No abstract available.
alpha-MSH* ; Obesity

Adult ; Calcium-Transporting ATPases ; metabolism ; Erythrocyte Membrane ; metabolism ; Female ; Humans ; Hydrocarbons ; adverse effects ; Lipid Peroxidation ; Male ; Middle Aged ; Occupational Exposure ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Young Adult

The purpose of the present study was to investigate whether interleukin-2 (IL-2) changes the activity of sarcoplasmic reticulum (SR) Ca(2+) ATPase, sarcolemmal Ca(2+)ATPase and Na(+)/K(+) ATPase by measuring the Pi liberated from ATP hydrolysis with colorimetrical methods. It was shown that the activity of Ca(2+)ATPase in SR from IL-2-perfused (10, 40, 200, 800 U/ml) rat heart increased dose-dependently. After incubation of the SR with ATP (0.1 approximately 4 mmol/L), the activity of SR Ca(2+)ATPase increased dose-dependently in the control group. In the SR from 200 U/ml IL-2-perfused hearts, the activity of Ca(2+)ATPase was much higher than that in the control group. On the other hand, incubation of the SR with Ca(2+) (1 approximately 40 micromol/L) increased the activity of SR Ca(2+) ATPase in the control group. The activity of SR Ca(2+)ATPase of IL-2-perfused hearts was inhibited as the function to Ca(2+). Pretreatment with specific kappa-opioid receptor antagonist nor-BNI (10 nmol/L) for 5 min attenuated the effect of IL-2 (200 U/ml) on the activity of SR Ca(2+) ATPase. After pretreatment with pertussis toxin (PTX, 5 mg/L) or U73122 (5 micromol/L), IL-2 failed to increase SR Ca(2+)ATPase activity. The activity of SR Ca(2+)ATPase was not changed by incubation of SR isolated from normal hearts with IL-2. Perfusion of rat heart with IL-2 did not affect the activity of sarcolemmal Ca(2+)ATPase and Na(+)/K(+)ATPase. It is concluded that perfusion of rat heart with IL-2 increases the activity of SR Ca(2+)ATPase dose-dependently, which is mainly mediated by cardiac kappa-opioid receptor pathway including a PTX sensitive Gi-protein and phospholipase C. IL-2 increases the activity of SR Ca(2+)ATPase as the function to ATP, but inhibits the activity of SR Ca(2+)ATPase as the function to Ca(2+). IL-2 has no effect on the activity of sarcolemmal Ca(2+)ATPase and Na(+)/K(+)ATPase.
Animals ; Interleukin-2 ; pharmacology ; Male ; Myocardium ; enzymology ; Rats ; Rats, Sprague-Dawley ; Sarcolemma ; enzymology ; Sarcoplasmic Reticulum ; enzymology ; Sarcoplasmic Reticulum Calcium-Transporting ATPases ; metabolism ; Sodium-Potassium-Exchanging ATPase ; metabolism

OBJECTIVETo study the effect of cerebral mild hypothermia on cerebral mitochondrial ATPase activities in neonatal rats with hypoxic-ischemic brain damage (HIBD).

METHODSEighty-four seven-day-old Wistar rats were randomly assigned into four groups: sham-operated normothermic, sham-operated mild hypothermic, HIBD normothermic and HIBD mild hypothemic. HIBD was induced by left common carotid artery ligation, followed by 8% hypoxia exposure. At each time interval of 2, 6, and 12 hrs post-hypoxia-ischemia (HI), 7 rats were sacrificed and the brain tissues were sampled for detecting the activities of mitochondrial Na+K+ATPase and Ca2+ATPase.

