RESUMEN
Objective:To explore the correlations of α-melanocyte stimulating hormone ( α-MSH) levels in serum and synovial fluid with progression of primary knee osteoarthritis (KOA). Methods:A retrospective analysis was conducted of the 96 patients who had been diagnosed as primary KOA at Department of Orthopedics, The First Hospital of Huizhou from October 2018 to October 2019. Radiographic severity of KOA was determined by Kellgren-Lawrence (K-L) grades; α-MSH levels were measured by enzyme-linked immunosorbent assay (ELISA). Levels of pro-inflammatory cytokine interleukin-1 β (IL-1 β) and matrix metalloproteinase-3 (MMP-3) were also detected. Another 64 patients with patellar dislocation, matched in age and gender, were enrolled as controls. The Numeric Pain Scale (NPS) and revised Oxford Knee Score (OKS) were employed to evaluate their symptomatic severity. Receiver operating characteristics (ROC) curve was used to compare α-MSH, IL-1 β and MMP-3 with regard to their diagnostic values in the K-L grading. Results:There were no statistically significant difference in age, gender and body mass index between the 2 groups, showing they were comparable ( P> 0.05). The α-MSH levels in synovial fluid were significantly lower in the KOA patients than in the controls [(16.9±3.8) pg/mL versus (18.8±2.7) pg/mL] ( P<0.001); there were no significant differences between the KOA patients and the controls in the serum α-MSH levels [(24.9±1.8) pg/mL versus (24.8±1.7) pg/mL] ( P>0.05). The α-MSH levels in synovial fluid were negatively correlated with K-L grades ( r=-0.382, P<0.001) and negatively correlated with NPS ( r=-0.382, P<0.001) but positively correlated with OKS ( r=0.339, P<0.001). Moreover, the α-MSH levels in synovial fluid were negatively correlated with the IL-1 β levels in synovial fluid ( r=-0.483, P<0.001) and with the MMP-3 levels in synovial fluid ( r=-0.336, P< 0.001). Conclusions:The level of serum α-MSH may not be correlated with the progression of KOA but the synovial fluid α-MSH is negatively correlated with the progression of KOA. Therefore, the expression level of α-MSH in joint synovial fluid can be used as a potential biomarker for assessment of severity of knee osteoarthritis.
RESUMEN
Objective To investigate the effect of α-melanocyte-stimulating hormone (α-MSH ) on the expression of mRNA and long noncoding RNA ( lncRNA ) in retinal vascular endothelial cells stimulated by hyperglycemia and hyperlipidemia. Methods The simian retinal vascular endothelial cells (RF/6A)were cultured and divided into normal control group,model control group,0. 1μmol/Lα-MSH group,0. 5μmol/Lα-MSH group and 1. 0μmol/L α-MSH group. The cells were stained with CM-H2 DCFDA to detect cell antioxidant capacity. The optimal concentration of α-MSH was screened. The cells from normal control group,model control group andα-MSH treatment group were collected at 24 hours after treatment,the total RNA was extracted,the cDNA library was constructed,and the high throughput RNA sequencing ( RNA-seq ) was carried out with bioinformatics analysis to analyze the expression profiling of mRNA and lncRNA. Results The fluorescence intensity of cells in 0. 5 μmol/L α-MSH group was significantly lower than that in model control group ( P<0. 05 ) .α-MSH of 0. 5μmol/L was chosen as the optimal concentration for subsequent experiments. Compared with the model control group, 243 mRNAs were significantly down-regulated,while 81 mRNAs were up-regulated in the α-MSH treatment group;53 lncRNAs were markedly up-regulated and 6 lncRNAs were down-regulated in the α-MSH treatment group. Bioinformatics analysis showed that the major enrichment pathways of the down-regulated genes were transforming growth factor-β( TGF-β) signaling pathway,focal adhesion signaling pathway and extracellular matrix ( ECM) receptor interactions pathways, and the main biological process involved was the regulation of small GTPase-mediated signal transduction. The co-expression gene enrichment pathways of differentially expressed lncRNA included ECM receptor interaction and hypoxia inducible factor-1(HIF-1) pathway,et al. These pathways were mainly involved in the biological processes, such as axon guidance and positive regulation of transcription from RNA polymerase Ⅱ promoter. Conclusions Under hyperglycemia and hyperlipidemia, the influence of α-MSH on the transcriptome of the retinal vascular endothelial cells manifests the downregulation of mRNA and upregulation of lncRNA. α-MSH may upregulate the lncRNA expression,which downregulates the downstream mRNA expression.
