The changes of α-melanocyte-stimulating hormone in the serum of patients with TBI / 中华急诊医学杂志

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.

Inhibitory effect of alpha-melanocyte stimulating hormone and its novel analogue on the production of tissue factor in mouse brain microvascular endothelial cells / 中国脑血管病杂志

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.

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.

The change of α-melanocyte-stimulating hormone(α-MSH)in the serum of patients multipleinjuries / 中华急诊医学杂志

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.

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.

Effects of alpha-Melanocyte Stimulating Hormone on Apoptosis and Fas Expression in Ischemic Renal Injury in Rats / 대한신장학회잡지

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.