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Am J Physiol Heart Circ Physiol 294: H2785-H2791, 2008. First published April 25, 2008; doi:10.1152/ajpheart.00149.2008
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Statins restore ischemic limb blood flow in diabetic microangiopathy via eNOS/NO upregulation but not via PDGF-BB expression

Takaaki Fujii,1,3 Mitsuho Onimaru,1 Yoshikazu Yonemitsu,2 Hiroyuki Kuwano,3 and Katsuo Sueishi1

1Division of Pathophysiological and Experimental Pathology, Department of Pathology, Graduate School of Medical Science, Kyushu University, Fukuoka; 2Department of Gene Therapy, Graduate School of Medicine, Chiba University, Chiba; and 3Department of General Surgical Science, Graduate School of Medicine, Gunma University, Gunma, Japan

Submitted 12 February 2008 ; accepted in final form 11 April 2008


   ABSTRACT
TOP
 ABSTRACT
MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 GRANTS
 REFERENCES
 
3-Hydroxy-3-methyl-glutaryl CoA reductase inhibitors, or statins, have pleiotropic effects and can protect the vasculature in a manner independent of their lipid-lowering effect. The effectiveness of statins in reducing the risk of coronary events has been shown even in patients with diabetes, and their effects on diabetic complications have been reported. Using a model of severe hindlimb ischemia in streptozotocin-induced diabetic mice (STZ-DM), we investigated the effects and mechanisms of statin therapy in diabetic angiopathy in ischemic hindlimbs. As a result, STZ-DM mice frequently lost their hindlimbs after induced ischemia, whereas non-DM mice did not. Supplementation with statins significantly prevented autoamputation. We previously showed that diabetic vascular complications are caused by impaired expression of PDGF-BB, but statin therapy did not enhance PDGF-BB expression. Statins helped enhance endogenous endothelial nitric oxide (NO) synthase (eNOS) expression. Furthermore, the inhibition of NO synthesis by the administration of N{omega}-nitro-L-arginine methyl ester impaired the ability of statins to prevent STZ-DM mouse limb autoamputation, indicating that the therapeutic effect of statins in hindlimb ischemia in STZ-DM mice occurs via the eNOS/NO pathway. A combination therapy of statins and PDGF-BB gene supplementation was more effective for diabetic angiopathy than either therapy alone. In conclusion, these findings indicate that statin therapy might be useful for preventing intractable diabetic foot disease in patients with diabetic angiopathy.

endothelial nitric oxide synthase; nitric oxide; platelet-derived growth factor-BB; diabetes mellitus

DIABETES MELLITUS (DM) is characterized by a chronic state of hyperglycemia and is increasing to epidemic proportions throughout the world. The morbidity and mortality associated with diabetes is primarily due to macro- and microangiopathy occurring in multiple organs (2, 20). The diabetic foot is an intractable disease categorized by DM-related vascular complications, and patients with it have a much higher risk of gangrene and the need for consequent amputation of the lower extremities (20). Collateral vessel development is insufficient to compensate with the reduced blood flow through occluded arteries in patients with peripheral vascular disease, especially DM (10). Furthermore, surgical and catheter interventions are usually difficult to treat limb ischemia of DM patients with DM, because vascular diseases are located in small vessels.

3-Hydroxy-3-methyl-glutaryl CoA reductase inhibitors or statins are potent inhibitors of cholesterol biosynthesis that have become more widely used in greater numbers of patients with hypercholesterolemia (13, 31). Furthermore, recent studies suggest that statins have pleiotropic effects in a manner independent of their lipid-lowering effect and can protect the vasculature (8, 27, 29). Several trials have demonstrated the beneficial effects of statins in lowering cardiovascular-related morbidity and mortality in patients with coronary artery disease. The effectiveness of statins in reducing the risk of coronary events has been established in patients with and without diabetes, and it has been suggested that patients with diabetes benefit more than patients without in both primary and secondary prevention (6).

Recent studies have also revealed the modulatory effects of statins in diabetic microangiopathy (7). The effectiveness of statins appears to involve restoring or improving endothelial function through the attenuation of high glucose-induced or diabetes-induced oxidative stress, thereby increasing the bioavailability of nitric oxide (NO) or inhibiting inflammatory responses (7).