RESUMEN
Objective@#To investigate the effect of α-melanocyte-stimulating hormone(α-MSH) on the expression of mRNA and long noncoding RNA (lncRNA) in retinal vascular endothelial cells stimulated by hyperglycemia and hyperlipidemia.@*Methods@#The simian retinal vascular endothelial cells (RF/6A)were cultured and divided into normal control group, model control group, 0.1 μmol/L α-MSH group, 0.5 μmol/L α-MSH group and 1.0 μmol/L α-MSH group.The cells were stained with CM-H2DCFDA to detect cell antioxidant capacity.The optimal concentration of α-MSH was screened.The cells from normal control group, model control group and α-MSH treatment group were collected at 24 hours after treatment, the total RNA was extracted, the cDNA library was constructed, and the high throughput RNA sequencing (RNA-seq) was carried out with bioinformatics analysis to analyze the expression profiling of mRNA and lncRNA.@*Results@#The fluorescence intensity of cells in 0.5 μmol/L α-MSH group was significantly lower than that in model control group (P<0.05). α-MSH of 0.5 μmol/L was chosen as the optimal concentration for subsequent experiments.Compared with the model control group, 243 mRNAs were significantly down-regulated, while 81 mRNAs were up-regulated in the α-MSH treatment group; 53 lncRNAs were markedly up-regulated and 6 lncRNAs were down-regulated in the α-MSH treatment group.Bioinformatics analysis showed that the major enrichment pathways of the down-regulated genes were transforming growth factor-β(TGF-β) signaling pathway, focal adhesion signaling pathway and extracellular matrix (ECM) receptor interactions pathways, and the main biological process involved was the regulation of small GTPase-mediated signal transduction.The co-expression gene enrichment pathways of differentially expressed lncRNA included ECM receptor interaction and hypoxia inducible factor-1(HIF-1) pathway, et al.These pathways were mainly involved in the biological processes, such as axon guidance and positive regulation of transcription from RNA polymerase Ⅱ promoter.@*Conclusions@#Under hyperglycemia and hyperlipidemia, the influence of α-MSH on the transcriptome of the retinal vascular endothelial cells manifests the downregulation of mRNA and upregulation of lncRNA.α-MSH may upregulate the lncRNA expression, which downregulates the downstream mRNA expression.
RESUMEN
Aim To investigate the inhibitory effect of polyphenol from Cortex Mori( CMP) on melanogenesis in mouse melanoma B16 cells and its possible mecha- nism. Methods Melanoma B16 cells with high ex-pression melanin were induced by α-melanocyte-stimu-lating hormone ( α-MSH) to establish cell model. Cell viability was detected by MTT assay. The melanin syn-thesis and tyrosinase activity were measured by NaOH and L-Dopa assays, respectively. The tyrosinase (TYR), tyrosinase-related protein-1 (TRP-1), tyrosi-nase-related protein-2 ( TRP-2 ) and microphthalmia associated transcription factor ( MITF ) protein and mRNA levels were measured by Western blot and qRT-PCR, respectively. Results CMP could inhibit the melanin synthesis and tyrosinase activity in α-MSH stimulated B16 cells in a dose-dependent manner ( P<0.05) . The melanin content and tyrosinase activity significantly decreased by 52.95% , 32.85% at 20 mg ·L-1of CMP, respectively. Treatment of 100 mg· L-1of arbutin reduced the melanin content and tyrosi- nase activity by 17.29% , 16.75% , respectively. Based on the results of this study, CMP showed a stronger anti-melanogenesis activity than that of positive control arbutin. After treated by CMP, the protein and mRNA levels of TYR, TRP-1, TRP-2 and MITF were significantly inhibited compared to the α-MSH group ( P<0.05) . Conclusions CMP could suppress the melanogenesis in α-MSH stimulated B16 cells, and its mechanism may be related to its regulation of the pro-tein and mRNA expressions of TYR, TRP-1, TRP-2 and MITF, and the inhibition of tyrosinase activity.
