011 mol/ L,pH410 - C(protein kinase C, PKC) EL Nahas ( Shefflield Kidney Institute, UK) ; ABC, ma ; Whatman 3mm. (diabetic nephropathy, DN), DN

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1 1238 hinese Journal of Pathophysiology 2003,19 (9) : [ ] (2003) , 1, 1, 1, 2 ( 1, 2, ) [ ] : (PK ) - 1 (TGF - 1 ) - (- SM), 3 : ( ),1 ( ),1 ( ) 2 ( ) Western PK,TGF SM,,, : PK TGF - 1 ( P < 0105), PK, TGF SM 3, : PK,,, TGF SM, [] ; - ; ; ; ; [] R [] (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 SM, N, 120 mg/ kg, 24 h, - 72 h, 1 Glucometer 4 (aer ), 111 Wistar 40, mmol/ L g, , Sigma, 1, mol/ L,pH410 -,24 h, 6 % ; PKTGF ,,, Santa ruz ; - SM - 80 Western, [ ] [ ] [ ] (No ) EL Nahas ( Shefflield Kidney Institute, UK) ;, ma ; Whatman 3mm ( ) ; 1 ( ) ; 1 ( ) ; 2 ( ) %2 4 %, ; ;HO - PP ; GPO

2 PO ;PK, TGF -, 3m,HE PS - SM PK TGF, 5, 4m,, - - ( ) PK TGF 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 SM ( ) - SM ( ) 1 PK, TGF SM Tab 1 The expression of PK, TGF SM in glomeru2 lus( gx s. n = 8) Group PK TGF SM P < vs group ; P < 0105 vs the group before ,,, 1 24 h (cr), ( ) PK TGF SM, 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 < vs group PK TGF SM Tab 3 The correlation between expression of PK,TGF - 1,- SM in glomeruli and impairment of kidney 1 TGF SM cr 24 h urine protein KW/ W PK TGF SM 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 < Western blot PK TGF - 1, 5 6 Fig 6 Western blot of TGF - 1 (two samples in each group). 6 TGF - 1 Western 5,,,

3 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) 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) : 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) : 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 , TGF - 1 [ 8 ] Hirofumi M. Phenotypic modulation of the mesangium reflected by contractile proteins in diabetes[j ]. iabetes,1996,45 (7) : Hirofumi [8 ] N 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) : [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 SM plex nephritis[j ]. J lin Invest,1991,87 (17) : , PKTGF - [11 ] Zhang GZ, Moorhead PJ, EI Nahas M. Myofibroblasts and - SM,the progression of experimental glomerulonephritis [J ]. Exp SM, Nephrol, 1995,3 (16) : [12 ],,,. - SM [10-12 ],,, N,, TGF SM, [ ] [1 ] Nishizuka Y. Intracellular singnaling by hydrosis of phospho2 lipid and activation of PK[J ]. Science,1992, 258 : [2 ] Nishizuka Y. The molecular heterogeneity of PK and its im2 plication for cellular regulation[j ]. Nature, 1988,334 : [3 ] Evaostlund M. Expression of PK isoforms in renal tissue[j ]. Kidney Int, 1995,47 (3) : sion of PK isoforms in streptozotocin - induced diabetic rats [J ]. Kidney Int, 1999,56 (5) : scriptional regulation in activated mesangial cells in vivo[j ]. [J ].,2001, 17 (6) :

4 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 , 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

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