http://www.kidney-international.
org
& 2007 International Society of Nephrology
The role of protein kinase C activation in diabetic
nephropathy
H Noh1 and GL King1
1
Research Division, Joslin Diabetes Center, Harvard Medical School, Boston, Massachusetts, USA
Diabetic nephropathy is the leading cause of end-stage renal
disease worldwide and an independent risk factor for all-
cause and cardiovascular mortalities in diabetic patients. New
insights into the molecular mechanisms that underlie the
development and progression of microvascular
complications of diabetes including nephropathy are
emerging rapidly from experimental and clinical studies.
Chronic hyperglycemia is a major initiator of diabetic
microvascular complications. Activation of diacylglycerol
(DAG)-protein kinase C (PKC) pathway, enhanced polyol
pathway, increased oxidative stress, and overproduction of
advanced glycation end products have all been proposed as
potential cellular mechanisms by which hyperglycemia
induces diabetic vascular complications. The DAG-PKC
pathway contributes to vascular function in many ways such
as the regulation of endothelial permeability,
vasoconstriction, extracellular matrix synthesis/turnover, cell
growth, angiogenesis, cytokine activation, and leukocyte
adhesion. We will briefly review the current knowledge base
regarding the pathogenic role for the activation of DAG-PKC
pathway in diabetic nephropathy and other microvascular
complications of diabetes. The results from animal studies
and key clinical studies investigating specific effects of the
PKC isoforms on the renal and other vascular tissues to
induce diabetic complications are also reviewed.
Kidney International (2007) 72, S49S53; doi:10.1038/sj.ki.5002386
KEYWORDS: diabetic nephropathy; diabetic microvascular complications;
protein kinase C
Correspondence: GL King, Room M4504, Joslin Diabetes Center, 1 Joslin
Place, Boston, Massachusetts 02215, USA.
E-mail: george.king@joslin.harvard.edu
Kidney International (2007) 72, S49S53
Diabetic nephropathy is the leading cause of end-stage renal
disease (ESRD) worldwide1 and an independent risk factor
for all-cause and cardiovascular mortalities in diabetic
patients.2 In patients with type I diabetes, 2540% will
develop diabetic nephropathy.3 Over 5% of newly diagnosed
patients with type II diabetes will already have diabetic
nephropathy and a further 3040% will develop diabetic
nephropathy4 with a high likelihood of progression to
ESRD. With the continuous expansion of the population
with type II diabetes, it is necessary to define the
pathophysiologic mechanisms for this disorder in order to
design specific therapies for prevention and reversal of
diabetic nephropathy.
Hyperglycemia has been shown to be the major risk factor
responsible for the development and progression of micro-
vascular complications of diabetes. The Diabetes Control and
Complications Trial in type I diabetes5 and the United
Kingdom Prospective Diabetes Study in type II diabetes6
demonstrated that intensive blood glucose control success-
fully delayed the onset and retarded the progression of
diabetic microvascular complications such as nephropathy,
retinopathy, neuropathy, and cardiovascular pathologies.
These data strongly suggested that hyperglycemia-induced
metabolic factors are responsible for diabetic vascular
complications. Multiple biochemical pathways have been
proposed to explain the adverse effects of hyperglycemia.
Activation of diacylglycerol (DAG)-protein kinase C (PKC)
pathway,7 enhanced polyol pathway,8 increased oxidative
stress,9 and overproduction of advanced glycation end
products10 have all been proposed as potential cellular
mechanisms by which hyperglycemia induces diabetic
vascular complications.
We will briefly review the current knowledge base
regarding the pathogenic role for the activation of DAG-
PKC pathway in diabetic nephropathy and other micro-
vascular complications of diabetes. Others and we have
shown that the activation of DAG-PKC pathway has been
associated with many vascular abnormalities in renal, retinal,
and cardiovascular tissues in diabetic animals and patients.
