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AIM: To investigate the differentiation from human mesenchymal stem cells(hMSC) into neuron-like cells by the increase in intracellular cyclic AMP. METHODS: hMSC were separated from human marrow with Ficoll-Paque reagent and expanded in culture medium. hMSC were induced to differentiate into neurons with Forskolin and 3-isobutyl-1-methyl-xanthine (IBMX). Neuron-specific enolase(NSE), neurofilament(NF), glial fibrillary acaidic protein(GFAP) were detected by immunohistochemistry. RESULTS: hMSC were expanded as undifferentiated cells in culture for more than 10 passages. Forskolin/IBMX can induce hMSC to exhibit a neuronal phenotype, expressing NSE and NF-M in 5 hours. But the neuron-like cells didn't express the glial astrocyte marker GFAP. CONCLUSION: hMSC can be induced to differentiate into neurons by increase in the intracellular cAMP.
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AIM: The effects of Jiere Xingshen(JRXS) Injection on cAMP, IL-1 ? content in hypothalamus (HP) of endotoxin(ET)-induced feverish rabbits were studied. METHODS: The ET-induced fever model was established in rabbits and the cAMP content in hypothalamus (HP) and csf, IL-1 ? content in HP were determined by radioimmunoassay following intravenous infusion of JRXS. RESULTS: In ET group, the ?T [(0 40?0 11)℃], TRI 1(1 78?0 79), cAMP content in HP [(2 90?0 40) nmol/g], cAMP content in csf [(32 10?4 51) nmol/L)], IL-1 ? content in HP[(6 08?0 79) ng/g] were higher than that of NS and JRXS+ET group ( P
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0.05);but statistical significance was found between the ?-hGH group with rhGH+GPT group and CPT group (P 0.05).Conclusions The results demonstrate that ?-hGH had no influence on the level of CEA,cAMP and cGMP of the transplanted colonic COLO-320 cell line carcinoma in nude mice.
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Objective To explore the mechanism of Shema Zhichuan Liquid (SZL) for asthma. Methods Forty rats were allocated to normal control group (Group A), model group (Group B),western medicine group (Group C) and SZL group (Group D). Radioimmunoassay was used to observe the levels of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) and the ratio of cAMP/cGMP.Results After treatment, cAMP level and cAMP/cGMP ratio were increased and cGMP level decereased in Group C and Group D as compared with Group B (P
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To investigate the interaction of stimulatory GTP binding protein (G(s)) pathways with others, we overexpressed wild type alpha subunit of G(s) (G(s) alpha), constitutively activated R201E G(s) alpha, and dominant negative G226A G(s) alpha in COS-1 cells by transfection with DEAE-dextran, respectively, The expression of various G proteins in the transfected cells was analyzed after 72 h by quantitative Western blots, and cAMP production by stimulation with isoproterenol and forskolin was quantitated using cAMP binding proteins, The expression of Gs alpha increased about 5-fold in the transfected cells, with concomitant increase in the small forms. However, there was no significant alteration the in the level of the alpha subunit of inhibitory G protein (G(i)) and G(q), and the beta subunits of G proteins. The cAMP level without stimulation increased in the cells transfected with G(s) alpha regardless to the type of mutation, Treatment with either isoproterenol or forskolin resulted in comparable increase of the cAMP level in all the transfected cells, though the ratio to its respective basal level was smaller in the G(s) alpha-transfected cells, From this experiment, we found that the expression of the other G proteins and the signaling pathway producing cAMP did not change significantly by transiently expressing wild type, constitutively activated type, and dominant negative type of G(s) alpha. Analysis of the effects of long-term expression of Gs alpha would contribute to better understanding on how the G(s) alpha signaling system interacts with other signaling pathways and how it adapts to the changed environments.
Subject(s)
Animals , Blotting, Western , Carrier Proteins , Colforsin , COS Cells , Cyclic AMP , DEAE-Dextran , GTP-Binding Proteins , Isoproterenol , Protein Engineering , Recombinant Proteins , TransfectionABSTRACT
Nasopharyngeal carcinoma (NPC) in BALB/c nude mice (NM) was re-produced by hypodermic implanting human CNE-2 cell to the right-side of neck-backposi-tion. The changes of plasma malondialdehyde (MDA) and cAMP/cGMP ratio in NM bea-ring human NPC were examined. Meanwhile, we observed that tumor's size, plasma MDAand cAMP/cGMP ratio in NM bearing human NPC were affected by polysacchoridopetide(PSP). Results indicated that in comparison with the control group, NM bearing humanNPC showed a marked increase in plasma MDA(18.25?1.37 nmol/ml),cAMP/cGMP ratio(4.55?0.35), a marked decrease in plasma, cAMP (71.83?8.51 pM/ml) and cGMP(15.77?0.65 pM/ml), both high or low-concentration PSP inhibited NPC growth.Rate of inhibition was between 59.58 to 95.53 percent. PSP decreased plasma MDA,elevated cAMP, cGMP's levels and recovered cAMP/cGMP ratio. Our results suggestedthat the elevated level of MDA in NM bearing human NPC was closely associated withthe presence of NPC. PSP had antitumor effect, probably by means of decreasing freeradicals and recovering cAMP/cGMP ratio.