On the other hand, using a model of severe hindlimb ischemia in streptozotocin-induced diabetic mice (STZ-DM), we previously showed that the diabetic foot is a disease involving the disturbance of PDGF-BB expression but not of the disturbance of the responses of angiogenic factors (32). Screening of angiogenesis-related factors revealed that the expression of PDGF-BB was impaired in STZ-DM mice at baseline as well as over a time course after limb ischemia. Increased expression of PDGF-BB prevented autoamputation in the STZ-DM mice (32).

In this study, we investigated the effect of and the mechanism underlying statin therapy in hindlimb ischemia under chronic hyperglycemia using STZ-DM. We here demonstrate that statins show significant therapeutic effects in hindlimb ischemia in STZ-DM mice via the endothelial NO synthase (eNOS)/NO pathway but not via PDGF-BB expression. Our results suggest that statin therapy would be useful for preventing intractable diabetic foot disease in patients with diabetic angiopathy.


   MATERIALS AND METHODS
TOP
 ABSTRACT
 MATERIALS AND METHODS
RESULTS
 DISCUSSION
 GRANTS
 REFERENCES
 
Cells and reagents. Human umbilical vascular endothelial cells (HUVECs) were purchased from Kurabo, Tokyo, Japan. Two intracellular signal inhibitors were used as previously described (11, 12, 23, 32, 33): bis-I (a PKC inhibitor) and its inactive control, bis-V (100 mmol/l, Sigma-Aldrich, Tokyo, Japan).

Animals. Male C57BL/6J (7 wk old) were purchased from KBT Oriental (Charles River Grade, Tosu, Saga, Japan). All animal experiments were performed according to approved protocols and in accordance with recommendations for the proper care and use of laboratory animals by the Committee for Animals, Recombinant DNA, and Experiments Using Infectious Pathogens at Kyushu University, and according to Japanese government Law No. 105 and Notification No. 6. Experimental diabetes was induced in mice by daily intravenous injection of STZ in citrate buffer (1.5 mg/body) for 5 days (days –5 to 0) for the Type 1 diabetic model. As for Type 2 diabetic models, 10-wk-old male ob/ob (C57BL/6J-Lepob/Lepob) mice (4) and normal control homozygous (+/+) mice (KBT Oriental) were also used to confirm eNOS and PDGF-BB expression. For statin therapy, pravastatin and pitavastatin (0.2 mg/g body wt; injected volume, 0.02 ml/g body wt) were administered by intraperitoneal injection during days 14 to 28. To inhibit NO synthesis, N{omega}-nitro-L-arginine methyl ester (1 mg/ml in drinking water; Sigma) was orally administered during days 14 to 28.

Murine severe hindlimb ischemia. Details of the surgical treatment and evaluation of limb prognosis have been described previously (11, 12, 18, 23, 32, 33); specifically, the excision of both the left femoral artery and vein and their branches from the inguinal ligament up to and including the saphenous-popliteal bifurcation was performed.

Gene transfer vectors. The plasmid-based gene of human PDGF-BB was prepared as previously described (32). Human full-length cDNA of PDGF-B (GenBank: No. BC029822) was amplified by PCR using specific primers (forward: 5'-AAGGTACCATGAATCGCTGCTGGGCGCTC-3' and reverse: 5'-TTCTCGAGCTAGGCTCCAAGGGTCTCCTTC-3') and subcloned into a TA cloning vector (Invitrogen, San Diego, CA). The whole sequence was then determined using the CEQ 2000 Sequence Detection System (Beckman Coulter, Fullerton, CA). The amplicon was transferred to the KpnI-XhoI sites of the mammalian expression vector pCEP4 (Invitrogen).

ELISA. The protein contents in the limb muscles and culture medium were determined using Quantikine Immunoassay systems for human eNOS (available for both humans and mice: R&D Systems, Minneapolis, MN), murine PDGF-BB (R&D Systems) and human PDGF-BB (specific for humans: R&D Systems) according to the manufacturer's instructions, as previously described (11, 12, 18, 23, 32, 33).

Biochemical analysis. Serum levels of LDL cholesterol and advanced glycation end product (AGE) were measured in serum samples when all animals were euthanized. Concentrations of LDL cholesterol and AGE were determined by an automatic analyzer (Research Testing Department, SRL, Hachiohji-shi, Tokyo, Japan).