RESUMEN
Background Reasearches showed that α-melanocyte-stimulating hormone (α-MSH) inhibits inflammation and ameliorates the ocular surface abnormalities in a scopolamine-induced dry eye rat model,and the managing effect of sodium carboxymethylcellulose (CMC) on dry eyes also has been determined.However,whether α-MSH can enhance the therapeutic effects of CMC remains to be investigated.Objective This study was to investigate the protective effects of α-MSH combined with CMC on ocular surface in a scopolamine-induced dry eye rat model.Methods Sixty clean female Wistar rats were randomly divided into normal control group,model control group,NaCl group,CMC group,α-MSH group and α-MSH+CMC group,and 10 rats for each group.The dry eye models were established by subcutaneous injection of scopolamine hydrobromide at 9:00,12:00,15:00 and 18:00 per day for 28 days.0.9％ NaCl solution,1×10 3 mg/ml α-MSH solution,0.5％ CMC eye drop,and 1 ×10-3 mg/ml α-MSH+0.5％ CMC solution were topically administered twice a day (8:00,17:00) since the initial day of modeling according to grouping.Shirmer Ⅰ test (S Ⅰ t),breakup time of tear film (BUT) and corneal fluorescence staining were performed before and 7,14,21,28 days after the application of drugs.At 28 days following the administration of drugs,the eyeballs of the rats were collected.Hemotoxylin and eosin staining was employed to examine the morphology of corneas,and periodic acid schiff (PAS) staining was used to count the conjunctival goblet cells.This study protocol was approved by Experimental Animal Ethic Committee of Tianjin Medical University (SYXK 2009-0001),and the use and care of the rats complied with ARVO Statement.Results The S Ⅰ t and BUT values were significantly reduced,and the corneal fluorescence staining scores were significantly increased over time following modeling in the model control group (all at P＜0.01).No significant differences were found in the S Ⅰ t,BUT and corneal fluorescence staining scores between model control group and NaCl group at various time points (all at P＞0.05).At 7,14 and 21 days after intervention,the S Ⅰ t values were (4.800±0.789),(4.100±0.516) and (4.300±0.856) mm in the α-MSH+CMC group,which were considerably higher than (2.875 ±0.719),(2.375 ±0.619) and (2.532±0.957)mm in the NaCl group (all at P＜0.01).At 7 days after intervention,the BUT values were (4.938± 1.843) seconds and (5.000±1.491) seconds in the α-MSH group and α-MSH+CMC group,which were significantly higher than (3.250±1.000) seconds in the NaCl group (both at P＜0.01).The corneal fluorescence staining scores in the CMC group,α-MSH group and α-MSH+CMC group were significantly lower than that in the NaCl group,with the lowest score in the α-MSH +CMC group (all at P＜0.05).The thickening of corneal epithelial layer,corneal edema and arrangement disorder of corneal stroma were found in the model control group and NaCl group;while slight corneal edema and epithelial cell proliferation were exhibited in the α-MSH+CMC group by hemotoxylin and eosin staining.PAS staining showed that the number of goblet cells was much more in the CMC group,α-MSH group and α-MSH+ CMC group than that in the model control group and NaCl group (all at P ＜ 0.01).Conclusions The sole application of α-MSH or CMC alleviates ocular surface damage and morphological abnormality to certain extent,and the combination of α-MSH and CMC generates more effective protection in comparison with sole administration of α-MSH or CMC.The early application of the drugs plays an improvement role in tear secretion and tear film stability in dry eyes.
RESUMEN
Objective To study the effects of α-melanocyte-stimulating hormone (α-MSH) on excessive inflammatory response of patients to traumatic brain injury (TBI) so as to prevent against the development of the secondary injury by observing the changes of α-MSH level in the serum of patients with TBI,and the relationships of the levels of serum α-MSH with the severity of TBI,and with the levels of tumor necrosis factor-α (TNF-α).Methods A total of 48 patients with acute TBI were divided into three groups according to GCS score:severe group with GCS 3-8 (n =18),moderate group with GCS 9-12 (n =16),and mild group with GCS 13-15 (n =14).Ten healthy volunteers were recruited as a control group.The blood samples were collected within 24 h and 3 d,5 d,7 days after injury.The concentrations of α-MSH and TNF-α in the separated serum were measured by double antibody sandwich enzyme-linked immunosorbent assay (ELISA).All variables were presented as ((x)±s).Repeated measures and analysis of variance and further multiple comparisons were carried out to compare variables.When necessary,the Student's t test was utilized.Pearson correlation analyses were performed to determine the correlations between variables.Results The serum α-MSH levels in the three TBI groups were lower than that in the control group (P ＜ 0.05).And the severer injury was,the lower α-MSH level was.The lowest α-MSH levels dropped to the trough on the 3rd day or the 5th day after TBI [severe group:(9.65 ±4.21) pg/mL,moderate group:(10.69 ±4.30) pg/mL,mild group:(18.89 ±7.19) pg/mLvs.control:(45.67 ± 10.95) pg/mL].While the serum TNF-α levels in three TBI groups were higher than that in the control group (P ＜ 0.05),and the TNF-α level was higher in the severer group.The peak values of TNF-α in the three TBI groups reached on the 3rd day after TBI [severe group:(37.24 ± 18.28) pg/mL,moderate group:(26.19 ±6.78) pg/mL,mild group:(18.60 ±7.83) pg/mL vs.control:(10.74 ± 1.71) pg/ mL].There were negative correlations between the levels of serum α-MSH and TNF-α at four intervals.Conclusions In patients with TBI,the serum levels of α-MSH decreased,and the lowest levels of α-MSH dropped to the trough on the 3rd day or the 5th day after TBI.While the levels of TNF-α increased,and the peak values reached on the 3rd day after TBI.And as the injury was more severe,these changes were more significant.There were negative correlations between the serum α-MSH levels and TNF-α levels in general.
