Vasoconstriction, RhoA/Rho-kinase and the erectile response

Mills, Thomas M.; Lewis, Ronald W.; Wingard, Christopher J.; Linder, Anna; Jin, Liming; Webb, R. Clinton

doi:10.1038/sj.ijir.3901068

Cited by 31 publications

(32 citation statements)

References 30 publications

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“…ROCK inhibitors, like Y-27632 and fasudil, 15) can selectively bind to the catalytic domain and inhibit its activity. ROCK inhibitors were regarded as promising choice to treat nerve injury, 16) myocardial injury, 8) erectile dysfunction, 7) and other diseases. Therefore, purified ROCK-CD, as an executive domain, is very useful in screening ROCK inhibitors and studying ROCK function in vitro.…”

Section: Discussionmentioning

confidence: 99%

“…4) Rho kinase is thought to play an important role in a variety of cellar functions such as stress fiber formation, focal adhesion formation, cell aggregation, cell morphology, cytokinesis, cell migration, and Ca 2ϩ -sensitization in the smooth muscle. 2,5) Recent report showed that Rho kinase involves in regulation of the amyloid precursor protein processing.6) ROCK inhibitors are beneficial candidates for a wide range of diseases such as hypertension, inflammation, cancer, 4) erectile dysfunction, 7) and injury caused by ischemia and reperfusion. …”

mentioning

confidence: 99%

“…6) ROCK inhibitors are beneficial candidates for a wide range of diseases such as hypertension, inflammation, cancer, 4) erectile dysfunction, 7) and injury caused by ischemia and reperfusion.…”

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confidence: 99%

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Harvest Active Recombinant Rho Kinase from Escherichia coli

Duan

Wang

Chen

et al. 2006

Biological & Pharmaceutical Bulletin

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Rho kinase, known as Rho-associated coiled-coil forming protein serine/threonine kinase (ROCK), 1) is one of the central regulatory molecules for cytoskeleton control and cell adhesion process.2) ROCK also involves in other cell functions such as apoptosis 3) and tumor invasiveness. 4) Rho kinase is thought to play an important role in a variety of cellar functions such as stress fiber formation, focal adhesion formation, cell aggregation, cell morphology, cytokinesis, cell migration, and Ca 2ϩ -sensitization in the smooth muscle. 2,5) Recent report showed that Rho kinase involves in regulation of the amyloid precursor protein processing.6) ROCK inhibitors are beneficial candidates for a wide range of diseases such as hypertension, inflammation, cancer, 4) erectile dysfunction, 7) and injury caused by ischemia and reperfusion. 8)There are two types of ROCK: ROCK-I (ROCK-b) mainly found in hepatocytes, and ROCK-II (ROCK-a) in central nerve system (CNS). ROCK is activated by binding with the activated form of a low molecular weight G protein, Rho. 9)Purified ROCK is important foundation for understanding the full function of ROCK, and is very useful in ROCK inhibitors screening. Katoh et al. 10) have conformed that 1-543 amino acid residua have catalytic function; further research suggested the kernel catalytic domain of ROCK (ROCK-CD) may be 92-354 amino acid residua.11) Turner et al. 12) reported a way to harvest recombinant human ROCK-CD from Sf-21 cell, which became a main purified ROCK source currently. However, this procedure is timeconsuming and at a relatively high cost; which makes its price relatively expensive. It is needed to improve a procedure to harvest the ROCK-CD at low price, at high quality, and at large quantity; harvesting active ROCK-CD from Escherichia coli (E. coli) may be the choice met the request. MATERIALS AND METHODS RT-PCRThe cDNA encoding the amino acids 5-552 of rat ROCK-II was cloned with polymerase chain reaction (PCR) techniques. Primers were designed according to rat ROCK-II sequence NM_009072 in GenBank.11) Briefly, Rat brain total RNA was extracted by TRIzol kit (Invitrogen, U.S.A.), cDNA was reverse transcribed by reverse transcriptase from rat brain total RNA. A partial cDNA fragment of rat ROCK-II, 1644 nt (13-1656) was amplified using Taq DNA Polymerase (Takara Biotech, Japan) with 10 pM of each primer (cycling conditions: 94°C for 60 s, 55°C for 60 s, 72°C for 90 s, 30 cycles, finally kept at 72°C for 20 min). Primer A was 5Ј-ATGAGCGGATCCCCGCCGACGGGGA-AAAT-3Ј and primer B was 5Ј-ACCTCTCTCGAGTATCT-GAGAGCTCTGGT-3Ј (the underline in primer A is BamHI site; and in primer B is XholI site). The PCR-amplified fragment was subcloned into pGEM-T Easy Vector (Promega, U.S.A.). Then, the cDNA of ROCK-CD was sequenced by the vector and confirmed as the encoding 5-552 amino acids.Positive Clone Screening ROCK-CD cDNA in pGEM-T Easy Vector was digested with BamHI and XhoI and ligated into pET28a(ϩ) expression vector (Novagen, U.S.A.). The pET28a(ϩ) contains a (His)6-tag at its N-t...