Laser-Doppler perfusion images. Measurements of the ischemic (left)/normal (right) limb blood flow ratio were made using a laser-Doppler perfusion image (LDPI) analyzer (Moor Instruments, Devon, UK) as previously described (11, 12, 18, 23, 32, 33). To minimize data variability due to ambient light and temperature, the LDPI index was expressed as the ratio of the left (ischemic) to the right (nonischemic) limb blood flow.

Statistical analysis. All data except for those of limb survival were expressed as means ± SE and were analyzed by one-way ANOVA with Fisher adjustment. For the survival analysis, the survival rate expressed by the limb salvage score was analyzed using Kaplan-Meyer's method (11, 18, 23, 32, 33). The statistical significance of the survival experiments was determined using the log-rank test. P < 0.05 was considered statistically significant in all analyses.


   RESULTS
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 ABSTRACT
 MATERIALS AND METHODS
 RESULTS
DISCUSSION
 GRANTS
 REFERENCES
 
Effects of statins on serum glucose, LDL cholesterol, and AGE in DM mice. Serum levels of glucose, LDL cholesterol (LDL), and AGE were measured in serum samples of a relevant model for Type 1 diabetes, STZ-DM mice, and a well-accepted model of Type 2 diabetes, namely ob/ob mice that are leptin-deficient C57BL/6 (4). Serum concentrations of glucose, LDL, and AGE were increased in the DM mice. Statin treatment did not affect the increases in glucose and AGE, whereas the increased LDL concentration was relatively decreased (Table 1). In the following experiments, STZ-DM mice were used after we confirmed a significant upregulation of the serum glucose.


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  		Table 1. Serum LDL cholesterol, AGE, and glucose concentration are significantly increased in C57BL/6J strain-based 10-week-old Type 1 (STZ-DM) and Type 2 (Lepob/Lepob: ob/ob) mice

 
Therapeutic effects of statins in hindlimb ischemia in STZ-DM mice. The STZ-DM mice frequently lost their hindlimbs at various levels after surgically induced severe limb ischemia, although the non-DM mice did not. Quantitative analysis of the degree of autoamputation using the limb salvage score (11, 18, 23, 32, 33) demonstrated impaired limb survival in the STZ-DM mice. Administration of statins (pravastatin and pitavastatin) by intraperitoneal injection daily on days 14 to 38 resulted in the significant prevention of autoamputation in the STZ-DM mice (Fig. 1), indicating that the statin therapy effectively restored tolerance against hindlimb ischemia in STZ-DM mice.


Figure 1
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  		Fig. 1. Limb prognosis curve according to the limb salvage score in streptozotocin-induced diabetes (STZ-DM) C57/BL6 mice. These curves were obtained using Kaplan-Meyer's method, and data were analyzed using the log-rank test. Administration of statins, pravastatin (PRA, hydrophilic statin) or pitavastatin (PTA, lipophilic statin), by daily intraperitoneal injection on days 14 to 38, resulted in the significant prevention of autoamputation in the STZ-DM mice (n = animals/group).

 
eNOS/NO is important for the therapeutic effects of statins, but endogenous PDGF-BB expression is not. We previously showed that the diabetic foot is a disease involving the disturbance of PDGF-BB expression (32). To explain the mechanism underlying the therapeutic effect of statins on the tolerance of limb ischemia in STZ-DM mice, we measured the expression of murine (m)PDGF-BB protein in thigh muscles of STZ-DM. Downregulated expression of PDGF-BB was evident in both the STZ-DM, and statin therapy did not influence the impaired expression of PDGF-BB (Fig. 2A). In addition, the expression of mPDGF-BB protein was not influenced by statin therapy even in ob/ob mice, a model of Type 2 diabetes. To confirm the modulatory effect of statins on PDGF-BB expression, we examined the induction of PDGF-BB via statins using cultured HUVECs. As shown in Fig. 2B, the statins did not stimulate the production or secretion of PDGF-BB in the culture medium of HUVECs. These results suggest that the effectiveness of statins in hindlimb ischemia in DM mice may be independent of the endogenous expression of PDGF-BB.


