693 (diabetes m ellitus, DM ) (ep iderm al stem cells, ESC s), ESC s DM 48 W istar,, 180 200 g DM, 24 DM DM 115 cm 118 cm 2154 cm 2 ( 96 ), 3 7 14 21, ;, H E 19 (keratin 19, K19) Β1, 3 7 14 21, 24148% 3137% 50146% 1126% 92182% 2112% 99141% 0166%, DM 2143% 1102% 40159% 1165% 80177% 3157% 85140% 0194%, (P < 0101) 3 7 14 21, 26143 3121 33129 3152 31153 3135 26101 3119 Λm,DM 23158 2133 31102 3138 33172 5149 21180 4102 Λm, 3 21, (P < 0101) 3 7 14 21, 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 58712 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, 110001, 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 ling @hotm 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 26143 3121, 33129 3152, 31153 3135 and 26101 3119 Λm respectively, w h ile tho se of DM rats w ere 23158 2133, 31102 3138, 33172 5149 and 21180 4102 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 : (, 110001) :,,, :,E2m ail: BaiShuling@ho tm ail. com
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 1 1. 1 7 W istar 48,, 180 200 g, (strep tozocin, ST Z, ), K19 Β1 SABC DAB (, ) 1. 2 1. 2. 1 48 W istar DM, 24 1. 2. 2 DM [ 5 ] DM 12 h, ST Z (65 m ggkg), 72 h (, ),, 3 16. 8 mm o lgl,, DM (1 4 U g ) DM 1110 1615 mm o lgl 3, 115 cm 118 cm 2154 cm 2 ( 96 ),,, 18 25 45% 65% 1. 3 1. 3. 1 3 7 14 21,, 1. 3. 2 3 7 14 21, 6,, 4%,,, H E K19 Β1, 48 [ 6 ] :, H 2O 2 5 10 m in, 0101 m o lgl PBS 0101 m o lgl (ph 610), 20 m in (Β1 1 100, K19 1 150), 4, PBS,, 37 20 m in, PBS,, 37 20 m in, DAB,, PBS, 1. 3. 3, 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 < 0105 2 2. 1,,,,, ;, ; 50% 7, DM 21, 99141% ( 1),DM,,,,,,,,,,, 50% 7 1 4, 2 1,
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 3 7 14 W ound healing rate (% ) Ep iderm al w idth (Λm ) 21 N o rm al contro l group DM DM group 24. 48 3. 37 26. 43 3. 21 50. 46 1. 26 33. 29 3. 52 92. 82 2. 12 31. 53 3. 35 99. 41 0. 66 26. 01 3. 19 2. 43 1. 02 3 23. 58 2. 33 3 40. 59 1. 65 3 31. 02 3. 38 80. 77 3. 57 3 33. 72 5. 49 85. 40 0. 94 3 21. 80 4. 02 3 3 P < 0. 01 3 Compared w ith no rm al contro l group, P < 0101 85140% ( 1) 2. 2 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,, 3 7 14 21, 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 5 6 3 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, ;
696 Ch inese Journal of Reparative and Reconstructive Surgery, July 2007, V o l. 21, N o. 7 2 21 (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
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