Changes in DAG-PKC activities are known to regulate
vascular functions in many ways such as regulation of
vascular permeability, contractility, extracellular matrix
(ECM) synthesis/turnover, cell growth and apoptosis,
angiogenesis, cytokine activation, and adhesions of mono-
S49

cytes and leukocytes, all of which are reported to be
abnormal in diabetes.11,12
PKC
PKC is a family of serine/threonine kinases that consist of 12
isoforms. PKC isoforms are classified according to whether
they contain domains that bind Ca2 or DAG, both of which
positively regulate the kinase activity. Conventional PKC (a,
b1, b2, and g) binds both Ca2 and DAG, novel PKC (d, e, Z,
y, m) binds DAG, but not Ca2 , and atypical PKC (z, l)
binds neither. The activation of conventional and novel PKC
isoforms require the correct phosphorylation of the isoforms
and the presence of cofactors such as Ca2 and DAG.
When properly phosphorylated, increases in Ca2 or
DAG rapidly or chronically will induce its translocation to
the membranous compartments of the cells to elicit
biological actions. Rapid and short-term increases of DAG
and Ca2 levels are usually induced by cytokines via the
activation of phospholipase C. Chronic activation of PKCs
require sustained elevations of DAG, which involves the
activation of phospholipase D/C or the de novo synthesis of
DAG.13 In hyperglycemic and diabetic state, all of these
pathways probably contribute to the activation of DAG-PKC
cascade.
MECHANISMS OF HYPERGLYCEMIA-INDUCED
PKC ACTIVATION
Increased total DAG contents have been reported in a variety
of tissues associated with diabetic vascular complications,
including renal glomeruli,1416 heart, aorta,17 and retina18
from diabetic animal models and patients. In addition, non-
vascular tissues such as liver,19 skeletal muscles,20 and
circulating monocytes have also displayed an increased
activation of DAG-PKC pathway in diabetic and insulin-
resistant animals. In many cell culture studies, using renal
mesangial cells,21,22 aortic17 and retinal18 endothelial cells, or
smooth muscle cells,17 increasing glucose levels in the media
from 5.6 to 22 mM elevated cellular DAG contents and
increased PKC activities. Interestingly, the increases of DAG
levels in cultured vascular cells usually require the exposure
to high glucose levels for hours to days suggesting that
induction of protein synthesis may be required.
Radioactive labeling studies have established that in-
creased DAG contents induced by diabetes or elevated
glucose levels occur through several pathways. One is by
increasing de novo DAG synthesis from the glycolytic
intermediate dihydroxyacetone phosphate, through reduc-
tion of the latter to glycerol-3-phosphate and stepwise
acylation.17 High glucose concentrations can cause increased
de novo DAG synthesis through several different metabolic
pathways. One proposed mechanism is that increased
synthesis of DAG is caused by inhibition of the glyco-
lytic enzyme glyceraldehyde-3-phosphate dehydrogenase,
thereby diverting upstream metabolites from glycolysis into
pathways of glucose over utilization. This results in increased
flux of dihydroxyacetone phosphate to DAG.9 There is also
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H Noh and GL King: Role of PKC activation in diabetic nephropathy
evidence that the increased levels of DAG can also be
derived from the actions of phospholipase D from phospha-
tidylcholine. Another supportive evidence that the increased
DAG levels are derived from pathways other than the
activation of phospholipase C is the findings that palmatate
and oleate are the major fatty acids in the elevated levels of
DAG. Furthermore, there is evidence that oxidants and
glycated products can also increase DAG levels and activate
PKC.