Figure 2
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  		Fig. 2. A: comparison of protein expression of murine PDGF-BB in the thigh muscles of non-DM or DM mice assessed by ELISA. Impaired expression of murine PDGF-BB (mPDGF-BB) protein in C57BL/6J strain-based 10-wk-old Type 1 (STZ-DM; left) and Type 2 (Lepob/Lepob: ob/ob; right) mice was not restored by statin treatment (n = 6 animals/group). *P < 0.01; #P < 0.05 vs. non-DM. B: statins did not contribute to the PDGF-BB expression in human umbilical vascular endothelial cells (HUVECs). Twenty-four hours after preincubation with 5% FBS, HUVECs were stimulated with statins (0.1 or 1 mM, respectively). Twenty-four hours later, the culture medium was subjected to ELISA. BIS-I, PKC inhibitor; BIS-V, PKC inhibitor control compound; Cont, control; hPDGF-BB, human PDGF-BB. *P < 0.01.

 
We previously reported that PDGF-B gene expression was downregulated in the limb muscles of STZ-DM mice among the factors tested (VEGF-A and -C, hepatocyte growth factor, FGF-2, PDGF-A and -B, and angiopoietin-1 and -2), as well as their receptors (tie-2, flk-1, fibroblast growth factor receptor 1, flt-4, and platelet-derived growth factor receptor-A and -B) (32). Recent important studies indicated that statins activated and upregulated the expression of endogenous eNOS, which has angiogenic potency (1, 21). We next investigated the expression of eNOS and whether or not eNOS/NO is important for the ability of statins to prevent the impairment of limb survival. Statin therapy significantly restored or increased the expression of eNOS in thigh muscles of both STZ-DM and ob/ob DM mice (Fig. 3A). As in previous studies (17, 19, 22, 24, 34), statin stimuli induced the upregulation of endogenous eNOS in the lysate of the cultured cells (HUVECs). Furthermore, the daily oral administration of N{omega}-nitro-L-arginine methyl ester, an inhibitor of NO synthesis, prevented the autoamputation rate for ischemic limb amputation of STZ-DM mice (Fig. 4), indicating that the therapeutic effect of statins on tolerance against limb ischemia in the diabetic foot might occur at least partly via the eNOS/NO pathway.


Figure 3
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  		Fig. 3. A: endothelial nitric oxide (NO) synthase (eNOS) protein expression in the thigh muscles of non-DM or DM mice assessed by ELISA. Statin therapy significantly restored or increased the expression of eNOS in both STZ-DM (right) and ob/ob (left) DM mice (n = 6 animals/group). *P < 0.01; #P < 0.05 vs. non-DM. B: eNOS expression was increased by statin stimulation in HUVECs. Twenty-four hours after preincubation with 5% FBS, HUVECs were stimulated with statins (0.01 or 0.1 mM, respectively). Twenty-four hours later, the lysate of the cultured HUVECs was subjected to ELISA. *P < 0.01; #P < 0.05.

 

Figure 4
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  		Fig. 4. Limb prognosis curve according to the limb salvage score after the daily oral administration of N{omega}-nitro-L-arginine methyl ester (L-NAME), an inhibitor of NO synthesis in STZ-DM mice. These curves were obtained using Kaplan-Meyer's method, and the data were analyzed using the log-rank test. Administration of L-NAME prevented the therapeutic effect for ischemic limb autoamputation in STZ-DM mice by treatment with either statin: PRA (left) and PTA (right). #P < 0.05 vs. others.

 
Combination therapy of statins and PDGF-BB gene supplementation for diabetic angiopathy. These results may suggest that the therapeutic effect of statins occurs via eNOS/NO but not via PDGF-BB expression in diabetic vascular dysfunction. As a final assessment, to confirm whether or not statin therapy is independent of the endogenous expression of PDGF-BB, we administered a combination therapy of statins and PDGF-BB gene supplementation and found that this combination is more effective for diabetic angiopathy than either treatment alone. A supplementation study on the plasmid-based intramuscular gene transfer of human PDGF-B (pCEP4-hPDGFB) was previously described (32). In the present study we assessed the recovery of blood flow evaluated by LDPI, because the PDGF-B gene transfer resulted in the complete prevention of autoamputation in STZ-DM mice. As shown in Fig. 5, both statin therapy and the preinjection of pCEP4-hPDGFB significantly improved the disturbed blood perfusion in STZ-DM mice; furthermore, the combination therapy had an effect on diabetic angiopathy that was superior to that by either therapy alone, indicating that the statin therapy might work independently from PDGF-BB expression and that the combination therapy was sufficient to restore the tolerance against hindlimb ischemia in STZ-DM mice.