RESUMEN
Background Retinal neovascularization (RNV) occurs in multiple eye diseases,which can lead to bleeding and retinal detachment.Therefore,inhibition of pathological RNV is becoming crucial to the treatment of ocular diseases.Research has shown that α-melanocyte-stimulating hormone (α-MSH) inhibits retinal angiogenesis during physiological development;however,the effects of α-MSH on pathological RNV remain unknown.Objective This study was to investigate the effects of intravitreal injection of α-MSH at different concentrations on pathological RNV in a mouse model of oxygen-induced retinopathy (OIR).Methods Forty healthy clean C57BL/6J mice were randomly divided into OIR+0.33 μg/μl α-MSH,OIR+ 1.67 μg/μl α-MSH,OIR+3.30 μg/μl α-MSH,OIR,and normal control groups at postnatal day 7 (P7),with 8 pups in each group.The α-MSH intervention groups and OIR group were exposed to high oxygen (75 ±2)％ for 5 days,then maintained under normal air condition for another 5 days;whereas the normal control group was raised under normoxia for 10 days.Retro-orbital injection of high molecular weight fluorescein isothiocyanate-dextran (FITC-dextran) was performed on P17 mice.The retina whole mounts were prepared to reveal retinal vasculature and quantify relative area of vessel obliteration.The mouse eyeballs were subjected to paraffin sections and hematoxylin-eosin staining,and the average number of pre-retinal nuclei per section was quantified.Results FITC-dextran labeled retinal whole mounts showed that the relative vessel obliteration area in normal control,OIR,OIR+0.33 μg/μl α-MSH,OIR+ 1.67 μg/μl α-MSH,and OIR+3.30 μg/μl α-MSH groups were (0.00±0.00) ％,(23.01 ±3.39) ％,(18.14±7.20) ％,(15.64±7.07) ％,and (7.62±6.52) ％,respectively.There was a statistical significance in the relative avascular area among the groups (F=19.635,P＜0.05).The relative avascular area in the OIR group was significantly higher than that in the OIR+3.30 μg/μl α-MSH group (t=4.293,P＜0.01).The results of histopathological examinations showed that the average number of pre-retinal nuclei per section in normal control,OIR,OIR+0.33 μg/μl α-MSH,OIR+1.67 μg/μl α-MSH,and OIR+3.30 μg/μl α-MSH groups were 0.00±0.00,11.45 ±4.26,6.35 ±2.34,4.96 ± 1.79 and 1.03 ± 1.25,respectively.There was a statistical significance in the average number of pre-retinal nuclei per section among the groups (F =147.87,P＜0.05).The average number of pre-retinal nuclei per section in the OIR group was significantly higher than that in the OIR+0.33 μg/μl α-MSH,OIR+ 1.67 μg/μl α-MSH,and OIR+3.30 μg/μl α-MSH groups,the differences between the groups had statistical significances (all at P＜0.001).Conclusions α-MSH reduces the relative area of vessel obliteration and the average number of pre-retinal nuclei in the retinas of OIR mouse model.The inhibitory effects of α-MSH on the pathological RNV are dose-dependent.
RESUMEN
Arquate nucleus, a convergence site of peripheral and central signals, plays a fundamental role in the control of food intake. Orexigenic neurons that secrete neuropeptide Y (NPY) and Agouti-related peptide (AgRP) and anorexigenic neurons secreting Pro-opiomelanocortin (POMC) are involved in this action. Both groups of neurons respond to peripheral signals such as insulin and leptin and are reciprocally inhibited. alpha Type melanocyte stimulating hormone (alphaMSH), liberated by POMC neurons, reduces food intake activating melanocortin receptor 4 (MC4R), located in second order neurons of the paraventricular nucleus. NPY/AgRP antagonize the effects of this peptide on MC4R receptors,maintaining an inhibitory tone on áMHS liberation, mediated by the activation of gabaergic receptors of POMC neurons. The study of these mechanisms will allow the development of new medications, especially MC4R agonists, to reduce nutrient intake...