PKC ACTIVATION IN DIABETIC NEPHROPATHY
Diabetic nephropathy is characterized by initial glomerular
hyperfiltration, progressive accumulation of ECM in glomer-
ular mesangium and tubulointerstitium, and progressive
renal insufficiency. Hyperglycemia-induced metabolic, he-
modynamic, and possibly inflammatory factors are thought
to be mediating these injuries.23
Glomerular hyperfiltration
Increased glomerular filtration rate is described in the kidney
of diabetic patients and animal models.2426 This alteration is
likely to be the result of hyperglycemia-induced decreases in
arteriolar resistance,27,28 leading to an elevation of glomer-
ular filtration pressure. Multiple mechanisms have been
proposed to explain the increases in glomerular filtration rate
and filtration pressure, including an enhanced activity of
angiotensin II and prostaglandin productions.2931 Activation
of DAG-PKC pathway may play a role in both the
enhancement of angiotensin II actions29 and increases in
vasodilatory prostaglandins.30,31 The enhanced production of
prostaglandin E2 induced by diabetes and hyperglycemia
could be the result of sequential activation of PKC and
cytosolic phospholipase A2, a key regulator of arachidonic
acid synthesis.3235
Increases in the activities of nitric oxide (NO) may also
enhance glomerular filtration rate.36 Urinary excretion of
NO2 and NO3, metabolites of NO, has been reported to be
increased in diabetes of short duration,3638 possibly due to
enhanced expression of inducible NO synthase gene and
increased production of NO in mesangial cells.39 In addition,
both increases in inducible NO synthase gene expression and
NO production can be mimicked by PKC agonists and
inhibited by PKC inhibitors when induced by hyperglyce-
mia,39 suggesting that NO production might be increased in
diabetes through PKC-induced upregulation of inducible NO
synthase. However, it has been reported that NO and its
second messenger, cyclic guanosine monophosphate produc-
tions were decreased in diabetic rat glomeruli and PKC
inhibitors restore the glomerular cyclic guanosine monopho-
sphate production.40 Thus, it is possible that high glucose-
induced PKC activation may regulate renal hemodynamics by
increasing or decreasing NO production depending on
the type of cells and duration of hyperglycemia. Lastly,
recent studies have also shown that the expression of vascular
endothelial growth factor (VEGF) is also increased in the
glomeruli similar to the retina.41,42 PKC activation is
Kidney International (2007) 72, S49S53
H Noh and GL King: Role of PKC activation in diabetic nephropathy
also known to mediate several biological actions of
VEGF including its effect to increase capillary permeability.43
As VEGF can also increase blood flow and capillary
permeability, it is possible that increased VEGF levels and
activity may contribute to the abnormality of renal
hemodynamics.
Accumulation of ECM
Thickening of glomerular basement membrane and accumu-
lation of ECM in glomerular mesangium and tubulointer-
stitium are the hallmark of diabetic nephropathy. It has been
reported that high glucose increased production of type IV
collagen and fibronectin in mesangial cells. This finding could
be mimicked by phorbol ester, PKC agonist, and reversed by
general PKC inhibitors.22,44,45 Further, there are great deal of
support that hyperglycemia induced increases in oxidant
production via several mechanisms. In the kidney, several
reports have suggested that hyperglycemia can increase PKC
activities leading to activation of several isoforms of
nicotinamide adenine dinucleotide phosphate (reduced form;
NADPH) oxidases to produce the excessive oxidants. The
elevated levels of oxidants in combination with PKC induced
activations of mitogen-activated protein kinase will lead to the
overexpression of fibrotic growth factors (Figure 1). Many
studies have suggested that transforming growth factor
(TGF)-b1 play a key role in the accumulation of ECM.4649
It has been reported that PKC activation can increase the
production of ECM and TGF-b1 expression, and that PKC
inhibitors can prevent hyperglycemia or diabetes-induced
increases in ECM accumulation and TGF-b1 production in
mesangial cells or renal glomeruli.50 Recently, we have
reported that the excessive productions of TGF-b1 and
connective tissue growth factor, and ECM proteins induced
by diabetes were significantly reduced in PKCb-null mice as
compared with their wild-type controls.51 In addition,
Progression of diabetic nephropathy
Hyperglycemia (AGEs, oxidants, DAG synthesis)
PKC-
,+
MAPK
oxidases
CTGF/TGF-
Proteinuria, mesangium expansion, nephromegaly
Glomerular sclerosis
Diabetic nephropathy
Figure 1 | This Schema shows our hypothesis of diabetic
nephropathy. Hyperglycemia activates PKC and increase
connective tissue growth factor and TGF-b, which lead to mesangium
expansion and nephromegaly. Mesangium expansion due to
matrix deposition results in glomerular sclerosis and diabetic
nephropathy.