Figure 5
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  		Fig. 5. Effects of statin therapy and/or plasmid-based PDGF-B gene transfer on recovery of blood flow in ischemic limbs of STZ-DM mice. Administration of PRA or PTA by daily intraperitoneal injection on days 14 to 38 significantly prevented autoamputation in the STZ-DM mice. pCEP4-hPDGFB and control plasmid (pCEP4-empty) were injected into the thigh muscle 2 days before induced limb ischemia. After assessment of blood flow before or after ischemic operation, recovery of blood flow was assessed by laser-Doppler perfusion imaging at each time point. Data were standardized by data related to the untreated right limb and expressed as the ratio of flow intensity (in %). *P < 0.01; #P < 0.05.

 

   DISCUSSION
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 ABSTRACT
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
GRANTS
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The key observations made in this study are summarized as follows: 1) the administration of statin was effective for preventing autoamputation of the ischemic limb of STZ-DM mice; 2) statin therapy could not restore the disturbed expression of PDGF-BB in DM mice, suggesting that the effect of statins is independent of PDGF-BB expression; 3) statins could restore or increase eNOS expression in the limb of DM mice; 4) supplementation with the NO inhibitor inhibited the ability of statins to prevent autoamputation, indicating that the therapeutic effect of statins in hindlimb ischemia in STZ-DM mice might occur via the eNOS/NO pathway; and 5) the combination therapy of statins and PDGF-BB supplementation was more effective than either therapy alone. These findings suggest that the statin therapy is effective against the formation of hyperglycemia-related vascular complications, and they imply that statin therapy improves limb survival and perfusion via a mechanism involving eNOS/NO pathway in Type 1 DM, and probably in Type 2 DM microangiopathy.

Over the last several years, it has been already demonstrated that statin treatment improved and promoted angiogenesis in a hindlimb ischemia of non-DM mouse model (16, 26). In the case of DM mouse model, we previously demonstrated that the disturbed tolerance against severe limb ischemia under hyperglycemia was due to the disturbance of PDGF-BB expression and not to the angiogenic responses and that the supplementation of PDGF-B gene expression was sufficient to prevent autoamputation due to limb ischemia in STZ-DM mice (32). The reduction of PDGF-BB expression, which was not dependent on the level of hyperglycemia (32), was critical to inducing functional and morphological vascular change, which is the dissociation of pericytes from the capillaries in muscles of STZ-DM mice, indicating that impaired PDGF-BB expression disturbed vessel maturation. In the present study, statin administration did not influence PDGF-BB expression, which is concerned with vessel maturation. We postulated that the mechanism underlying the therapeutic effect of statins might be due to improved endothelial function because of the lower PDGF-BB expression. Many diabetic vascular complications involve endothelial cell dysfunction characterized by reduced NO-dependent phenomena, including vasodilation and protection against leucocyte-endothelial interactions. Several studies have shown that hyperglycemia impairs NO production and have demonstrated impaired endothelium-dependent vasorelaxation in diabetic humans and in experimental diabetic animals (5, 14, 15, 30). A hallmark of endothelial dysfunction is reduced bioavailability of NO, which could be caused by reduced expression of eNOS, impairment of eNOS activation, or increased inactivation of NO by oxidative stress. Upregulation of the activity or expression of eNOS is considered to be effective in diabetic angiopathy, and statins can increase eNOS expression and activation in addition to their lipid-lowering effect (8, 17, 19, 22, 24, 27, 34). Furthermore, statins have also been reported to promote angiogenesis via eNOS (16, 26). In our current study, statin therapy was effective for preventing autoamputation of the ischemic limb under hyperglycemia at least partly via eNOS, and those previous reports essentially support our findings that statins could restore or increase the eNOS expression in the ischemic limb of DM mice.

In addition to eNOS/NO dependent pathway, it has been revealed that statins have additional effects of growing interest that include the ability to recruit endothelial progenitor cells (3, 8, 9) or activate the protein kinase Akt, which leads to angiogenesis and prevents apoptosis in endothelial cells (9,16). As shown Fig. 4, our current findings indicate that the therapeutic effects of statins on tolerance against limb ischemia in the diabetic foot might occur at least partly via the eNOS/NO pathway. Therefore, there is a possibility that the clinical benefits of statin therapy might be caused partly by these NO-independent pathways; however, further extensive studies should be carried out to determine this hypothesis.