Asunto(s)
Humanos , Ingestión de Alimentos/fisiología , Ingestión de Energía/fisiología , Obesidad/metabolismo , /fisiología , Hormonas Estimuladoras de los Melanocitos/fisiología , Proopiomelanocortina/fisiologíaRESUMEN
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.
Asunto(s)
Animales , Ratas , Lesión Renal Aguda , alfa-MSH , Quimiocina CCL5 , Creatinina , Interleucinas , Isquemia , Riñón , Hígado , Pulmón , Modelos Animales , Monocitos , Nefrectomía , Insuficiencia Renal Crónica , ReperfusiónRESUMEN
Objective To study the effect of alpha-melanocyte stimulating hormone (α-MSH) and its novel analogue ( STY39 ) on the production of tissue factor ( TF ) and tissue factor pathway inhibitor (TFPI) stimulated by lipopolysaccharide (LPS) in primary mouse brain microvascular endothelial cells (MBMECs). Methods Female BALB/c mice were selected,purified and primarily cultured for 5 to 7 days. Immunofluorescence assay was use to detect the Ⅷ factor related antigen and identify the MBMEC model. The MBMECs were divided into eight groups:PBS control group, LPS stimulation group, after LPS stimulation 1,2,and 3 h adding 10 -7 mol/Lα-MSH groups or STY39 group (LPS+α-MSH,LPS+STY39) ( n=4 wholes in each group) . The cell culture supernatant and cells were collected at 6 and 8 h after LPS stimulation. An enzyme-linked immunosorbent assay was used to detect the concentrations of TF and TFPI in cell supernatant. RT-PCR was used to detect the expression levels of TF mRNA. Results (1) LPS could induce MBMEC to produce TF and TFPI proteins. The level of TF in the cell culture supernatant reached the peak at 6 h,and the level of TFPI reached the peak at 8 h. (2) At 1,2,and 3 h after LPS stimulating MBMEC,10 -7mol/L α-MSH or STY39 were given. They could significantly decrease the TF protein content in the cell supernatant (P<0. 01),especially the effects of giving α-MSH or STY39 were most significant at 1 h after LPS stimulation (P<0. 05). The effect of STY39 for decreasing TF content was more significant than that of α-MSH (P<0. 05);however,α-MSH and STY39 did not have significant up-regulating effects for LPS inducing MBMEC to produce TFPI. (3) After LPS stimulation,10 -7 mol/Lα-MSH or STY39 were given at different time points. They significantly down-regulated the expression level of MBMEC TF mRNA (P<0. 01). The effect was most significant at 1 h time point (P<0. 05),but there was no significant difference in the effects betweenα-MSH and STY39. Conclusion Bothα-MSH and STY39 can suppress LPS-induced primary MBMEC to produce TF protein and express TF mRNA,and the effect of administration is better after 1 h LPS stimulation. The suppressive effect of STY39 on the production of TF protein is superior toα-MSH.
RESUMEN
Objective To evaluate the effect of α-melanocyte stimulating hormone (α-MSH) and its novel analogue STY39 on the production of tissue factor pathway inhibitor (TFPI) in mice with endotoxemia.Methods Female BALB/c mice were randomly divided into eight groups with 9 mice in each group.Endotoxemia was reproduced by intraperitoneal injection of lipopolysaccharide (LPS,25 μg/kg) and D-galactosamine (D-Gal,100 mg/kg).The animals of the control group were given phosphate buffered solution (PBS) instead.In the experimental groups,the mice were injected intraperitoneally with 2.5 mg/kg α-MSH or STY39 at 1,2 or 3 hours following LPS injection.The orbital blood was collected at different time points,and tissues of lung,liver,and kidney were collected 8 hours after the administration of LPS.The plasma TFPI levels were determined by enzyme linked immunosorbent assay (ELISA),and the expression of TFPI mRNA in different tissues was determined with reverse transcription-polymerase chain reaction (RT-PCR).Results The plasma TFPI levels (μg/L) began to increase (11.84 ± 1.55) in the endotoxemia mice 4 hours after LPS challenge and reached the peak (23.49 ± 1.12) at 8 hours.α-MSH or STY39 treatment at 1,2 or 3 hours after LPS challenge could significantly increase the TFPI content,with the best drug effect at 1 hour after LPS challenge (the blood was collected 8 hours after LPS challenge,α-MSH group:58.79 ± 2.67 vs.28.49 ± 1.69,STY39 group:71.08 ± 2.13 vs.28.49 ± 1.69,both P＜0.01),and the effect of STY39 was better than that of α-MSH (P＜0.01).A small amount of TFPI mRNA expression was observed in each tissue of the healthy mice.After LPS challenge,TFPI mRNA expression was increased in all the tissues,especially in the lung,liver and kidney.