Kidney International (2007) 72, S49S53
diabetes also induced significantly less activation of NADPH
oxidase subunits, NOX 2 and 4 expression further support a
regulatory role of PKC activation in the excessive production
of oxidants, fibrotic factors, and ECMs observed in the renal
glomeruli in diabetic state.51
Vascular permeability and albuminuria
Increased vascular permeability is another characteristic
systemic vascular abnormality in diabetic animals, suggesting
endothelial cell dysfunction. PKC activation can directly
increase the permeability of albumin and other macromole-
cules through barriers formed by endothelial cells,52,53
probably by phosphorylating cytoskeletal proteins forming
the intracellular junctions.54,55 As well, PKC activation could
also regulate vascular permeability and neovascularization via
the expression of growth factors such as VEGF, which has
been demonstrated as a key factor in the pathogenesis of
diabetic retinopathy.56 It is also not likely that the elevation of
VEGF is responsible for the increases in capillary perme-
ability observed in diabetes since several tissues such as the
heart and peripheral limb vessels also have increases in
vascular permeability, yet VEGF expressions are decreased in
those tissues.42 In the kidney, however, the role of PKC
activation in the genesis of albuminuria is not clearly defined.
Given that the glomerular filtration barrier is a unique
structure composed of three different parts including the
glomerular endothelial cells, the basement membrane, and
the podocytes, consideration should be given to the alteration
of all of these components. Interestingly, little is known about
the effects of PKC activation on the biology of glomerular
endothelial cells and podocytes. Recently, it was shown that
hyperglycemia-induced downregulation of the negatively
charged basement membrane heparan sulfate proteoglycan
perlecan and increases in VEGF and VEGF receptor II were
prevented in mice lacking PKCa, which also had a significant
reduction in albuminuria.57 It is likely that PKC activation
has an important role in the thickening of basement
membranes as the use of PKCb selective isoforms inhibitor,
ruboxistaurin (RBX) was able to prevent both mesangial
expansion and basement membrane thickening in diabetic
db/db mice and hypertensive diabetic rats.58,59
ISOFORM SELECTIVITY AND PKC INHIBITION IN DIABETIC
NEPHROPATHY
Various isoforms have been reported to be changed by
diabetes or high glucose in vascular cells. Immunoblotting
studies have demonstrated that PKCa and PKCb1 were
increased in membranous fraction of diabetic rat glomeruli
and in mesangial cells exposed to high glucose,50 whereas
PKCb2 was reported to be preferentially activated in the aorta
and heart of diabetic rats.17 PKCb2 and PKCd have been
demonstrated to be activated in aortic vascular smooth
muscle cells grown in high glucose concentration,60 whereas
other isoforms have been observed in the retina (a, b1, b2,
and e).18 Whiteside and Dlugosz61 suggested that other PKC
isoforms, including PKCd and PKCz, may also be important
S51

in diabetic nephropathy. The function of individual PKC
isoforms is likely conferred, in part, by their subcellular
localization and binding to specific anchoring proteins after
activation and translocation.62,63
The specific effects of the PKC isoforms on the renal and
other vascular tissues to induce diabetic complications are
just beginning to be determined using pharmacological
approaches and genetically altered mice. Menne et al.57
has reported that diabetic PKCa-isoform-null mice was
resistant to albuminuria and glomerular upregulation of
VEGF and its receptor expression. Further, we have recently
reported that lack of PKCb can protect against diabetes-
induced renal hypertrophy, glomerular hyperfiltration, and
increased production of pro-fibrotic growth factors and
reactive oxygen species using PKCb-null mice.51 With the
availability of selective PKCb inhibitor (RBX), the role of
PKCb activation has been extensively studied in the area of
diabetic vascular complications. Treatment of diabetic
animals with RBX was associated with normalization of
hemodynamic changes, ECM accumulation, and histological
features of glomerular damage associated with dia-
betes.15,50,58 The preventive actions of RBX on diabetes-
induced renal pathologies have been reported in several
animal models of diabetes, including both chemically
induced and genetically programmed mice and rats in
chronic studies lasting 6 months or longer.
