Erythropoietin both protects from and reverses experimental diabetic neuropathy.
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Journal: 2004/February - Proceedings of the National Academy of Sciences of the United States of America
ISSN: 0027-8424
Abstract:
Erythropoietin (EPO) possesses generalized neuroprotective and neurotrophic actions. We tested the efficacy of recombinant human EPO (rhEPO) in preventing and reversing nerve dysfunction in streptozotocin (STZ)-induced diabetes in rats. Two days after STZ [60 mg/kg of body weight (b.w.), i.p.], diabetic animals were administered rhEPO (40 microg/kg of b.w.) three times weekly for 5 weeks either immediately (preventive) before or after a 5-week delay (therapeutic) after induction of hyperglycemia or at a lower dose (8 microg/kg of b.w. once per week) for 8 weeks (prolonged). Tail-nerve conduction velocities (NCV) was assessed at 5 and 11 weeks for the preventive and therapeutic schedule, respectively. Compared to nondiabetic rats, NCV was 20% lower after 5 weeks in the STZ group, and this decrease was attenuated 50% by rhEPO. Furthermore, the reduction of Na(+),K(+)-ATPase activity of diabetic nerves (by 55%) was limited to 24% in the rhEPO-treated group. In the therapeutic schedule, NCV was reduced by 50% after 11 weeks but by only 23% in the rhEPO-treated group. rhEPO treatment attenuated the decrease in compound muscle action potential in diabetic rats. In addition, rhEPO treatment was associated with a preservation of footpad cutaneous innervation, as assessed by protein gene product 9.5 immunostaining. Diabetic rats developed alterations in mechanical and thermal nociception, which were partially reversed by rhEPO given either in a preventative or therapeutic manner. These observations suggest that administration of rhEPO or its analogues may be useful in the treatment of diabetic neuropathy.
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Proc Natl Acad Sci U S A 101(3): 823-828
PMID: 14718663

Erythropoietin both protects from and reverses experimental diabetic neuropathy

+6 authors
Mario Negri Institute of Pharmacological Research, 20157 Milan, Italy; Departments of Biophysics, Biochemistry, Biostatistics, and Neurology, Mersin University, 33079 Mersin, Turkey; Kenneth S. Warren Institute, Kitchawan, NY 10653; University of Milan “Bicocca,” 20052 Monza, Italy; National Neurological Institute, 20133 Milan, Italy; and University of Brescia, 25125 Brescia, Italy
To whom correspondence should be addressed. E-mail: ti.irgenoiram@aibbor.
Present address: Novuspharma S.p.A., 20091 Bresso, Italy.
Contributed by Anthony Cerami, November 25, 2003
Contributed by Anthony Cerami, November 25, 2003
Copyright © 2004, The National Academy of Sciences

Abstract

Erythropoietin (EPO) possesses generalized neuroprotective and neurotrophic actions. We tested the efficacy of recombinant human EPO (rhEPO) in preventing and reversing nerve dysfunction in streptozotocin (STZ)-induced diabetes in rats. Two days after STZ [60 mg/kg of body weight (b.w.), i.p.], diabetic animals were administered rhEPO (40 μg/kg of b.w.) three times weekly for 5 weeks either immediately (preventive) before or after a 5-week delay (therapeutic) after induction of hyperglycemia or at a lower dose (8 μg/kg of b.w. once per week) for 8 weeks (prolonged). Tail-nerve conduction velocities (NCV) was assessed at 5 and 11 weeks for the preventive and therapeutic schedule, respectively. Compared to nondiabetic rats, NCV was 20% lower after 5 weeks in the STZ group, and this decrease was attenuated 50% by rhEPO. Furthermore, the reduction of Na,K-ATPase activity of diabetic nerves (by 55%) was limited to 24% in the rhEPO-treated group. In the therapeutic schedule, NCV was reduced by 50% after 11 weeks but by only 23% in the rhEPO-treated group. rhEPO treatment attenuated the decrease in compound muscle action potential in diabetic rats. In addition, rhEPO treatment was associated with a preservation of footpad cutaneous innervation, as assessed by protein gene product 9.5 immunostaining. Diabetic rats developed alterations in mechanical and thermal nociception, which were partially reversed by rhEPO given either in a preventative or therapeutic manner. These observations suggest that administration of rhEPO or its analogues may be useful in the treatment of diabetic neuropathy.

