1238 hinese Journal of Pathophysiology 2003,19 (9) :1238-1241 [ ] 1000-4718 (2003) 09-1238 - 04 3 1, 1, 1, 1, 2 ( 1, 2, 550004) [ ] : (PK ) - 1 (TGF - 1 ) - (- SM), 3 : ( ),1 ( ),1 ( ) 2 ( ) Western PK,TGF - 1 - SM,,, : PK TGF - 1 ( P < 0105), PK, TGF - 1 - SM 3, : PK,,, TGF - 1 - SM, [] ; - ; ; ; ; [] R692. 3 [] (protein kinase, PK), [1 ],10 PK [2 ], ; Western Sig2 PK, PK PK [3 ] PK, ;,, (diabetic nephropathy, N), PK N [4-6 ], 113 YY8 721 N () ; ( ), PK 2 - (- smooth muscle actin, 211 4, 8 - SM) N PK,TGF - 1 - SM, N, 120 mg/ kg, 24 h, - 72 h, 1 Glucometer 4 (aer ), 111 Wistar 40, 152-287 1617 mmol/ L g, 213 112, Sigma, 1,1 2 011 mol/ L,pH410 -,24 h, 6 % ; PKTGF - 1-20,,, Santa ruz ; - SM - 80 Western, [ ] 2002-06 - 27 [ ] 2002-10 - 09 3 [ ] (No. 2000-8) EL Nahas ( Shefflield Kidney Institute, UK) ;, ma ; Whatman 3mm ( ) ; 1 ( ) ; 1 ( ) ; 2 ( ) 212 6 %2 4 %, ; ;HO - PP ; GPO
1239 - PO ;PK, TGF -, 3m,HE PS - SM PK TGF, 5, 4m,, - - ( ) PK TGF - 1 - SM, PS, (200 ) 15, PK TGF - 1 Western 2 30 mg 8 %SS - PGE,,5 %, PS, 8 h, PS,, 3 gx s,studentπs t, 1 PK, TGF - 1 - SM - 1 2 ( ) - SM 1 1 2 3 ( ) 1 PK, TGF - 1 - SM Tab 1 The expression of PK, TGF - 1 - SM in glomeru2 lus( gx s. n = 8) Group PK TGF - 1 - SM 1. 383 0. 698 0 0 4. 808 2. 109 3 1. 313 0. 124 3 0 7. 308 1. 750 3 4. 582 0. 826 3 0 14. 183 3. 513 3 8. 216 1. 642 3 6. 625 1. 242 3 P < 0. 05 vs group ; P < 0105 vs the group before. 2 24 2,,, 1 24 h (cr), 2 3 2 ( ) PK TGF - 1 - SM, 3 2 24 h Tab 2 Level of blood glucose,cholesterol,triglycerides,cr,24 h urine protein in various group ( gx s, n = 8) Group lood glucose (mmol/ L) holesterol (mmol/ L) Triglycerides (mmol/ L) cr (ml/ min) 24 h urine protein (mg) 3 P < 0. 05 vs group. 4. 268 1. 154 0. 683 0. 335 0. 620 0. 200 0. 430 0. 136 11. 450 3. 800 24. 487 6. 035 1. 048 0. 130 3 0. 873 0. 228 1. 788 0. 609 3 46. 905 16. 658 3 28. 947 5. 112 1. 313 0. 276 3 1. 370 0. 876 3 2. 946 0. 383 3 81. 162 23. 533 3 27. 406 4. 518 1. 625 0. 239 3 1. 588 0. 326 3 4. 724 1. 156 3 106. 387 13. 802 3 3 PK TGF - 1 - SM Tab 3 The correlation between expression of PK,TGF - 1,- SM in glomeruli and impairment of kidney 1 TGF - 1 - SM cr 24 h urine protein KW/ W PK 0. 484 3 0. 768 3 0. 978 33 0. 937 33 0. 969 33 TGF - 1 0. 368 3 0. 995 33 - SM 0. 914 33 Fig 5 Western blot of PK(two samples in each group). 5 PKWestern Numbers in this table were correlation coefficients. 3 P < 0. 05, 33 P < 0101. 4 Western blot PK TGF - 1, 5 6 Fig 6 Western blot of TGF - 1 (two samples in each group). 6 TGF - 1 Western 5,,,
1240,, PK,,,,,, 4 4 ( ) 4 / Tab 4 The changes of body weight, kidney weight, KW/ W and Group diameter of glomerulus( gx s. n = 8) ody weight (g) Kidney weight (g) KW/ W iameter of glomerulus (m) 243121 11123 1103 0122 01004 01001 73. 00 2170 225114 13145 1114 0118 01005 01001 116. 00 3142 3 236136 16178 1125 0113 3 01006 01001 3 121162 3155 3 248175 18191 1144 0121 3 01008 01002 3 131175 4113 3 3 P < 0105 vs group. [4 ] Lee TS, Mochly - Rosen. ctivation of PK by elevation of glucose concentration ; phosal for a mechanism in the develop2, 1, ment of vascular complications[j ]. Proc Natl cad Sci US,,,Western 1989,86 (25) :5141-5150. PK, [5 ] Kang N, lexander G, Maasch, et al. ifferential expres2, PK 1 [6 ] Williams, Howard RL. Glucose - induced PK activity reg2 TGF - 1,Western TGF - 1 ulates arachidonic acid release and eicosanoid