Although statins have the favorable effect of restoring blood flow in the ischemic limb of DM mice (16, 26), they prevented autoamputation of the ischemic limb of STZ-DM mice to some extent, but not completely. In turn, this result also suggests that the therapeutic effect of statins is due to improved endothelial function and is independent of the restoration of the impairment PDGF-BB expression, which was sufficient to prevent autoamputation due to limb ischemia in STZ-DM mice. Furthermore, the results of the combination therapy of statins and PDGF-BB supplementation confirmed that the favorable effect of statin therapy on the diabetic foot occurs by mechanisms other than the upregulation of PDGF-BB expression.

Some clinical trials have suggested that the vasculature effects of hydrophilic statins are similar in extent to those of lipophilic statins, but it is still controversial whether lipophilic or hydrophilic statins have more clinical benefits (23a, 25). An important advance of our current study was to determine that there are clearly differences between lipophilic and hydrophilic statins, although both types are beneficial to ischemic limbs under hyperglycemia. Limb survival was similar in mice receiving lipophilic and hydrophilic statins; however, eNOS expression was more upregulated by the administration of lipophilic statins than by that of hydrophilic statins (Fig. 3). Therefore, the present study indicates that clinically lipophilic statins may be more useful for ischemic diabetic foot.

In conclusion, we demonstrated that statin therapy restored the disturbed blood flow of severe limb ischemia under hyperglycemia by increasing eNOS/NO expression and not by increasing PDGF-BB expression and that the inhibition of NO reduced the ability of statins to prevent autoamputation due to limb ischemia in STZ-DM mice. Therefore, statin therapy is expected to be useful for preventing intractable diabetic foot disease in patients with diabetic angiopathy via eNOS/NO.


   GRANTS
TOP
 ABSTRACT
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 GRANTS
REFERENCES
 
This work was supported in part by a Japanese Ministry of Education, Culture, Sports, Science, and Technology of Japan grant-in-aid (to Y. Yonemitsu and K. Sueishi) and by the National Institute of Biomedical Innovation (Japan) Program for Promotion of Fundamental Studies in Health Sciences No. MF-21 (to Y. Yonemitsu and K. Sueishi).


   ACKNOWLEDGMENTS
 
We thank Chie Arimatsu for assistance with the animal experiments.


   FOOTNOTES
 

Address for reprint requests and other correspondence: T. Fujii, Div. of Pathophysiological and Experimental Pathology, Dept. of Pathology, Graduate School of Medical Sciences, Kyushu Univ., 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan

The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.


   REFERENCES
TOP
 ABSTRACT
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
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 REFERENCES
 

