Bile acids regulate anorexigenic neuropeptide through p-STAT3-SOCS3 signaling in mouse hypothalamic cells / 浙江大学学报·医学版

OBJECTIVE@#To explore the effects of taurolithocholic acid (tLCA) and chenodeoxycholic acid (CDCA) on the expression of aorexigenic neuropeptide in mouse hypothalamus GT1-7 cells.@*METHODS@#Mouse hypothalamic GT1-7 cells were treated with culture medium containing 10% FBS (control group, =3) or with 10 nmol/L, 100 nmol/L, 1 μmol/L and 10 μmol/L tLCA (tLCA group, =3) or CDCA (CDCA group, =3) for 12, 24 or 48 h. Real-time PCR was performed to determine the expression levels of proopiomelanocortin (POMC) mRNA in the cells, and the production levels of α-melanocyte-stimulating hormone (α-MSH) were assessed using an ELISA kit. Signal transduction and activator of transcription 3 phosphorylation (p-STAT3), threonine kinase phosphorylation (p-AKT), suppressor of cytokine signaling 3 (SOCS3), G protein-coupled bile acid receptor-1 (TGR5) and farnesoid X receptor (FXR) protein were detected by Western blotting.@*RESULTS@#Western blotting results showed that mouse hypothalamic GT1-7 cells expressed two bile acid receptors, TGR5 and FXR, whose expressions were regulated by bile acids. Real-time PCR showed that the expression of POMC mRNA was significantly increased in the cells after treatment with 10 μmol/L tLCA or CDCA for 24 h. POMC-derived anorexigenic peptide α-MSH increased significantly in GT1-7 cells after treatment with 10 μmol/L tLCA or CDCA for 24 h. Treatment of the cells with tLCA or CDCA significantly increased the expressions of intracellular signaling proteins including p-STAT3, p-AKT and SOCS3.@*CONCLUSIONS@#Mouse hypothalamic GT1-7 cells express bile acid receptors TGR5 and FXR. Bile acids tLCA or CDCA can promote the expression of POMC mRNA and increase the production of the anorexigenic peptide α-MSH. The intracellular signaling proteins p-AKT, p-STAT3 and SOCS3 are likely involved in bile acid-induced anorexigenic peptide production.

Participação do hormônio liberador de corticotropina (CRH) e dos hormônios da pró-opiomelanocortina (POMC) no lúpus eritematoso sistêmico com envolvimento cutâneo / CRH and pro-opiomelanocortin (POMC) participation in systemic lupus erythematosus with skin involvement

Introdução: A ativação do eixo hormônio liberador de corticotropina (CRH) e da pró-opiomelanocortina (POMC) leva a produção de vários derivados bioativos que incluem o hormônio adrenocorticotrófico (ACTH) e o hormônio estimulador de melanócito alfa (alfa-MSH). Estudos avaliando a participação desse eixo no lúpus eritematoso sistêmico (LES) são escassos, particularmente no envolvimento cutâneo da doença. Objetivo: Avaliar a participação do CRH e das melanocortinas (MCs) na fisiopatologia do lúpus eritematoso sistêmico com envolvimento cutâneo. Métodos: Dezessete pacientes com LES com envolvimento cutâneo foram avaliados clinicamente e biópsias da pele afetada e não afetada e do sangue periférico foram obtidas. Dezessete indivíduos saudáveis foram pareados por idade e gênero. Os fragmentos de pele foram submetidos à análise imuno-histoquímica para avaliação da expressão de CRH, ACTH, alfaMSH, e receptor de melanocortina tipo 1 (MC-1R). Os níveis séricos de alfa-MSH, IL-1, IL-1ra, IL-6, IL-10, IL-12p70, IL-17, TNF-alfa, e IFN-y foram determinados pelo método Multiplex. Resultados: A pele afetada de pacientes com LES apresentaram maior expressão CRH na derme profunda quando comparada à pele não afetada dos mesmos doentes e a pele saudável dos controles (p = 0,024). Níveis séricos de alfa-MSH foram similares entre LES e controles. Dentre as citocinas avaliadas, IFN-y, TNF-alfa e IL-6 foram mais elevadas nos pacientes com LES em relação aos controles (p = 0,041, p = 0,001 e p = 0,049, respectivamente). Embora não significativamente, os níveis de IL-17 também foram mais altos nos pacientes (p = 0,099). A expressão tecidual de ACTH, cortisol, alfa-MSH e seu receptor MC-1R foram semelhantes entre os pacientes e controles. Conclusões: Nossos resultados mostram, pela primeira vez a participação do eixo CRH-POMC na patogênese das lesões cutâneas do LES.