RESULTSThe activities of mitochondrial Ca2+ATPase decreased significantly in the two HIBD groups compared with those of the two sham-operated groups at 2, 6, and 12 hrs post-HI. The HIBD mild hypothemic group had higher mitochondrial Ca2+ATPase activities compared with the HIBD normothermic group at 2, 6, and 12 hrs post-HI (5.25 +/- 0.61 micromol/mgPr.h vs 3.17 +/- 0.81 micromol/mgPr.h 4.59 +/- 0.81 micromol/mgPr.h vs 2.26 +/- 0.53 micromol/mgPr.h4.61 +/- 0.62 micromol/mgPr.h vs 1.31 +/- 0.78 micromol/mgPr.H, respectively) (P < 0.01). The activities of mitochondrial Na+K+ATPase decreased significantly in the two HIBD groups compared with those of the two sham-operated groups at 6 and 12 hrs post-HI. A significant difference was observed in the mitochondrial Na+K+ATPase activities between the HIBD mild hypothemic and HIBD normothermic groups at 6 and 12 hrs post-HI (5.25 +/- 0.66 micromol/mg Pr.h vs 3.76 +/- 0.78 micromol/mgPr.h, 4.74 +/- 0.80 micromol/mgPr.h vs 3.12 +/- 0.53 micromol/mgPr.h; P < 0.01).

CONCLUSIONSMild hypothermia following HIBD inhibits the decline in cerebral mitochondrial Ca2+ and Na+K+ ATPase activities in neonatal rats, thus providing protective effects against HIBD.

Animals ; Animals, Newborn ; Brain ; enzymology ; Calcium-Transporting ATPases ; metabolism ; Female ; Hypothermia, Induced ; Hypoxia-Ischemia, Brain ; enzymology ; therapy ; Male ; Mitochondria ; enzymology ; Rats ; Rats, Wistar ; Sodium-Potassium-Exchanging ATPase ; metabolism

Animals ; Brain ; drug effects ; metabolism ; Calcium-Transporting ATPases ; metabolism ; Female ; Mice ; Mice, Inbred Strains ; Polychlorinated Dibenzodioxins ; toxicity ; Sodium-Potassium-Exchanging ATPase ; metabolism

This study aimed to observe the general state and changes in pathophysiological indexes of multiple cerebral infarction rat model with Qi-deficienty and Blood-stasis syndrome. Rats were randomly divided into 4 groups(with 30 in each group): the normal group, the sham group, the model group and the Yiqi Huoxue recipe group. Rats in the model group and Yiqi Huoxue group were provided with interruptable sleep deprivation for 7 days before the multiple cerebral infarction operation, and followed by another 4 weeks of sleep deprivation; rats in the Yiqi Huoxue group were intragastrically administrated with drug at a dose of 26 g·kg⁻¹, once a day for 4 weeks. The general state was observed, and the pathophysiological indexes were measured at 48 h, 2 weeks and 4 weeks after administration. The results showed that rats in the normal group and the sham group represented a good general state and behaviors, with a normal morphological structure of brain tissues; rats in the model group featured yellow fur, depression, accidie, loose stools and movement disorder, with obvious brain histomorphological damage, which became aggravated with the increase of modeling time; rats in the Yiqi Huoxue group showed release in the general state and above indexes. Compared with the sham group at three time points, rats in the model group showed decrease in body weight, exhaustive swimming time and RGB value of tongue surface image, and increase in whole blood viscosity of the shear rate under 5, 60 and 150 S⁻¹, reduction in cerebral cortex Na⁺-K⁺-ATPase, Ca²⁺-ATPase activity and contents of 5-HT, rise in TXB2 levels and decline in 6-keto-PGF1a in serum(<0.05, <0.01). Compared with the model group, rats in the Yiqi Huoxue group showed alleviations in the above indexes at 2 w and 4 w(<0.05, <0.01). The results showed that the characterization and pathophysiological indexes in the multiple cerebral infarction rat model with Qi-deficiency and blood-stasis syndrome were deteriorated; Yiqi Huoxue recipe could significantly alliviate the abnormal conditions, which suggested of the model was stable and reliable and the pathophysiologic evolutionary mechanism might be related to energy metabolism dysfunction, vasoactive substance abnormality and changes in neurotransmitters.
Animals ; Calcium-Transporting ATPases ; metabolism ; Cerebral Infarction ; physiopathology ; Drugs, Chinese Herbal ; pharmacology ; Energy Metabolism ; Medicine, Chinese Traditional ; Qi ; Rats ; Sodium-Potassium-Exchanging ATPase ; metabolism