  1. Amano K, Matsubara H, Iba O, Okigaki M, Fujiyama S, Imada T, Kojima H, Nozawa Y, Kawashima S, Yokoyama M, Iwasaka T. Enhancement of ischemia-induced angiogenesis by eNOS overexpression. Hypertension 41: 156–162, 2003.[Abstract/Free Full Text]
  2. Amos A, McCarty D, Zimmet P. The rising global burden of diabetes and its complications: estimates and projections to the year 2000. Diabet Med 14: S1–S85, 1997.[Medline]
  3. Assmus B, Urbich C, Aicher A, Hofmann WK, Haendeler J, Rössig L, Spyridopoulos I, Zeiher AM, Dimmeler S. HMG-CoA reductase inhibitors reduce senescence and increase proliferation of endothelial progenitor cells via regulation of cell cycle regulatory genes. Circ Res 92: 1049–1055, 2003.[Abstract/Free Full Text]
  4. Bailey CJ, Flatt PR, Atkins TW. Influence of genetic background and age on the expression of the obese hyperglycaemic syndrome in Aston ob/ob mice. Int J Obes 6: 11–21, 1982.[Web of Science][Medline]
  5. Chakravarthy U, Hayes RG, Stitt AW, McAuley E, Archer DB. Constitutive nitric oxide synthase expression in retinal vascular endothelial cells is suppressed by high glucose and advanced glycation end products. Diabetes 47: 945–952, 1998.[Abstract]
  6. Costa J, Borges M, David C, Vaz Carneiro A. Efficacy of lipid lowering drug treatment for diabetic and non-diabetic patients: meta-analysis of randomized controlled trials. BMJ 332: 1115–1124, 2006.[Abstract/Free Full Text]
  7. Danesh FR, Kanwar YS. Modulatory effects of HMG-CoA reductase inhibitors in diabetic microangiopathy. FASEB J 18: 805–815, 2004.[Abstract/Free Full Text]
  8. Davignon J. Beneficial cardiovascular pleiotropic effects of statins. Circulation 109: III39–III43, 2004.[Medline]
  9. Dimmeler S, Aicher A, Vasa M, Mildner-Rihm C, Adler K, Tiemann M, Rütten H, Fichtlscherer S, Martin H, Zeiher AM. HMG-CoA reductase inhibitors (statins) increase endothelial progenitor cells via the PI3-kinase/Akt pathway. J Clin Invest 108: 391–397, 2001.[CrossRef][Web of Science][Medline]
  10. Feener EP, King GL. Vascular dysfunction in diabetes mellitus. Lancet 350: SI9–SI13, 1997.[Web of Science][Medline]
  11. Fujii T, Yonemitsu Y, Onimaru M, Tanii M, Nakano T, Egashira K, Takehara T, Inoue M, Hasegawa M, Kuwano H, Sueishi K. Nonendothelial mesenchymal cell-derived MCP-1 is required for FGF-2-mediated therapeutic neovascularization: critical role of the inflammatory/arteriogenic pathway. Arterioscler Thromb Vasc Biol 26: 2483–2489, 2006.[Abstract/Free Full Text]
  12. Fujii T, Yonemitsu Y, Onimaru M, Inoue M, Hasegawa M, Kuwano H, Sueishi K. VEGF function for upregulation of endogenous PlGF expression during FGF-2-mediated therapeutic angiogenesis. Atherosclerosis. In press.
  13. Gaw A. A new reality: achieving cholesterol-lowering goals in clinical practice. Atherosclerosis 2: S5–S8, 2002.
  14. Guzik TJ, Mussa S, Gastaldi D, Sadowski J, Ratnatunga C, Pillai R, Channon KM. Mechanisms of increased vascular superoxide production in human diabetes mellitus: role of NAD(P)H oxidase and endothelial nitric oxide synthase. Circulation 105: 1656–1662, 2002.[Abstract/Free Full Text]
  15. Hink U, Li H, Mollnau H, Oelze M, Matheis E, Hartmann M, Skatchkov M, Thaiss F, Stahl RAK, Warnholz A, Meinertz T, Griendling K, Harrison DG, Forstermann U, Munzel T. Mechanisms underlying endothelial dysfunction in diabetes mellitus. Circ Res 88: e14–e22, 2001.[Web of Science][Medline]
  16. Kureishi Y, Luo Z, Shiojima I, Bialik A, Fulton D, Lefer DJ, Sessa WC, Walsh K. The HMG-CoA reductase inhibitor simvastatin activates the protein kinase Akt and promotes angiogenesis in normocholesterolemic animals. Nat Med 6: 1004–1010, 2000.[CrossRef][Web of Science][Medline]
  17. Laufs U, Fata VL, Plutxky J, Liao JK. Upregulation of endothelial nitric oxide synthase by HMG-CoA reductase inhibitors. Circulation 97: 1129–1135, 1998.[Abstract/Free Full Text]
  18. Masaki I, Yonemitsu Y, Yamashita A, Sata S, Tanii M, Komori K, Nakagawa K, Hou X, Nagai Y, Hasegawa M, Sugimachi K, Sueishi K. Gene therapy for experimental critical limb ischemia: acceleration of limb loss by overexpression of VEGF165 but not of FGF-2. Circ Res 90: 966–973, 2002.[Abstract/Free Full Text]