Introduction: Corticotropin-releasing hormone (CRH) and pro-opiomelanocortin (POMC) axis activation leads to the production of several bioactive hormones including adrenocorticotrophic hormone (ACTH) and the neuropeptide alfa-melanocyte stimulating hormone (alfa-MSH). There are scarce data regarding their role in systemic lupus erythematosus (SLE) particularly in cutaneous involvement of this disease. Objective: To evaluate the role of CRH and melanocortins (MCs) in the pathophysiology of systemic lupus erythematosus with skin involvement. Methods: Seventeen patients with SLE with skin involvement were evaluated clinically and biopsies of affected and unaffected skin and peripheral blood were obtained. Seventeen healthy subjects were matched for age and gender. The skin fragments were subjected to immunohistochemical analysis for the expression of CRH, ACTH, alfa-MSH and melanocortin receptor type 1 (MC-1R). Serum levels of alfa-MSH, IL-1, IL-1ra, IL-6, IL-10, IL-12p70, IL-17, TNF-alfa and IFN-y were determined by multiplex. Results: The affected skin of SLE patients exhibited greater CRH expression in the deep dermis compared to unaffected skin of the same patients and the control's healthy skin (p = 0.024). alfa-MSH were similar between SLE and controls. Among the evaluated cytokines, IFN-y, TNF-alfa and IL-6 were significantly higher in SLE patients compared to controls (p = 0.041, p = 0.001 and p = 0.049, respectively). Although not significant, levels of IL-17 were also higher in patients (p = 0.099). Tissue expression of ACTH, cortisol, alfa-MSH and its receptor MC-1R were similar between patients and controls. Conclusions: Our results show for the first time the involvement of CRH-POMC axis in the pathogenesis of SLE cutaneous lesions through interactions between the brain-skin axis.

Superoxide Dismutase 1 Inhibits Alpha-Melanocyte Stimulating Hormone and Ultraviolet B-Induced Melanogenesis in Murine Skin

BACKGROUND: Over the last decade, the incidence of ultraviolet B (UVB)-related skin problems has increased. Oxidative stress caused by UVB induces the secretion of melanocyte growth and activating factors from keratinocytes, which results in the formation of cutaneous hyperpigmentation. Therefore, increasing the antioxidant abilities of skin cells is thought to be a beneficial strategy for the development of sunscreen agents. Superoxide dismutase 1 (SOD1) is an antioxidant enzyme that is known to exhibit antioxidant properties. OBJECTIVE: The purpose of this study was to investigate the effect of SOD1 on alpha-melanocyte stimulating hormone (alpha-MSH) and UVB-induced melanogenesis in B16F10 melanoma cells and HRM-2 melanin-possessing hairless mice. METHODS: The inhibitory effect of SOD1 on tyrosinase activity was evaluated in a cell-free system. Additional experiments were performed using B16F10 melanoma cells to demonstrate the effects of SOD1 in vitro, and HRM-2 melanin-possessing hairless mice were used to evaluate the antimelanogenic effects of SOD1 in vivo. RESULTS: We found that SOD1 inhibited melanin production in a dose-dependent manner without causing cytotoxicity in B16F10 melanoma cells. SOD1 did not inhibit tyrosinase activity under cell-free conditions. The results indicate that SOD1 may reduce pigmentation by an indirect, nonenzymatic mechanism. We also found that SOD1 decreased UVB-induced melanogenesis in HRM-2 melanin-possessing hairless mice, as visualized through hematoxylin and eosin staining and Fontana-Masson staining. CONCLUSION: Our results indicate that SOD1 has an inhibitory effect on alpha-MSH and UVB-induced melanogenesis, indicating that SOD1 may be a promising sunscreen agent.