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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Harvest Active Recombinant Rho Kinase from Escherichia coli

Duan

Wang

Chen

et al. 2006

Biological & Pharmaceutical Bulletin

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“…In this way, RhoA/Rho-kinase activation has significant effects on various cardiovascular diseases, such as arterial hypertension (133) (140) and ED. Since the main function of Rhokinase is the regulation of smooth muscle tone (141), the upregulation of the Rho-kinase pathway increases cavernosal smooth muscle contraction, leading to ED (142)(143). Furthermore, studies indicate that Rho-kinase isoforms are activated in patients with a cardiovascular disorder or associated risk factors (144)(145).…”

Section: The Link Between Ed and Cvdmentioning

confidence: 99%

The Role Erectile Dysfunction Plays in Cardiovascular Diseases

Crestani¹,

Nunes²,

Marques³

et al. 2012

Erectile Dysfunction - Disease-Associated Mechanisms and Novel Insights Into Therapy

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“…2,4 Smooth muscle contraction is mediated by phosphorylation of myosin light chain (MLC) via the Ca 2 þ /calmodulin-dependent activation of MLC kinase and actin/myosin cross-bridge formation. 51,52 Smooth muscle relaxation is mediated by dephosphorylation of MLC via MLC phosphatase. 53 A principle regulator of MLC phosphatase is Rho-kinase, which belongs to the serine/threonine kinase family.…”

Section: Sod Gene Therapy For Edmentioning

confidence: 99%

Gene and stem cell therapy for erectile dysfunction

2005

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Erectile dysfunction (ED) is defined as the inability to attain and/or maintain penile erection sufficient for satisfactory sexual performance. ED is a highly prevalent health problem with considerable impact on the quality of life of men and their partners. Although the treatment of ED with oral phosphodiesterase type V (PDE5) inhibitors is effective in a wide range of individuals, it is not efficacious in all patients. The failure of PDE5 inhibitors happens mainly in men with diabetes, non-nerve sparing radical prostatectomy, and high disease severity. Therefore, improved therapies based on a better understanding of the fundamental issues in erectile physiology and pathophysiology have recently been proposed. Here, we summarize studies on ED treatment using gene and stem cell therapies. Adenoviral-mediated intracavernosal transfer of therapeutic genes, such as endothelial nitric oxide synthase (eNOS), calcitonin gene-related peptide (CGRP), superoxide dismutase (SOD), and RhoA/Rho kinase and mesenchymal stem cell-based cell and gene therapy strategy for the treatment of age-and diabetes-related ED are the focus of this review. Penile erection, referred to as engorgement of the penis with blood, is the result of the penile arterial dilation which increases blood inflow, the relaxation of smooth muscle of the corpus cavernosum which increases blood storage, and penile venous occlusion which decreases blood outflow. The physiology of penile erection is complicated and this neurovascular event requires the functional integrity of nerves, endothelium, and smooth muscle. 1 Nitric oxide (NO), produced by both neuronal nitric oxide synthase (nNOS) in the nonadrenergic noncholinergic (NANC) nerves and endothelial nitric oxide synthase (eNOS) in the endothelium of penile arteries and cavernosal sinusoids is the principal mediator of penile erection. 2-5 NO formed in nerves or endothelial cells then diffuses to neighboring smooth muscle cells and activates soluble guanylyl cyclase. This activation results in increased cGMP formation, which mediates smooth muscle relaxation. 5 Besides NO, other molecules such as vasoactive intestinal polypeptide (VIP), calcitonin gene-related peptide (CGRP), substance P, and pituitary adenylate cyclase-activating polypeptide (PACAP) are also important for penile erection. 6 Causes contributing to erectile dysfunction (ED) can be broadly classified into two categories: organic and psychological. Aging, vascular diseases, neurological injuries, and diabetes mellitus are the common causes of organic ED. 7 Depressive stress is the common cause of psychological ED. 8,9 Although the treatment of ED with oral phosphodiesterase type V (PDE5) inhibitors (sildenafil, tadalafil, and vardenafil) is effective in a wide range of individuals with ED, it is not efficacious in all patients. For example, the response rate to sildenafil decreased from 72% in men 18-49 y of age to 53% in men 50 y or older. 10 The failure of sildenafil therapy happened mainly in men with diabetes, non-nerve sparing radical pr...

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Vasoconstriction, RhoA/Rho-kinase and the erectile response

Cited by 31 publications

References 30 publications

Harvest Active Recombinant Rho Kinase from Escherichia coli

Harvest Active Recombinant Rho Kinase from Escherichia coli

The Role Erectile Dysfunction Plays in Cardiovascular Diseases

Gene and stem cell therapy for erectile dysfunction

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