α-MSH or STY39treatment at 1 hour after LPS challenge could significantly up-regulate the expression of TFPI mRNA in the lung and liver (A value,α-MSH in lung:51.10 ±2.89 vs.32.43 ±2.51,STY39 in lung:72.11 ±3.48 vs.32.43 ±2.51;α-MSH in liver:43.21 ± 2.12 vs.29.29 ± 2.06,STY39 in liver:66.82 ± 1.76 vs.29.29 ± 2.06,both P＜0.01).The treatment with STY39 at 1 hour after LPS challenge could significantly up-regulate the expression of TFPI mRNA in the kidney (A value:45.21 ± 1.80 vs.30.44 ± 2.23,P＜0.01),but the treatment with α-MSH had no obvious effect (A value:24.61 ± 1.98 vs.30.44 ± 2.23,P＞0.05).The enhancing effect of early administration of STY39 on TTPI mRNA expression in the lung,liver and kidney tissues of endotoxemia mice was more powerful than that of α-MSH (all P＜0.01).Conclusion The early administration of α-MSH or STY39 may up-regulate TFPI production in the mice with endotoxemia,and the effect of STY39 is superior to α-MSH.
RESUMEN
Raw264.7 cells were incubated with receptor activator of NF-kappa B ligand (RANKL) and α-melanocyte stimulating hormone(α-MSH) for6 d.The amount of osteoclast cells were counted by tartrate resistant acid phosphatase staining and the acid phosphatase activity was assayed.The expressions of 5 melanocortin receptors (MCR) in Raw264.7 cells were determined by RT-PCR.The results showed that the number of osteoclasts in RANKL +α-MSH group was significantly increased compared with RANKL group (P ＜ 0.05),but there was no osteoclast formation in α-MSH group.Compared with control group and α-MSH group,the acid phosphatase activities were significantly increased in RANKL group and α-MSH+RANKL group (P＜0.05).All five MCRs were expressed in the Raw264.7 cells shown by RT-PCR.These results suggest that α-MSH may promote osteoclasts formation through RANK signaling pathway.
RESUMEN
Background The excitotoxicity to retinal neurons caused by abnormal elevation of glutamate in retina is a common pathology concomitant with major blind-causing eye diseases.However,an effective approach to protect retinal neurons from glutamate-induced excitotoxicity is still lack.Intraperitoneal administration of α-melanocyte stimulating hormone(α-MSH)has been shown to protect hippocampal neurons from glutamate-induced excitotoxicity.Objective This study was to investigate the protective effect of α-MSH on glutamate-induced excitotoxicity in a chicken embryonic retinal explant culture system.Methods The retinas were isolated from chick embryos at embryonic day 9(E9) and cultured as explants.The explants at 3,5 and 7 days in vitro and the retinas at corresponding embryonic day 12,14 and 16(E12,E14,E16)were collected.The morphology of explant cultures was examined by hematoxylin and eosin staining,and the expression of melanocortin receptors (MCRs)was analyzed by real-time PCR.In the experiment of glutamate-induced excitotoxicity,the retinal explants at 4 days in vitro were treated with glutamate for 48 hours,α-MSH was incubated with the explants 30 minutes before and during the glutamate treatment period.Then the apoptotic cells were detected by TUNEL staining and quantified.The glutamate alone treated-explants and those treated with culture media were included as controls.The expression of glial fibrillary acidic protein(GFAP) at 48 hours after treatment in all retinal explants was analyzed by real-time PCR.Results Hematoxylin and eosin staining showed that the retinal explants exhibited similar morphology to those observed in the retinas from chick embryos at the corresponding developmental stages.The real-time PCR analyses of chick retinas showed that MC1R mRNA level at E9,E12,E14 and E16 was significantly lower than that in post-hatch day 1 (all P=0.000) ;whereas the transcript level of MC5R was significantly increased from E9 to E12 and E14 (both P =0.000),and then gradually decreased from E14 to P1.The expression of these genes showed similar temporal patterns in the retinal explant cultures.TUNEL staining revealed that treatment of the retinal explant cultures with α-MSH substantially and significantly reduced number of apoptotic cells induced by glutamate (P =0.000),which was accompanied by significant suppression of glutamate-induced GFAP up-regulation (P =0.000).Conclusions Application of α-MSH dramatically ameliorated glutamate-induced cell death in retinal explant cultures.This protective effect may be due to α-MSH-mediated suppression of astrogliosis caused by abnormal elevation of glutamate.