Abstract

Polyneuropathy is the most common complication of diabetes mellitus, occurring in >50% of patients who have been hyperglycemic for >15 years (1, 2). Neuropathy contributes the greatest morbidity and mortality and severely impairs the quality of life (3, 4) because of paresthesia, pain, and neuropathic injury, the leading cause of nontraumatic amputation in the U.S. Hyperglycemia is critical for the development and progression of diabetic neuropathy (1, 2), with the two main pathogenic hypotheses focusing on a metabolic vs. vascular etiology. Despite many studies of human and experimental diabetic neuropathy, the current therapeutic arsenal is very poor.

We have previously shown that recombinant human erythropoietin (rhEPO) crosses the blood–brain barrier and has a protective effect in animal models of cerebral ischemia and traumatic injury (5). In primary neuronal cultures or neuronal cell lines and in cerebral ischemia, rhEPO protects from apoptosis (6, 7). rhEPO also reduces injury in experimental autoimmune encephalomyelitis, injury of spinal cord, or sciatic nerve compression (8). i.v. rhEPO is well tolerated and beneficial in patients with acute ischemic stroke (9). Prior work has documented that, within the sciatic nerve, both neurons and Schwann cells express the EPO receptor, which is up-regulated after injury (10). Furthermore, a potential beneficial role for rhEPO in mechanical peripheral nerve injury has been recently shown, for which rhEPO treatment protects dorsal root ganglion neurons from undergoing apoptosis (11).

The present study is aimed at investigating the efficacy of rhEPO in preventing and/or treating peripheral diabetic neuropathy. To accomplish this, we studied rats with streptozotocin (STZ)-induced diabetes, which, like human diabetic neuropathy, have alterations in nociceptive thresholds (thermal and mechanical). In this model, we evaluated thermal and mechanical nociceptive thresholds, supplemented by nerve conduction velocity (NCV) and compound muscle action potentials (CMAP), and sciatic nerve Na,K-ATPase content.

Some authorities have considered the first fibers affected in diabetic neuropathy to be the small sensory ones (12–14). In addition, recent data show that the degeneration of intraepidermal nerve fiber (IENF), somatic unmyelinated axons, correlates with both the presence and severity of sensory neuropathy (15–18). We have assessed the involvement of IENF in the STZ-diabetic rat model by quantifying density in rat footpad skin by use of protein gene product 9.5 (PGP 9.5), a specific marker. Administration of rhEPO in all these experiments reduces loss of function in the diabetic state.

Acknowledgments

We thank Dr. N. A. Calcutt for helpful suggestions, especially on measuring nociceptive thresholds, and Mehmet Acioglu of the Research Laboratory at Mersin for assistance. This work was supported in part by Ministero dell'Istruzione, dell'Università e della Ricerca, Rome (Fondo Integrativo Speciale per la Ricerca and Fondo per gli Investimenti della Ricerca di Base and RBAU01AR5J) and by Fondo Integrativo Speciale per la Ricerca–Neurobiotecnologie from the Ministero dell'Istruzione, dell'Università e della Ricerca. The work was also supported by a grant from the Kenneth S. Warren Institute.

Acknowledgments

Notes

Abbreviations: STZ, streptozotocin; EPO, erythropoietin; rhEPO, recombinant human EPO; CMAP, compound muscle action potential; IENF, intraepidermal nerve fiber; NCV, nerve conduction velocity; PGP 9.5, protein gene product 9.5; b.w., body weight.

Notes
Abbreviations: STZ, streptozotocin; EPO, erythropoietin; rhEPO, recombinant human EPO; CMAP, compound muscle action potential; IENF, intraepidermal nerve fiber; NCV, nerve conduction velocity; PGP 9.5, protein gene product 9.5; b.w., body weight.

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