Development of intestinal ischemia/reperfusion-induced acute kidney injury in rats with or without chronic kidney disease: Cytokine/chemokine response and effect of alpha-melanocyte-stimulating hormone

BACKGROUND: The primary aim of the study was to investigate the cytokine/chemokine response in the kidney, lung, and liver following acute kidney injury (AKI). The secondary aim was to test whether alpha-melanocyte-stimulating hormone (alpha-MSH) could prevent a reduction in organ function, and attenuate the inflammatory cytokine/chemokine response within the kidney, lung, and liver following AKI in rats with or without preexisting chronic kidney disease (CKD). METHODS: A two-stage animal model, in which AKI was induced in rats with preexisting CKD, induced by 5/6 nephrectomy (Nx), was used. Six weeks later, AKI was induced by intestinal ischemia and reperfusion (IIR). Sham procedures [S(Nx) and S(IIR)] were also performed. RESULTS: Increasing levels of serum creatinine (sCr) demonstrated progressive development of CKD in response to Nx, and following IIR sCr levels increased further significantly, except in the S(Nx) group treated with alpha-MSH. However, no significant differences in the fractional increase in sCr were observed between any of the groups exposed to IIR. In kidney, lung, and liver tissue the levels of interleukin (IL)-1beta were significantly higher in rats undergoing IIR when compared to the S(IIR) and control rats. The same pattern was observed for the chemokine monocyte chemoattractant protein (MCP)-1 in lung and liver tissue. Furthermore, kidney IL-1beta and RANTES levels were significantly increased after IIR in the Nx rats compared to the S(Nx) rats. CONCLUSION: Both the functional parameters and the cytokine/chemokine response are as dramatic when AKI is superimposed onto CKD as onto non-CKD. No convincing protective effect of alpha-MSH was detected.

Inhibitory Effects of Resveratrol on Melanin Synthesis in Ultraviolet B-Induced Pigmentation in Guinea Pig Skin

Resveratrol is a polyphenolic compound found in various natural products such as grapes and berries and possesses anti-cancer, anti-hyperlipidemia, and anti-aging properties. Recently, it has been reported that resveratrol inhibits alpha-melanocyte-stimulating hormone signaling, viability, and migration in melanoma cells. However, these effects have not been confirmed in vivo, specifically brownish guinea pigs. To evaluate the potential of resveratrol as a regulator of melanin for hyperpigmentation therapy, the influence of resveratrol on pigmentation was investigated by ultraviolet B-induced hyperpigmentation in brownish guinea pig skin. We found that resveratrol reduced the expression of melanogenesis-related proteins tyrosinase, tyrosinase-related proteins 1 and 2, and microphthalmia-associated transcription factor in melanoma cells. Furthermore, topical application of resveratrol was demonstrated to significantly decrease hyperpigmentation on ultraviolet B-stimulated guinea pig skin in vivo. Based on our histological data, resveratrol inhibits melanin synthesis via a reduction in tyrosinase-related protein 2 among the melanogenic enzymes. This study is the first to provide evidence supporting resveratrol as a depigmentation agent, along with further clinical investigation of resveratrol in ultraviolet B-induced skin disorders such as hyperpigmentation and skin photoaging.

Inhibitory Effects of Resveratrol on Melanin Synthesis in Ultraviolet B-Induced Pigmentation in Guinea Pig Skin

Resveratrol is a polyphenolic compound found in various natural products such as grapes and berries and possesses anti-cancer, anti-hyperlipidemia, and anti-aging properties. Recently, it has been reported that resveratrol inhibits alpha-melanocyte-stimulating hormone signaling, viability, and migration in melanoma cells. However, these effects have not been confirmed in vivo, specifically brownish guinea pigs. To evaluate the potential of resveratrol as a regulator of melanin for hyperpigmentation therapy, the influence of resveratrol on pigmentation was investigated by ultraviolet B-induced hyperpigmentation in brownish guinea pig skin. We found that resveratrol reduced the expression of melanogenesis-related proteins tyrosinase, tyrosinase-related proteins 1 and 2, and microphthalmia-associated transcription factor in melanoma cells. Furthermore, topical application of resveratrol was demonstrated to significantly decrease hyperpigmentation on ultraviolet B-stimulated guinea pig skin in vivo. Based on our histological data, resveratrol inhibits melanin synthesis via a reduction in tyrosinase-related protein 2 among the melanogenic enzymes. This study is the first to provide evidence supporting resveratrol as a depigmentation agent, along with further clinical investigation of resveratrol in ultraviolet B-induced skin disorders such as hyperpigmentation and skin photoaging.