RESUMEN
Objective To detect the variations of the serum α-MSH and TNF-α in multiple-trauma patients and discuss their role in severity of casualties.Methods Fifty casualties were divided into two groups for study.There were 30 casualties with moderate severe trauma(ISS 16 ～ 25 point)and 20 patients with extreme severe trauma(ISS ＞ 25 point),and another 15 healthy subjects were enrolled as controls.The blood samples were obtained within 24 hours,and 3 days,5 days,7 days after admission.The serum levels of α-MSH and TNF-α in casualties with multiple injuries were determined by using enzyme-linked immunosorbent double antibody sandwich method(ELISA).The data were expressed in((x)± s),and analyzed with chi-square test and repetitive measures of ANOVA by using SPSS 13.0 package.P value less than 0.05 indicated statistical significance Results The serum α-MSH levels of casualties within 24 hours,and 3 days,5 days,7 days after injury in the two groups were much lower than those in the control group (P ＜ 0.01),while the serum TNF-α levels of casualties were much higher than those in the control group (P ＜0.01).The serum α-MSH levels of casualties with extreme severe traumawere lower,and the TNF-αlevels of casualties with extreme severe trauma were higher than those in patients with moderate severe trauma(P ＜0.01,respectively).There were negative correlations between two biomarkers 24 hours,5d and 7d after injury.Conclusions In casualties,the serum levels of α-MSH decreased and the serum levelsof TNF-α increased,and the degrees of changes were closely depended on the severity of trauma,the more severe the more significant changes.There was a negative correlation between two biomarkers.
RESUMEN
Objective To evaluate the effect of this neuroendocrine hormone on protein expression by treating the human dermal fibroblasts with a-melanocyte stimulating hormone (α-MSH ).Methods Thehuman dermal fibroblasts was cultured, and the total protein of the fibroblasts were separated with immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE). After Coomassie bright blue staining, gel images were acquired by Image-scanner and then analyzed with the PDQuest software. 2-DE maps of fibroblasts were established. Partial differently expressed protein spots were incised from gels and digested by trypsin in-gel. Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and MSDB database searching by Mascot? software were used for protein identification. Results Well-resolved, reproducible 2-DE patterns of dermal fibroblasts treated with and without crMSH were obtained. 8 differently expressed protein spots were detected, among which 8 obtained peptide mass fingerprints (PMF) by MALDI-TOF-MS analysis. Among these proteins, of particular interest were five proteins annexin I, HSP27 and lamin A, etc. Conclusions Proteins expressed by human dermal fibroblasts treated with or without crMSH are different, and some of the differently expressed proteins involve apoptosis, intracellular signal transduction and framework construction and so on, which may be associated with anti-fibrosis effects of (a)-MSH on human dermal fibroblasts.
RESUMEN
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.
Asunto(s)
Animales , Ratas , alfa-MSH , Apoptosis , Muerte Celular , Técnicas de Cocultivo , Citocinas , Supervivencia de Injerto , Insulina , Ionomicina , Trasplante de Islotes Pancreáticos , Islotes Pancreáticos , Óxido Nítrico , Factor de Necrosis Tumoral alfaRESUMEN
Cutaneous dyschromia is a common problem of plastic and cosmetic surgery. There are not good methods to resolve this problem in present. The mechanisms of skin pigmentation process are not well understood. It is now recognized the correlation between melanogenesis and alpha-melanocyte-stimulating hormone. Alpha-msh enhances activity and quantity of tyrosinase as well as stimulates melanogenesis, specifically to stimulate eumelanin synthesis by activating the melanocortin-1 receptor (MC-1R) on melanocytes. Alpha-msh stimulates melanocyte prolixferation, formation of dendricity, and transfer of melanosomes to keratinocytes, It also protects melanocytes from the damaging effects by immunomodulation. Also, alpha-msh interact with other factors to affect pigmentation indirectly. Agouti signaling protein(ASIP) is a endogeneous antagonist of alpha-msh can induces pheomelanin synthesis by competing with alpha-msh for binding to the MC-1R.Follwing better understood the mechanisms of skin pigmented process, it will provide effective methods to resolve acquired human cutaneous hyperpigmentation and hypopigmentation.