Owing to the large amount pre-clinical data that showed
RBX can prevent retinal and renal pathologies with very little
evidence of toxicity, RBX has been used in several clinical
trials to determine its efficacy in diabetic macular edema,
retinopathy, neuropathy, nephropathy, and endothelial dys-
function in types I and II diabetic patients. Successful results
in Phase 3 trial for diabetic macular edema has been
reported.64 Results of Phase 2 clinical trial for diabetic
proteinuria suggested that RBX can decrease the loss of
glomerular filtration rate and proteinuria in diabetic patients
who are already treated optimally with angiotensin II
inhibitors or receptor blockers.65 Phase 3 clinical trials in
diabetic nephropathy are urgently needed to confirm the
positive results from the Phase 2 study.
CONCLUSION
Currently available therapeutic approaches for diabetic ne-
phropathy are focused on intensive blood glucose control and
blockade of reninangiotensin system. Over the next few years,
we should have several new therapeutic compounds that are
derived from a basic understanding of the mechanisms by
which diabetes causes vascular complications. Strategies to
target DAG-PKC pathway using isoform-specific inhibitors
could be one of the promising therapeutic options, but definite
well-designed large and long-term clinical studies are needed to
establish its efficacy for prevention and treatment of nephro-
pathy and other vascular complications in diabetic patients.
DISCLOSURE
The authors have stated they have nothing to disclose.
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H Noh and GL King: Role of PKC activation in diabetic nephropathy
REFERENCES
1. Collins AJ, Kasiske B, Herzog C et al. United States Renal Data System
2006 Annual Data Report. American Journal of Kidney Diseases 2007; 49:
S1S234.
2. Sarnak MJ, Levey AS, Schoolwerth AC et al. Kidney disease as a risk factor
for development of cardiovascular disease: a statement from the
American Heart Association Councils on kidney in cardiovascular disease,
high blood pressure research, clinical cardiology, and epidemiology and
prevention. Circulation 2003; 108: 21542169.
3. Ismail N, Becker B, Strzelczyk P et al. Renal disease and hypertension in
non-insulin-dependent diabetes mellitus. Kidney int 1999; 55: 128.
4. Parving HH, Hommel E, Mathiesen E et al. Prevalence of
microalbuminuria, arterial hypertension, retinopathy and neuropathy in
patients with insulin dependent diabetes. Br Med J (Clin Res Ed) 1988; 296:
156160.
5. The Diabetes Control Complications Trial Research G. The effect of
intensive treatment of diabetes on the development and progression
of long-term complications in insulin-dependent diabetes mellitus. N Engl
J Med 1993; 329: 977986.
6. UK Prospective Diabetes Study G. Intensive blood-glucose control with
sulphonylureas or insulin compared with conventional treatment and risk
of complications in patients with type 2 diabetes (UKPDS 33). Lancet
1998; 352: 837853.
7. Koya D, King GL. Protein kinase C activation and the development of
diabetic complications. Diabetes 1998; 47: 859866.
8. Obrosova IG, Minchenko AG, Vasupuram R et al. Aldose reductase
inhibitor fidarestat prevents retinal oxidative stress and vascular
endothelial growth factor overexpression in streptozotocin-diabetic rats.
Diabetes 2003; 52: 864871.
9. Brownlee M. Biochemistry and molecular cell biology of diabetic
complications. Nature 2001; 414: 813820.
10. Wendt T, Harja E, Bucciarelli L et al. RAGE modulates vascular
inflammation and atherosclerosis in a murine model of type 2 diabetes.
Atherosclerosis 2006; 185: 7077.