Induction of Melanogenesis by Rapamycin in Human MNT-1 Melanoma Cells

BACKGROUND: Melanogenesis is one of the characteristic parameters of differentiation in melanocytes and melanoma cells. Specific inhibitors of phosphatidylinositol 3-kinase (PI3K), such as wortmannin and LY294002, stimulate melanin production in mouse and in human melanoma cells, suggesting that PI3K and mammalian target of rapamycin (mTOR) might be involved in the regulation of melanogenesis. OBJECTIVE: The involvement of the mTOR pathway in regulating melanogenesis was examined using human MNT-1 melanoma cells, and the effects of the potent inhibitor of mTOR, rapamycin, in the presence or absence of alpha-melanocyte-stimulating hormone (alpha-MSH) were evaluated. METHODS: In cells treated with rapamycin, cell viability, melanin content, and tyrosinase (TYR) activity were measured and compared with untreated controls. Protein levels of TYR, tyrosinase-related protein (TYRP)-1, TYRP-2, and microphthalmia-associated transcription factor (MITF) were also analyzed by Western blot. RESULTS: In rapamycin-treated cells, the melanin content increased concomitantly with an elevation in TYR activity, which plays a major role in melanogenesis. There was also an up-regulation of TYR, TYRP-1, and MITF proteins. Combined treatment with rapamycin or wortmannin and alpha-MSH increased melanogenesis more strongly than alpha-MSH alone. CONCLUSION: Rapamycin-induced melanin formation may be mediated through the up-regulation of TYR protein and activity. Furthermore, rapamycin and wortmannin, inhibitors of mTOR and PI3K, respectively, have co-stimulatory effects with alpha-MSH in enhancing melanogenesis in melanocyte cells.

Effect of xanthohumol on melanogenesis in B16 melanoma cells

Xanthohumol (XH), the principal prenylflavonoid of the hop plant (Humulus lupulus L.), dose-dependently inhibited isobutylmethylxanthine (IBMX)-induced melanogenesis in B16 melanoma cells, with little cytotoxicity at the effective concentrations. Decreased melanin content was accompanied by reduced tyrosinase enzyme activity, protein and mRNA expression. The levels of tyrosinase-related protein 1 and 2 mRNAs were decreased by XH. XH also inhibited alpha-melanocyte stimulating hormone- or forskolin-induced increases in melanogenesis, suggesting an action on the cAMP-dependent melanogenic pathway. XH downregulated the protein and mRNA expression of microphthalmia-associated transcription factor (MITF), a master transcriptional regulator of key melanogenic enzymes. These results suggest that XH might act as a hypo-pigmenting agent through the downregulation of MITF in the cAMP-dependent melanogenic pathway.

Detection of binding activity and biologic effect of a novel alpha-melanocyte-stimulating hormone analogue / 药学学报

Binding activity and biologic effect of a novel alpha-melanocyte-stimulating hormone analogue were tested on cells transiently expressing the human melanocortin-1 (MC1), MC3, MC4, and MC5 receptors. The human MC1 and MC5 receptor genes were cloned into the expression vector pcDNA3. 1/ myc-his(-) B. The vectors were transferred to HEK-293 cells by the calcium phosphate method. Stable receptor populations were generated using G418 selection (900 microg x mL(-1)) for subsequent bioassay analysis. K(i) values of the novel alpha-MSH analogue for MC1, MC3, MC4, and MC5 receptors were obtained in competition with [125I]-NDP-MSH for binding studies. The cyclic AMP level was tested by using [3H]-cyclic AMP kit. It is showed that K(i) values of the novel alpha-MSH analogue for MC1, MC3, MC4, and MC5 receptors were (0.159 +/- 0.040), (35.430 +/- 6.743), (19.293 +/- 2.780) and (2.230 +/- 0.670) nmol L(-1), respectively. Its EC50 values for MC1, MC3, MC4, and MC5 receptors were (0.45 +/- 0.07), (7.80 +/- 0.65), (2.55 +/- 0.23) and (0.33 +/- 0.09) nmol L(-1), respectively. In these tests, the novel alpha-MSH analogue is a MC1R and MC5R selective agonist.

