
3 695 Tab. 1 (n= 6, x θ s) 1 The wound healing rate and the epidermal width at differen t per iods af ter trauma (n= 6, x θ s) ( ) T im e (d) Group W ound healing rate (% ) Ep iderm al w idth (Λm ) 21 N o rm al contro l group DM DM group P < Compared w ith no rm al contro l group, P < % ( 1) ,, ( 1), 7, DM 14, 21, DM, (P < 0. 01) 21, 3 4,,,,, ( 2 a) ; DM, 1 2,,,,,,,,, ( 2 b) F ig. 1 (Λm) 1 The dynam ic change of epidermal width (Λm) 2. 3 DM K19 Β1 ( 3 4), DM,, , K19 PU 91168% 93114% 72127% 70131%, DM 40129% 40179% 29194% 10137% ; Β1 PU 49160% 91116% 77113% 57117%, DM 38194% 24116% 61136% 38183%,,DM K19 Β1 PU, (P < 0105) K19 Β1 PU ST Z Β, DM : : DM, ; :,,, :,,, DM [ 9 ],, [ 10, 11 ] DM,, DM [ 12 ] :,, ; [ 13 ] :,, ; (m atrix m etallop ro teinase, MM P s) DM [ 14 ] :,MM P s,,mm P s, ;
4 696 Ch inese Journal of Reparative and Reconstructive Surgery, July 2007, V o l. 21, N o (HE 200) 1: 2: a b DM 3 14, K19 ( 200) a, b DM, 4 7 Β1 ( 200), a b DM F ig. 2 H istolog ical observation of skin s at the 21 st day af ter in jury (HE 200) 1: Cuticular layer 2: D erm ic layer a In no rm al contro l group b In DM group F ig. 3 The expression of K19 in each group 14 d af ter in jury ( immunohistochem ical sta in ing 200) a In no rm al contro l group, the arrow s indicated the po sitive stainning cells, w h ich lie in the bulge of hair fo llicle and sebaceous glands b In DM group, the arrow s indicated the po sitive stainning cells, w h ich lie in the basal layer of ep iderm is, bulge of hair fo llicle, and sebaceous glands F ig. 4 The expression of Β1 in tegr in in each group 7 d af ter in jury ( immunohistochem ical 200) T he arrow s indicated the po sitive stainning cells, w h ich lie in the basal layer of ep iderm is and sebaceous glands a In no rm al contro l group b In DM group 5 K19 PU F ig. 5 The dynam ic change of K19 s PU value 6 Β1 PU F ig. 6 The dynam ic change of Β1 in tegr in s PU value (ep iderm al grow th facto r recep to r, EGFR ) [ 15 ] : EGFR,,, ; [ 16 ] :,,DM, [ 17, 18 ], ESC s, DM,, DM?,,, DM,,, DM,,, 7, DM 14, DM
5 697,,, DM, (P < 0101) 50% 7, 7 14 K19 ESC s,dm K19 PU, DM, [ 19 ] ; DM K19 PU 14, 7, DM,, Β1, ESC s, ESC s, Β1, 6,,DM Β1 PU, ESC s,,,dm (P < 0101), DM ESC s [ 4 ],DM g, DM, DM ( ) ESC s, ESC s DM L I [ 20 ] ESC s, ESC s, ESC s, DM ESC s,, DM,,, ESC s, 4 1 Janes SM, Low ell S, H utter C. Ep iderm al stem cells. J Patho l, 2002, 197 (4) : Slack JM. Stem cells in ep ithelial tissues. Science, 2000, 287 (5457) : ,,.., 2004, 24 (2) : ,,,.., 2005, 9 (3) : ,.., 2005, 19 (2) : ,,,.., 2002, 16 (1) : N agelschm idt M, Becker D, Bonninghoff N, et al. Effect of fibronectin therapy and fibronectin deficiency on wound healing: a study in rats. J T raum a, 1987, 27 (11) : (g )., 1995, 4 (1) : ,,,.., 1997, 17 (4) : , 2000, 16 (6) : ,.., 2006, 12 (5) : ,,.., 2003, 7 (17) : ,,,. g., 2005, 21 (9) : W all SJ, Samp son M J, L evell N, et al. Elevated m atrix m etallop ro teinase22 and 23 p roduction from hum an diabetic derm al fibroblasts. B r J D erm ato l, 2003, 149 (1) : ,,.., 2003, 7 (6) : ,.., 2005, 25 (6) : Chbinou N, F renette J. Insulin2dependent diabetes impairs the inflamm ato ry response and delays angiogenesis fo llow ing A chilles tendon injury. Am J Physio l Regul Integr Comp Physio l, 2004, 286 (5) : A l2m ashat HA, Kandru S, L iu R, et al. D iabetes enhances mrna levels of p roapop to tic genes and caspase activity, w hich contribute to impaired healing. D iabetes, 2006, 55 (2) : ,,,.., 2004, 20 (8) : L I J ianfu, FU X iaobing, SH EN G Zhiyong, et al. Ectop ia of Ep iderm al Stem Cells on W ound Edge D uring W ound H ealing P rocess. Chinese Journal of Reparative and Reconstructive Surgery, 2006, 20 (3) : ( : : ) ( : )
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