11. Nishizuka Y. Intracellular signaling by hydrolysis of phospholipids and
activation of protein kinase C. Science 1992; 258: 607614.
12. Nishizuka Y. Protein kinase C and lipid signaling for sustained cellular
responses. FASEB J 1995; 9: 484496.
13. Sheetz MJ, King GL. Molecular understanding of hyperglycemias adverse
effects for diabetic complications. JAMA 2002; 288: 25792588.
14. Derubertis FR, Craven PA. Activation of protein kinase C in glomerular
cells in diabetes. Mechanisms and potential links to the pathogenesis of
diabetic glomerulopathy. Diabetes 1994; 43: 18.
15. Ishii H, Jirousek MR, Koya D et al. Amelioration of vascular dysfunctions in
diabetic rats by an oral PKC b inhibitor. Science 1996; 272: 728731.
16. Craven PA, Davidson CM, DeRubertis FR. Increase in diacylglycerol mass
in isolated glomeruli by glucose from de novo synthesis of glycerolipids.
Diabetes 1990; 39: 667674.
17. Inoguchi T, Battan R, Handler E et al. Preferential elevation of protein
kinase C isoform {beta}II and diacylglycerol levels in the aorta and heart
of diabetic rats: differential reversibility to glycemic control by islet cell
transplantation. Proc Natl Acad Sci USA 1992; 89: 1105911063.
18. Shiba T, Inoguchi T, Sportsman JR et al. Correlation of diacylglycerol level
and protein kinase C activity in rat retina to retinal circulation. Am J
Physiol Endocrinol Metab 1993; 265: E783E793.
19. Considine RV, Nyce MR, Allen LE et al. Protein kinase C is increased in the
liver of humans and rats with non-insulin-dependent diabetes mellitus:
an alteration not due to hyperglycemia. J Clin Invest 1995; 95: 29382944.
20. Saha AK, Kurowski TG, Colca JR et al. Lipid abnormalities in tissues of the
KKAy mouse: effects of pioglitazone on malonyl-CoA and diacylglycerol.
Am J Physiol Endocrinol Metab 1994; 267: E95E101.
21. Ayo SH, Radnik R, Garoni JA et al. High glucose increases diacylglycerol
mass and activates protein kinase C in mesangial cell cultures. Am J
Physiol Renal Physiol 1991; 261: F571F577.
22. Studer RK, Craven PA, DeRubertis FR. Role for protein kinase C in the
mediation of increased fibronectin accumulation by mesangial cells
grown in high-glucose medium. Diabetes 1993; 42: 118126.
23. Cooper ME. Interaction of metabolic and haemodynamic factors in
mediating experimental diabetic nephropathy. Diabetologia 2001; 44:
19571972.
24. Christiansen JS, Gammelgaard J, Frandsen M et al. Increased kidney size,
glomerular filtration rate and renal plasma flow in short-term insulin-
dependent diabetics. Diabetologia 1981; 20: 451456.
25. Ditzel J, Schwartz M. Abnormally increased glomerular filtration
rate in short-term insulin-treated diabetic subjects. Diabetes 1967; 16:
264267.
Kidney International (2007) 72, S49S53
H Noh and GL King: Role of PKC activation in diabetic nephropathy
26. Hostetter TH, Troy JL, Brenner BM. Glomerular hemodynamics in
experimental diabetes mellitus. Kidney Int 1981; 19: 410415.
27. Viberti GC. Early functional and morphological changes in diabetic
nephropathy. Clin Nephrol 1979; 12: 4753.
28. ODonnell MP, Kasiske BL, Keane WF. Glomerular hemodynamic and
structural alterations in experimental diabetes mellitus. FASEB J 1988; 2:
23392347.
29. Ruan X, Arendshorst WJ. Role of protein kinase C in angiotensin
II-induced renal vasoconstriction in genetically hypertensive rats.
Am J Physiol Renal Physiol 1996; 270: F945F952.