Effects of bFGF and alpha-MSH on adhesion and migration of human melanocytes in vitro / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To observe the effect of basic fibroblast growth factor (bFGF) and alpha-melanocyte stimulating hormone (alpha-MSH) on adhesion and migration of melanocytes in vitro.</p><p><b>METHODS</b>Human melanocytes were obtained from normal human foreskins. Culture dishes covered with fibronectin were used to perform melanocytes adhesion assay, and cell motility was assessed using the Transwell micropore filter method.</p><p><b>RESULT</b>bFGF and alpha-MSH increased melanocytes adhesion on culture dishes covered with fibronectin. bFGF stimulated melanocytes migration through micropore filter while alpha-MSH had no significant effects.</p><p><b>CONCLUSION</b>bFGF and alpha-MSH could promote the adhesion and migration of melanocytes, which suggests that two agents may play a role in the repigmentation of vitiligo.</p>

Alpha-Melanocyte Stimulating Hormone Protects Pancreatic Islet Dysfunction by Peripheral Blood Mononuclear Cells in vitro / 대한이식학회지

PURPOSE: The alpha-melanocyte-stimulating hormone (alpha-MSH) has been shown to interact with various cells of the immune and inflammatory system and down-regulate either the production or the action of the pro-inflammatory cytokines. In this study, we investigated the potential of alpha-MSH on preventing pancreatic islet cell from death and dysfunction by inflammatory cytokines released from peripheral blood mononuclear cells (PBMCs) in rat. METHODS: Rat pancreatic islets were co-cultured with PBMCs, stimulated by phorbol myrstic acid and ionomycin. alpha-MSH was treated to PBMCs for 2 hours before co-culture. Viability and apoptosis of islets were observed by MTT and FACS. Inflammatory cytokines and nitric oxide (NO) were measured. Insulin release from islet co-cultured with mononuclear cells was checked for the islet function. RESULTS: In comparison to control group, viability of islets with alpha-MSH treated mononuclear cells was increased and apoptosis was reduced significantly. Inflammatory cytokines such as TNF-alpha and IL-1beta were reduced in alpha-MSH-treated group. NO production in alpha-MSH-treated group was decreased. Insulin secretory function of islet was recovered in condition of alpha-MSH treatment. CONCLUSION: This study demonstrates that alpha-MSH protects cell death and preserves the secretory function of pancreatic islet cells from the pro-inflammatory reaction of mononuclear cells, and may have the potential to improve the graft survival in clinical islet transplantation.

Expression of Human Melanocyte Markers in PMA-containing Medium and Physiologic Mitogen-containing Medium / 대한피부과학회지

BACKGROUND: There are many growth media for cultivation of human melanocytes (MGM), depending on the supplements added, and the growth of cells is closely related to these components. To understand melanocytes in vivo, it is necessary to find out the biological or biochemical characteristics of melanocytes grown in physiologic growth medium (P-MGM) and phorbol-12-myristate 13-acetate (PMA)-containing medium (C-MGM). OBJECTIVE: To investigate the expression of different biochemical markers of melanocytes grown in C-MGM and in P-MGM. METHODS: C-MGM is basically composed of PMA (10 ng/ml), and bFGF (3 ng/ml), and is now commercially available for melanocyte culture. P-MGM is a physiologic growth medium containing physiologic mitogens such as bFGF (10 ng/ml), ET-1 (10 nM), and alpha-MSH (12 nM). The cell proliferation and the expression of biochemical markers were measured in cultured human melanocytes which were grown in either C-MGM or P-MGM. RESULTS: In this study, there was significant difference in cell proliferation between cells grown in C-MGM and P-MGM (p<0.01). The tyrosinase activity and melanin contents were significantly increased in C-MGM. The expression of TRP1, MART-1 and p53 in mRNA level was higher in C-MGM than in P-MGM. The up-regulation of p53 protein expression was also observed in C-MGM. CONCLUSION: The proliferation and expression of p53, at both transcriptional and translational levels were increased when melanocytes were grown in C-MGM, compared to P-MGM. This data suggests that p53-mediated melanization is to some degree related with phorbol ester, and should further be elucidated.

The Effect of alpha-Melanocyte Stimulating Hormone on Renal Ischemia Reperfusion Injury / 대한이식학회지