  19. Mason RP, Walter MF, Jacob RF. Effects of HMG-CoA reductase inhibitors on endothelial function: role of microdomains and oxidative stress. Circulation 109: II34–II41, 2004.[Medline]
  20. Meigs JB, Singer DE, Sullivan LM, Dukes KA, D'Agostino RB, Nathan DM, Wagner EH, Kaplan SH, Greenfield S. Metabolic control and prevalent cardiovascular disease in non-insulin-dependent diabetes mellitus (NIDDM): The NIDDM Patient Outcome Research Team. Am J Med 102: 38–47, 1997.[Web of Science][Medline]
  21. Murohara T, Asahara T, Silver M, Bauters C, Masuda H, Kalka C, Kearney M, Chen D, Symes JF, Fishman MC, Huang PL, Isner JM. Nitric oxide synthase modulates angiogenesis in response to tissue ischemia. J Clin Invest 101: 2567–2578, 1998.[Web of Science][Medline]
  22. Ni W, Egashira K, Kataoka C, Kitamoto S, Koyanagi M, Inoue S, Takeshita A. Antiinflammatory and antiarteriosclerotic actions of HMG-CoA reductase inhibitors in a rat model of chronic inhibition of nitric oxide synthesis. Circ Res 89: 415–421, 2001.[Abstract/Free Full Text]
  23. Onimaru M, Yonemitsu Y, Tanii M, Nakagawa K, Masaki I, Okano S, Ishibashi H, Shirasuna K, Hasegawa M, Sueishi K. FGF-2 gene transfer can stimulate HGF expression, irrespective of hypoxia-mediated down regulation in ischemic limbs. Circ Res 91: 723–730, 2002.
  24. Pitt B, Mancini GB, Ellis SG, Rosman HS, Park JS, McGovern ME. Pravastatin limitation of atherosclerosis in the coronary arteries (PLAC I): reduction in atherosclerosis progression and clinical events. PLAC I investigation. J Am Coll Cardiol 26: 1133–1139, 1995.[Abstract]
  25. Rikitake Y, Liao JK. Rho GTPases, statins, and nitric oxide. Circ Res 97: 1232–1235, 2005.[Abstract/Free Full Text]
  26. Sacks FM, Pfeffer MA, Moye LA, Rouleau JL, Rutherford JD, Cole TG, Brown L, Warnica JW, Arnold JM, Wun CC, Davis BR, Braunwald E. The effect of pravastatin on coronary events after myocardial infarction in patients with average cholesterol levels. Cholesterol and Recurrent Events Trial investigators. N Engl J Med 335: 1001–1009, 1996.[Abstract/Free Full Text]
  27. Sata M, Nishimatsu H, Suzuki E, Sugiura S, Yoshizumi M, Ouchi Y, Hirata Y, Nagai R. Endothelial nitric oxide synthase is essential for the HMG-CoA reductase inhibitor cerivastatin to promote collateral growth in response to ischemia. FASEB J 15: 2530–2532, 2001.[Free Full Text]
  28. Schonbeck U, Libby P. Inflammation, immunity, and HMG-CoA reductase inhibitors: statins as antiinflammatory agents? Circulation 109: II18–II26, 2004.[Medline]
  29. Sparrow CP, Burton CA, Hernandex M, Mundt S, Hassing H, Patel S, Rosa R, Hermanowski-Vosatka A, Wang PR, Zhang D, Peterson L, Detmers PA, Chao YS, Wright SD. Simvastatin has anti-inflammatory and antiatherosclerotic activities independent of plasma cholesterol lowering. Arterioscler Thromb Vasc Biol 21: 115–121, 2001.[Abstract/Free Full Text]
  30. Srinivasan S, Hatley ME, Bolick DT, Palmer LA, Edelstein D, Brownlee M, Hedrick CC. Hyperglycaemia-induced superoxide production decreases eNOS expression via AP-1 activation in aortic endothelial cells. Diabetologia 47: 1727–1734, 2004.[CrossRef][Web of Science][Medline]
  31. Stein E. The lower the better? Reviewing the evidence for more aggressive cholesterol reduction and goal attainment. Atherosclerosis 2: S19–S23, 2002.[CrossRef]
  32. Tanii M, Yonemitsu Y, Fujii T, Shikada Y, Kohne R, Onimaru M, Okano S, Inoue M, Hasegawa M, Onohara T, Maehara Y, Sueishi K. Diabetic microangiopathy in ischemic limb is a disease of disturbance of the platelet-derived growth facto-BB/protein kinase C axis but not of impaired expression of angiogenic factors. Circ Res 98: 55–62, 2006.[Abstract/Free Full Text]
  33. Tsutsumi N, Yonemitsu Y, Shikada Y, Onimaru M, Tanii M, Okano S, Hasegawa M, Maehara Y, Hashizume M, Sueishi K. Essential role of PDGFRa-p70S6K signaling in mesenchymal cells during therapeutic and tumor angiogenesis in vivo: role PDGFRa during angiogenesis. Circ Res 94: 1186–1194, 2004.[Abstract/Free Full Text]
  34. Wolfrum S, Jensen KS, Liao JK. Endothelium-dependent effects of statins. Arterioscler Thromb Vasc Biol 23: 729–736, 2003.[Abstract/Free Full Text]




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