The aim of this research is to investigate the effects of paeoniflorin and menthol on the physiological function of Calu-3 cell membrane during the transport of puerarin. Calu-3 cell was used as the cell model to simulate nasal mucosa tissues, and the cell membrane fluidity, Na⁺-K⁺-ATPase activity and Ca²⁺-ATPase activity were detected by fluorescence recovery after photobleaching(FRAP) and ultramicro enzyme activity testing, in order to explore the mechanism of compatible drugs on promoting puerarin transport. The results showed that when puerarin associated with low, middle and high concentration of menthol or both paeoniflorin and menthol, the fluorescence recovery rate was increased significantly, while Na⁺-K⁺-ATPase activity had no significant change and Ca²⁺-ATPase activity was enhanced significantly as compared with puerarin alone. Therefore, it was concluded that menthol had the abilit of promoting the transport and the mechanism might be related to increasing membrane fluidity and activating Ca²⁺-ATPase.
Calcium-Transporting ATPases ; metabolism ; Cell Line, Tumor ; Cell Membrane ; Glucosides ; chemistry ; Humans ; Isoflavones ; metabolism ; Membrane Fluidity ; Menthol ; chemistry ; Monoterpenes ; chemistry ; Sodium-Potassium-Exchanging ATPase ; metabolism

BACKGROUND: The effects of melanocyte stimulating hormone(MSH) on the integument of many species, including mammals, are well known. The significance of MSH as a physiological regulator of cutaneous pigmentation in humans is still controversial. Although the administration of MSH results in skin darkening, previous reports suggest that cultured human melanocytes are relatively unresponsive to this peptide. This may be related to the conditions under which the melanocytes were cultured. OBJECTIVE: The purpose of this study was to investigate the effect of alpha-MSH on the morphological changes, survival, and melanization of cultured human melanocytes in a basal medium without any mitogen. METHOD: We examined the morphological changes, number and melanin contents of cultured human melanocytes in control(absence of alpha-MSH) and experimental groups(presence of 10(-8) M, 10(-7) M, and 10(-6) M alpha-MSH). RESULTS: 1. There were no significant morphological changes of cells between the control and experimental groups after 24, 48, and 72 hours' culture. The number and length of melanocyte dendrites showed no significant difference between the groups after 24, 48, and 72 hours' culture. 2. The number of melanocytes in the experimental groups(presence of 10(-7) M, and 10(-6) M alpha-MSH) were significantly higher than the number of melanocytes in control group after 72 hours culture(p<0.05). This effect of alpha-MSH was dose-related. 3. The melanin contents slightly increased in the experimental groups. The significant difference between the groups was showed in the presence of 10(-8) M alpha-MSH. CONCLUSIONS: alpha-MSH has no effect on the morphology, but increases the survival of cultured human melanocytes and has a melanogenic effect.
alpha-MSH* ; Dendrites ; Humans* ; Mammals ; Melanins ; Melanocyte-Stimulating Hormones ; Melanocytes* ; Pigmentation ; Skin

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.
alpha-MSH ; Biomarkers ; Cell Proliferation ; Humans* ; Melanins ; Melanocytes* ; Mitogens ; Monophenol Monooxygenase ; RNA, Messenger ; Up-Regulation

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.
Adenosine Triphosphate ; alpha-MSH* ; Animals ; Anoxia ; Apoptosis ; Biotin ; DNA ; Glucose ; Humans* ; Inflammation ; Ischemia* ; Rats ; RNA, Messenger ; Tumor Necrosis Factor-alpha*