PURPOSE: In ischemia-reperfusion induced renal injuries, cytokines, chemoattractant chemokines, adhesion molecules and nitric oxide play an important role. alpha-Melanocyte stimulating hormone (alpha-MSH) is a potent anti-inflammatory cytokine so the therapeutic effect of alpha-MSH on an ischemia-reperfusion induced acute renal failure is to be evaluated. METHODS: 40 male Spraque-Dawley rats were prepared for the experiment, they were classified into three classes (Sham, ischemic control and alpha-MSH injection). Both renal pedicles were clamped for 45 minutes. alpha-MSH (50 microgram) was injected intravenously three times, immediately before ischemia and reperfusion and 18 hour after reperfusion. Serum creatinine and histologic changes were analyzed between groups (Sham (n=6), ischemic control group (n=15), and alpha-MSH group (n=19)). RESULTS: Serum creatinine level decreased significantly at 24 hours after reperfusion in alpha-MSH treated animals (SCr24 0.78+/-0.23 mg/dL, 4.21+/-1.14 mg/dL, 3.01+/-1.19 mg/dL, repectively (P=0.008)), especially serum creatinine level at 48 hours after reperfusion much more dicreased in alpha-MSH group (SCr48 0.67+/-0.16 mg/dL, 4.21+/-2.03 mg/dL, 1.15+/-1.11 mg/dL, repectively (P=0.004)). Tubular neccrosis and neutrophil infiltration decreased signigicantly in alpha-MSH treated group (P=0.001). Mortality was noted 33.3% only at ischemic conrol group. CONCLUSION: we demonstrate the fact that alpha-MSH has protective role on ischemic renal injury and improves survival rates.

The Effect of alpha-MSH on Long-term Histologic Changes after Acute Ischemic Kidney Injury in Rats / 대한신장학회잡지

BACKGROUND: Ischemic acute renal failure (ARF) is increasingly recognized as involving chronic functional and structural sequelae. Ischemia reperfusion injury (I/R) plays a major role in delayed graft function and long-term changes after kidney transplantation, also. The present study was designed to evaluate the long-term changes after acute ischemic injury and whether renoprotection by alpha-MSH in the acute ischemic stage, could reduce the long-term sequelae. METHODS: In control group, ischemia/reperfusion injury was induced in male Sprague-Dawley rats by clamping Lt. renal pedicle for 15, 45, 60 minutes after removal of Rt. Kidney and followed by reperfusion. The animals in alpha-MSH group were injected alpha-MSH prior to reperfusion and then every day for 1 week. We measured BUN, creatinine at 24 hour after I/R injury, and in each group, 6 animals were sacrified at 1 week and 4 weeks after I/R injury to evaluate apoptosis, ED1, PCNA, and histopathologic changes. RESULTS: At 4 weeks after I/R injury, the remnant structural damage such as apoptosis, ED1 stained cells, MT (+) tubulointerstitial fibrosis were observed. alpha-MSH could reduce the initial functional injury, and apoptosis, ED1 stained monocyte, MT (+) tubulointerstitial fibrosis, also. CONCLUSION: Renal function was recoverd at 4 weeks after I/R injury, but structural sequelae such as apoptosis, fibrosis were remnant. alpha-MSH could attenuate initial functional damage and remnant fibrosis.

Inflammatory status in patients on chronic hemodialysis

All chronic renal failure patients show clinical and/or laboratory manifestations of inflammation. Prevalence of infection in chronic renal failure [CRF] patients is high due to immune dysfunction, also repeated cannulation and central venous catheters are probably responsible for the high incidence of infection. Detection of these pro-inflammatory cytokines in the plasma allow to detect patients with high risk of endotoxemia and also it was noticed that there is high degree of correlation between the inflammation, malnutrition and atherosclerotic process. The aim of the present study was to detect the level of plasma alpha-MSH as immunomodulator which counter acts the pro-inflammatory cytokines such as TNF-alpha, neopterin, nitric oxide [NO], C-reactive protein [CRP] and eridotoxins. This study was conducted on 40 subjects divided into three groups, 15 patients on maintenance hemodilaysis [GI], 15 patients with chronic renal failure not yet on dialysis [GII], and 10 healthy subjects as a control group [GIII]. Plasma alpha-MSH was measured using a double antibody radioimmunoassay, TNF-alpha and neopterin using specific enzyme-linked immunosorbent assays. Plasma nitrites were determined by a colorimetric method and endotoxin with the quantitative chromogenic method. The present study showed that most of the chronic renal failure patients show laboratory evidence of inflammation especially before starting dialysis. alpha-MSH was found to he elevated in both groups. Also, the levels of the proinflammatory cytokines, TNF-alpha, neopterin, nitric oxide, iNOS, C-reactive protein and plasma endotoxin were found to he elevated in both group I and group II. There was a positive correlation between alpha-MSH and endotoxemia. Also, there was a positive correlation between the level of endotoxins and the level of other pro-inflammatory cytokines suggesting the role of endotoxemia in stimulating the formation and release of the proinflammatory cytokines