30. Craven PA, Caines MA, DeRubertis FR. Sequential alterations in
glomerular prostaglandin and thromboxane synthesis in diabetic rats:
relationship to the hyperfiltration of early diabetes. Metabolism 1987; 36:
95103.
31. Perico N, Benigni A, Gabanelli M et al. Atrial natriuretic peptide and
prostacyclin synergistically mediate hyperfiltration and hyperperfusion of
diabetic rats. Diabetes 1992; 41: 533538.
32. Lin L-L, Wartmann M, Lin AY et al. cPLA2 is phosphorylated and activated
by MAP kinase. Cell 1993; 72: 269278.
33. Williams B, Schrier RW. Glucose-induced protein kinase C activity
regulates arachidonic acid release and eicosanoid production by cultured
glomerular mesangial cells. J Clin Invest 1993; 92: 28892896.
34. Xia P, Kramer RM, King GL. Identification of the mechanism for the
inhibition of Na+,K(+)-adenosine triphosphatase by hyperglycemia
involving activation of protein kinase C and cytosolic phospholipase A2.
J Clin Invest 1995; 96: 733740.
35. Haneda M, Araki S, Togawa M et al. Mitogen-activated protein kinase
cascade is activated in glomeruli of diabetic rats and glomerular
mesangial cells cultured under high glucose conditions. Diabetes 1997;
46: 847853.
36. Bank N, Aynedjian HS. Role of EDRF (nitric oxide) in diabetic renal
hyperfiltration. Kidney Int 1993; 43: 13061312.
37. Tolins JP, Shultz PJ, Raij L et al. Abnormal renal hemodynamic response to
reduced renal perfusion pressure in diabetic rats: role of NO. Am J Physiol
Renal Physiol 1993; 265: F886F895.
38. Komers R, Allen TJ, Cooper ME. Role of endothelium-derived nitric oxide
in the pathogenesis of the renal hemodynamic changes of experimental
diabetes. Diabetes 1994; 43: 11901197.
39. Sharma K, Danoff TM, Depiero A et al. Enhanced expression of inducible
nitric oxide synthase in murine macrophages and glomerular mesangial
cells by elevated glucose levels: possible mediation via protein kinase C.
Biochem Biophys Res Commun 1995; 207: 8088.
40. Craven PA, Studer RK, DeRubertis FR. Impaired nitric oxide-dependent
cyclic guanosine monophosphate generation in glomeruli from diabetic
rats. Evidence for protein kinase C-mediated suppression of the
cholinergic response. J Clin Invest 1994; 93: 311320.
41. Cooper ME, Vranes D, Youssef S et al. Increased renal expression of
vascular endothelial growth factor (VEGF) and its receptor VEGFR-2 in
experimental diabetes. Diabetes 1999; 48: 22292239.
42. Chou E, Suzuma I, Way KJ et al. Decreased cardiac expression of vascular
endothelial growth factor and its receptors in insulin-resistant and
diabetic states: a possible explanation for impaired collateral formation in
cardiac tissue. Circulation 2002; 105: 373379.
43. Aiello LP, Bursell SE, Clermont A et al. Vascular endothelial growth factor-
induced retinal permeability is mediated by protein kinase C in vivo and
suppressed by an orally effective beta-isoform-selective inhibitor.
Diabetes 1997; 46: 14731480.
44. Haneda M, Kikkawa R, Horide N et al. Glucose enhances type IV collagen
production in cultured rat glomerular mesangial cells. Diabetologia 1991;
34: 198200.
45. Fumo P, Kuncio GS, Ziyadeh FN. PKC and high glucose stimulate collagen
alpha 1 (IV) transcriptional activity in a reporter mesangial cell line. Am J
Physiol Renal Physiol 1994; 267: F632F638.
Kidney International (2007) 72, S49S53
46. Sharma K, Ziyadeh FN. Hyperglycemia and diabetic kidney disease. The
case for transforming growth factor-beta as a key mediator. Diabetes
1995; 44: 11391146.