production by cultured glomerular mesangial cells[j ]. J lin Invest, 1993,,PK / Yamamoto [7 ] 92 (14) :2889-2895. N TGF - 1 [7 ] Yamamoto T, Noble N, ohen H, et al. The expression of,pk, TGF - in diabetic kidney[j ]. Kidney Int, 1996,49 (2) :461, N PK - 472., TGF - 1 [ 8 ] Hirofumi M. Phenotypic modulation of the mesangium reflected by contractile proteins in diabetes[j ]. iabetes,1996,45 (7) : Hirofumi [8 ] N 24 488-499. - SM,2 [9 ] Kawada N. Role of intron 1 in smooth muscle- actin tran2, - SM PK TGF - 1 Kawada [9 ] TGF - 1 Kidney Int, 1999,55 (6) :2338-2348. [10 ] Johnson RJ, Ida H, lpers E, et al. Expression of smooth - SM, muscle cell phenotype by rat mesengial cells in immune com2,pk TGF - 1 - SM plex nephritis[j ]. J lin Invest,1991,87 (17) :847-858., PKTGF - [11 ] Zhang GZ, Moorhead PJ, EI Nahas M. Myofibroblasts and - SM,the progression of experimental glomerulonephritis [J ]. Exp SM, Nephrol, 1995,3 (16) :308-318. [12 ],,,. - SM [10-12 ],,, N,, TGF - 1 - SM, [ ] [1 ] Nishizuka Y. Intracellular singnaling by hydrosis of phospho2 lipid and activation of PK[J ]. Science,1992, 258 : 607-614. [2 ] Nishizuka Y. The molecular heterogeneity of PK and its im2 plication for cellular regulation[j ]. Nature, 1988,334 : 661-665. [3 ] Evaostlund M. Expression of PK isoforms in renal tissue[j ]. Kidney Int, 1995,47 (3) :766-773. sion of PK isoforms in streptozotocin - induced diabetic rats [J ]. Kidney Int, 1999,56 (5) :1737-1745. scriptional regulation in activated mesangial cells in vivo[j ]. [J ].,2001, 17 (6) :569-570.
1241 Study on relationship between expression of PKin glomeruli and development of nephropathy in diabetic rats J I Zhong - hui 1, ZHNG Guo - zhong 1, GUO ing 1, GUI Hua - zhen 1, WN hang - wu 2 ( 1 epartment of Pathophysiology, 2 epartment of Pathology, Guiyang Medical ollege, Guiyang 550004, hina) [ STRT] IM : To observe the dynamic changes of expression of PK, TGF - 1 and- SM in glomeruli of diabetic rats induced by the alloxon and to invesitigate their roles in the diabetic nephropathy(n). METH2 OS : Rats were randomly divided into four groups : normal control group (group ), diabetic group of one week (group ), diabetic group of one month (group ), diabetic group of two months (group ). Immunohistochemistry and West2 ern blotting were used to detect the expression of PK, TGF - 1 and- SM in renal tissue of all groups. lood glu2 cose, triglycerides, cholesterol, creatinine and urine protein were analysed by chemical methods. The morphological changes of renal tissue were checked through microscopy. RESULTS : The expression of PKand TGF - 1 in renal tis2 sue of diabetic groups were increased comparing with those of nomal control group ( P < 0. 05). The mesangial cells ex2 pressed- SM in two months group. hronologically the expression of PK, TGF - 1 and- SM were positively correlative with each other and the impairment of kidney was also observed. ONL USIONS : uring the N process the expression of PKincreased. PKraised GFR and the permeability of glomerular filtration membrane which en2 hanced urinary albumin excretion. PKalso increased expression of TGF -and therefore to induce the expression of - SM. The appearance of- SM was a marker of the phenotypic transform of renal cells. [ KEY WORS] Protein kinase ; Transforming growth factor beta ; Smooth muscle actin ; Glomerulus ; iabetes mellitus ; Rats