Expression of alpha-melanocyte-stimulating hormone in human split-thickness skin graft / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the expression of alpha-melanocyte-stimulating hormone (alpha-MSH) in human split-thickness skin autograft and the role of alpha-MSH in hyperpigmented process of the grafted skin.</p><p><b>METHODS</b>Immunohistochemical technique was used to detect the expression and distribution of alpha-MSH in the split-thickness grafted skin and normal skin separately.</p><p><b>RESULTS</b>The expression of alpha-MSH in most of the split-thickness grafted skin was much stronger than the control skin. The positive ratio of alpha-MSH expression was 61.1% in the split-thickness grafted skin, 11.1% in the normal skin of the donor area and 16.7% in the normal skin around the recipient area. The expression of alpha-MSH in the split-thickness grafted skin was significant high, compared with the normal skins (P < 0.01). The expression of alpha-MSH in the normal skin of the donor area was no statistic remarkably differences compared to the normal skin around the recipient area.</p><p><b>CONCLUSION</b>The results indicate that the expression of alpha-MSH may markedly increase in the split-thickness grafted skin and correlate with its pigmentation after the skin graft. Overexpression of alpha-MSH may play an important role in hyperpigmented process of the skin graft.</p>

Drug Induced Cerebral Proerectile Effect in Anesthethetized Rats / 대한남성과학회지

PURPOSE: To examine the centrally elicited erectile effects of alpha-MSH, oxytocin, prostagladin E1, and anabolic steroids after intracerebroventricular administration. MATERIALS AND METHODS: We used anesthetized male Sprague-Dawley rats. After intracerebroventricular administration of normal saline (NS), alpha-melanocyte stimulating hormone (alpha-MSH), oxytocin acetate (OT), prostaglandin E1 (PGE1), methylprednisolone (MP), testosterone enanthate (TE) or 17beta-estradiol (E2) under stereotaxis, the intracavernosal pressure (ICP), systolic femoral artery pressure (FAP), heart rate (HR), time to first response, duration, and number of erectile responses and adverse reactions were evaluated for 60 minutes. To show whether erections were centrally elicited, the above criteria were re-evaluated after a bilateral pelvic neurotomy and bilateral orchiectomy. RESULTS: A cerebral proerectile effect was elicited only by alpha-MSH and OT with no significant changes of FAP or HR. With PGE1, significant changes in FAP and HR were observed. The ICP/FAP ratio was highest (0.49 0.03) with alpha-MSH. The mean time latency was shortest with OT (20.6 5.6 min). The mean duration was longest in alpha-MSH (11.6 8.7 min). The number of responses was highest with OT (3.6 0.7). Adverse reactions, such as stretching, yawning and ejaculation, were simultaneously observed during increases in ICP. In the cases of a bilateral pelvic neurotomy or bilateral orchiectomy, these elicited erectile responses disappeared. CONCLUSIONS: alpha-MSH had the most potent proerectile effect compared to OT and PGE1 as assessed by highest intracavernosal pressure as well as duration of erectile response. The results suggest that testosterone and the pelvic nerve were essential for the central proerectile response.

The Expression of the alpha-Melanocyte Stimulating Hormone (alpha-MSH) and Melanocortin-1 Receptor (MC1R) in the Epidermis of the Vitiligo / 대한피부과학회지

BACKGROUND: Vitiligo is a skin disease that is characterized by the loss of cutaneous pigmentation. alpha-Melanocyte stimulating hormone (alpha-MSH) is a neuroimmunomodulating peptide derived from proopiomelanocortin, and melanocortin-1 receptor (MC1R) is a surface receptor which is expressed by several other cutaneous cells including melanocyte and keratinocyte. Both of them have been known to be the main physiologic regulator for integumental pigmentation. OBJECTIVE: To evaluate the expression pattern of alpha-MSH and MC1R in the epidermis of vitiligo patients. METHODS: Specimens were obtained in lesional, perilesional and non-lesional skin in 10 patients with vitiligo and from 3 normal persons by the punch biopsy. And then, indirect immunofluorescence was done to show the pattern of expression of alpha-MSH and MC1R. RESULTS: Pattern of expression between alpha-MSH and MC1R was nearly the same. In vitiligo patients with stable disease state (7 of 10), the expression of alpha-MSH and MC1R in the non-lesional skin was more prominent than that in lesional area. In vitiligo patients with active disease state (3 of 10), the expression of alpha-MSH and MC1R in the lesional skin was more prominent than that in non-lesional area. CONCLUSION: Between the stable and active vitiligo patients, there was a different pattern of expression of alpha-MSH and MC1R in the lesional skin.