47. Yamamoto T, Nakamura T, Noble NA et al. Expression of transforming
growth factor {beta} is elevated in human and experimental diabetic
nephropathy. Proc Natl Acad Sci USA 1993; 90: 18141818.
48. Shankland SJ, Scholey JW, Ly H et al. Expression of transforming growth
factor-beta 1 during diabetic renal hypertrophy. Kidney Int 1994; 46:
430442.
49. Ziyadeh FN, Sharma K, Ericksen M et al. Stimulation of collagen gene
expression and protein synthesis in murine mesangial cells by high
glucose is mediated by autocrine activation of transforming growth
factor-beta. J Clin Invest 1994; 93: 536542.
50. Koya D, Jirousek MR, Lin Y-W et al. Characterization of protein
kinase C beta isoform activation on the gene expression of
transforming growth factor-beta, extracellular matrix components,
and prostanoids in the glomeruli of diabetic rats. J Clin Invest 1997; 100:
115126.
51. Ohshiro Y, Ma RC, Yasuda Y et al. Reduction of diabetes-induced oxidative
stress, fibrotic cytokine expression, and renal dysfunction in protein
kinase c{beta}-null mice. Diabetes 2006; 55: 31123120.
52. Oliver JA. Adenylate cyclase and protein kinase C mediate
opposite actions on endothelial junctions. J Cell Physiol 1990; 145:
536542.
53. Lynch JJ, Ferro TJ, Blumenstock FA et al. Increased endothelial albumin
permeability mediated by protein kinase C activation. J Clin Invest 1990;
85: 19911998.
54. Werth DK, Niedel JE, Pastan I. Vinculin, a cytoskeletal substrate of protein
kinase C. J Biol Chem 1983; 258: 1142311426.
55. Stasek Jr JE, Patterson CE, Garcia JG. Protein kinase C phosphorylates
caldesmon77 and vimentin and enhances albumin permeability across
cultured bovine pulmonary artery endothelial cell monolayers. J Cell
Physiol 1992; 153: 6275.
56. Xia P, Aiello LP, Ishii H et al. Characterization of vascular endothelial
growth factors effect on the activation of protein kinase C, its isoforms,
and endothelial cell growth. J Clin Invest 1996; 98: 20182026.
57. Menne J, Park J-K, Boehne M et al. Diminished loss of proteoglycans and
lack of albuminuria in protein kinase c-{alpha} deficient diabetic mice.
Diabetes 2004; 53: 21012109.
58. Koya D, Haneda M, Nakagawa H et al. Amelioration of accelerated
diabetic mesangial expansion by treatment with a PKC {beta} inhibitor in
diabetic db/db mice, a rodent model for type 2 diabetes. FASEB J 2000;
14: 439447.
59. Kelly DJ, Zhang Y, Hepper C et al. Protein kinase C {beta} inhibition
attenuates the progression of experimental diabetic nephropathy in the
presence of continued hypertension. Diabetes 2003; 52: 512518.
60. Igarashi M, Wakasaki H, Takahara N et al. Glucose or diabetes activates
p38 mitogen-activated protein kinase via different pathways. J Clin Invest
1999; 103: 185195.
61. Whiteside CI, Dlugosz JA. Mesangial cell protein kinase C isozyme
activation in the diabetic milieu. Am J Physiol Renal Physiol 2002; 282:
F975F980.
62. Csukai M, Mochly-Rosen D. Pharmacologic modulation of protein kinase
C isozymes: the role of racks and subcellular localisation. Pharmacol Res
1999; 39: 253259.
63. Mochly-Rosen D, Gordon AS. Anchoring proteins for protein kinase C: a
means for isozyme selectivity. FASEB J 1998; 12: 3542.
64. PKC-DRS2 GroupAiello L, Davis M et al. Effect of ruboxistaurin on visual
Loss in patients with diabetic retinopathy. Ophthalmology 2006; 113:
22212230.
65. Tuttle KR, Bakris GL, Toto RD et al. The effect of ruboxistaurin
on nephropathy in type 2 diabetes. Diabetes Care 2005; 28:
26862690.
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