RESUMEN
Objective:To study the influence of agouti signal protein(ASIP) on melanocyte function in skin autograft,so as to understand the cause of hyperpigmentation in skin autograft.Methods: Guinea pigs were used to establish a skin autograft hyperpigmentation model.The skin autografts in model animals were injected with ASIP or normal saline(control).RT-PCR technique was used to detect the tyrosinase mRNA expression in melanocytes of skin autografts and Masson-Fontana staining technique was used to detect the melanin contents in skin autografts in ASIP treatment group;and the results were compared with those of control group(n=13) and normal guinea pigs(n=5).Results: The expression of tyrosinase mRNA and the melanin content in skin autografts in ASIP treatment group were both lower than those of control group and normal guinea pigs((P
RESUMEN
Estrogen and progesterone are thought to be responsible for the pigmentary changes in pregnancy and also melasma. To investigate the action mechanism of estrogen and progesterone on the facultative skin pigmentation, Human melanocytes and keratinocytes were cultivated in the forms of pure melanocyte culture, co-culture (melanocytes and keratinocytes were cultivated together in a vessel but they were separated with membrane) or mixed culture (melanocytes and keratinocytes were cultivated together in a vessel in mixed form). After 2 days of cultivation in the presence of hormones (estrogen, progesterone and melanocyte stimulating hormone), the author studied the cell proliferation, the cellular features (the number of dendrites, perimeter and area), and the tyrosinase activity of melanocytes. Progesterone or melanocyte stimulating hormone increased in both the cell growth and the tyrosinase activity in pure melanocyte culture but estrogen did not. However, mixed culture treated with estrogen lead to increases in the tyrosinase activity. Pure melanocyte culture treated with estrogen or progesterone increased in the cell perimeter and the area but not in the number of dendrites. Co-cultured melanocytes without hormones revealed more increases in the perimeter (p.0.01) and the area (p.0.01) even in the number of dendrites (p.0.01) compared to the pure cultured melanocytes treated with the hormones. It was postulated with these results that estrogen, progesterone and keratinocyte possibly induced hyperpigmentation of the skin via the keratinocytes stimulated by estrogen, via the proliferation of melanocytes induced by progesterone, and via the cellular features altered by keratinocytes.
Asunto(s)
Humanos , Embarazo , Proliferación Celular , Técnicas de Cocultivo , Dendritas , Estrógenos , Hiperpigmentación , Queratinocitos , Melanocitos , Melanosis , Monofenol Monooxigenasa , Progesterona , Pigmentación de la Piel , PielRESUMEN
Apoptosis frequently occurs in acute renal injury but molecular mechanisms responsible for this distinct form of cell death are largely unknown. Fas belongs to the TNF/nerve growth factor superfamily and engagement by Fas ligand induces apoptosis in various epithelial cells. To investigate the role of apoptosis and associated molecular mechanisms, we examined the occurrence of apoptosis and Fas expression as well as the therapeutic effect of alpha-MSH, a potent anti-inflammatory cytokine in ischemic ARF rat model as well as its effect on Fas expression. The expression of Fas was studied by western blot analysis and semiquantitative RT-PCR. Apoptosis was assessed by the TUNEL method and the degree of apoptosis and Fas expression, as well as biochemical, histological data were compared between the alpha-MSH and the vehicle treated groups in 40 minute renal artery clamping ischemic ARF rat models. Intraperitoneally administered alpha-MSH significantly reduced renal injury, measured by plasma blood urea nitrogen, creatinine level and the degree of tubular necrosis(106.5+/-13.3/54.7+/-5.45mg/dL for BUN, 1.77+/-0.29/1.03+/-0.06mg/dL for creatinine 24 hours after ischemia)(p=0.003, p=0.01), (5.4+/-1.94/2.6+/-0.7 for injury score 24 hours after ischemia)(p=0.01). Ischemia caused significant upregulation of Fas mRNA and protein and was accompanied by morphological evidence of apoptosis. alpha-MSH significantly reduced the degree of apoptosis, as well as Fas[(mean apoptotic cell : 23.7+/-12.5/11.0+/-5.7 per 200 field at 4 hours after ischemia(p=0.04), 31.6+/-24.7/18.1+/-11.5 per 200 field at 24 hours after ischemia(p=0.25)]. (Fas protein expression : sham : 1409+/-355DI(densitometric index)) 2818.3+/-1100/1306+/-643.4DI at 24 hours and 5541.5+/-1597.5/ 2866.7+/-788.9DI at 72 hours after ischemia)(p=0.07, 0.047). These results suggest that Fas upregulation induced tubular cell apoptosis may contribute to the pathogenesis of ischemic ARF and the beneficial effect of alpha-MSH is partially mediated by these inhibitory effects on Fas system.