Altered Expression of Renal AQP1, AQP2 and NHE3 in Puromycin Aminonucleoside Induced Nephrotic Syndrome: Intervention by Alpha-MSH Treatment / 대한신장학회잡지

BACKGROUND: We examined whether puromycin aminonucleoside (PAN)-induced nephrotic syndrome (NS) is associated with altered renal handling of water and sodium along with changes of renal abundance of aquaporins (AQP1 and AQP2) and NHE3. Next we tested the effects of alpha-melanocyte stimulating hormone (alpha- MSH), a potent anti-inflammatory drug, on the PAN-induced renal functional derangement and the changes of renal AQPs and NHE3 abundance. METHODS: PAN was administered to Sprague-Dawley rats using two protocols: protocol 1 (180 mg/kg, single iv injection) and protocol 2 (100 mg/kg, single iv injection). RESULTS: In both protocols, PAN-induced NS was associated with decreased urine concentration, manifested by an increased urine output and decreased urine osmolality and TcH2O. Consistent with this, a marked downregulation of vasopressin-regulated collecting duct AQP2 expression was seen in PAN-induced NS. In protocol 2 where rats treated with moderate dose of PAN, alpha-MSH cotreatment prevented the reduction of urine osmolality and the increase of the FENa in the PAN-induced NS. This suggests that alpha-MSH may have protective effects against the renal functional deterioration induced by PAN. The renal abundance of the AQP1, AQP2 and NHE3 was reduced in PAN-induced NS in protocol 2, as seen in protocol 1. In contrast to the functional improvement, alpha-MSH cotreatment had marginal effects in the prevention of renal AQP1, AQP2 and NHE3 downregulation in PAN-induced NS. CONCLUSION: PAN-induced NS was associated with decreased urine concentration along with reduced renal AQP2, AQP1 and NHE3 abundance. Alpha-MSH may have protective effects against the renal functional deterioration (e.g., urine osmolality and FENa). However, alpha-MSH treatment alone is less likely to prevent the marked reduction of AQP2, AQP1 and NHE3 abundance in PAN-induced NS, in contrast to the previously known dramatic effects against the ischemia-reperfusion injury in kidney and small intestine.

The Effect of alpha-MSH on Tumor Necrosis Factor-alpha and NFkappaB Activation in Cultured Human Proximal Tubular Cells during Simulated Ischemia / 대한신장학회잡지

BACKGROUND: Tumor necrosis factor a (TNF), potent proinflammatory cytokine, may be related with ischemia/reperfusion injury induced tubular cell inflammation and apoptosis. We examined TNF and its major nuclear transcriptional factor, NFkappaB activation in cultured human tubular cells in hypoxic condition and the effect of alpha-MSH, potent antiinflammatory agent, which have been reported to reduce renal I/R injury in rats. METHODS: Hypoxic culture condition was produced by oxidative pathway inhibitor (Antimycin 2 mM) and glycolytic pathway inhibitor (deoxy-D- glucose 2 mM and 10 mM) for 1 hour and re-oxygenation was performed by placing the cells in normal medium. The expression of TNF mRNA was studied by RT-PCR and NFkappaB DNA binding activity was analysed by Electomobility shift assays (EMSA) and cellular apoptosis was assessed by the terminal deoxynucleotidyl transferase-mediated dUTP biotin nick-end labelling (TUNEL) method and DNA laddering. These data were compared between the alpha-MSH and the vehicle-treated groups. RESULTS: Measured ATP level was 49% of control by luciferase-based assay kit. I/R injury caused an increase in TNF mRNA and NFkappaB activation and was accompanied by morphological evidence of apoptosis. alpha-MSH significantly reduced the degree of apoptosis, as well as TNF mRNA and NFkappaB activity (TNF/L19 mRNA ratio, vehicle/alpha-MSH: 105.15 +/- 16.5/18.75 +/- 0.85, p<0.05) (NFkappaB activity, vehicle/alpha-MSH: 5624/4803 densitometric index (DI), p<0.05). CONCLUSION: These findings suggest that alpha-MSH can decrease cellular apoptosis in hypoxic tubular cells and this protective effect of alpha-MSH may be related, in partially, with supression of TNF and NFkappaB activity.