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In order to determine further whether cAMP and PGE takes part in the mechanism of EGTA-induced fever in the present experiment with 40 New Zealand rab bits, the influence of intravenous infusion of sodium salicylate (SS) on the febrile response and the rise of cAMP and PGE_2 concentration in c.s.f. produced by intraventricular perfusion of EGTA were observed. The results showed that intravenous infusion of SS (9.6?M/min. following an initial dose of 1.25mM) could inhibit the rise of PGE_2 concentration in c.s.f. (P0.05) after 0.6 mM EGTA was perfused into lateral cerebroventricle. The concentration of cAMP in c.s.f. was correlated apparently positively with the febrile response in EGTA-induced fever (r=0.9839, P
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Fever model was made by intravenously injected leucocytic pyrogen(LP,EP). Chai Ge Jie Ji Tang CGJJT fluid was perfused into rabbits stomach by mouth. 65 New Zealand white rabbits were divided into four groups: (1) control group; (2) LP group; (3) CGJJT fluid group; (4) LP+ CGJJT fluid group. Effects of CGJJT fluid on LP fever and cAMP concentration in cerebrospinal fluid (CSF)were observed. The results obtained from our experiments showed that: (1) The rectal temperature of normal rabbits markedly affected by CGJJT fluid. (2) Febrile response of LP was significantly inhibited by CGJJT fluid (P
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The levels of cyclic AMP and prostaglandin E (PGE) in the brain are raised in many kinds of fever induced by various pyrogens, we also know when the ratio of Na~+/Ca~(++) is changed in the brain, the body temperature changes. So it is suggested that cAMP, PGE and the ratio of Na~+/Ca~(++) are major central mediums in febril response. It is not clear what relationship is among these three factors and what are their roles respectively in the fever mechanism? It must be very helpful for the development of fever study if we can have them correctly answered. In this paper, we change the Na~+/Ca~(++) ratio by intraventricular perfusion with EGTA or CaCl_2, and observed the temperature response, levels of cAMP and PGE_2 in c.s.f. It is found that there was a positive correlation between cAMP and the body temperature(r=0.9913, P0.05). EGTA can raise body temperature. But CaCl_2 pretreatmentd could lower the EGTA fever.
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Intravenous injection of milk in dose of 0.6ml/kg into rabbits produced fever, putting an acupuncture needle into "Baihui (Hundred Flower) point" and gave intermittent electric stimulations. The thermal response peak (△T) of fever fell, the thermal response index (TRI) reduced and body temperature recovered fast from fever. The evidence showed that electric acupuncture can inhibit this type of fever. The contents of cAMP and L-enkephalin (LEK) in cerebrospinal fluid before fever, at the intervals of 2 and 3.5 hours in fever were determined by radioimmunoassay (RIA), and the resuIts indicated the central concentrations of cAMP in fever increased significantly, as well as the central concentrations of LEK increased at interval of 2 hours, but recovered at interval of 3.5 hours. While electric acupuncture inhibited fever of rabbits, it reduced central concentrations of cAMP and increased the central concentrations of LEK. The results suggested the role of electric acupuncture on the type of fever may associate with the changes of the concentrations of these central substances.
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Thirty albino New Zealand rabbits were divided into three groups at random, ten in each group, which were N~6-2'-O-dibutyryladenosine-3', 5'-cyclic monophosphate (db-cAMP) + taurine (Tau), normal saline (NS) + Tau and db-cAMP+ NS. Tau, dbcAMP and NS were injected into the rabbit's ventricles by intracerebroventricular (ICV) rotate. The results showed that ICV Tau could inhibite the fever induced by ICV dbcAMP in the single long phase hyperthermic duration. The mean thermal response indiexs of the groups within six hours (TRI6) were 7.10?2.447 0.21?1.58 and 12.47?4.60 (P
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PIP: The role of cyclic AMP (cAMP) in the mechanism of abortifacient effect of sulprostone was studied in women with pregnancy of 2-3 weeks of gestation (group 1) or 4-5 weeks of gestation (group 2). Pregnancy was confirmed by determining the beta-subunit of chorionic gonadotropin. The patients received intramuscular injection of sulprostone at 0.5 mg, 2 times with an interval of 4 hours (group 1), or 3 times with an interval of 3 hours (group 2). The cAMP level was determined in the decidual tissue removed immediately after abortion. Sulprostone-induced abortion resulted in marked decrease in the cAMP levels in the decidual tissue (from 491.5+/-70.4 picamol per 1 mg of protein to 224+/55.6 in group 1 and from 377.7+/-55.9 to 95.7+/-18.4 in group 2). Decrease in cAMP level was also observed during spontaneous abortion (122.3+/-28.5, compared with 691.1+/-110.5 during surgical abortion). To determine whether the decrease in cAMP level was associated with direct action of sulprostone, the decidual tissue was incubated with PGE1 in vitro (10, 25 or 50 microg/ml for 15 min). In vitro addition of PGE1 resulted in marked increase in the cAMP level. These findings indicated that decrease in cAMP level during sulprostone-induced abortion was associated not with its direct action on the decidual tissue but rather with reduced blood supply of the decidual tissue caused by uterine contractions.^ieng
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Abortion, Induced , Cyclic AMP/metabolism , Decidua/metabolism , Dinoprostone/analogs & derivatives , Cyclic AMP/deficiency , Female , Humans , Pregnancy , Pregnancy Trimester, FirstABSTRACT
Recent reports on the relationship between caffeine consumption by pregnant women and their infants' risk of miscarriage, low birth weight, preterm delivery and congenital malformations were reviewed. The evidence continues to support the view that moderate consumption of caffeine by pregnant women does not adversely affect their fetuses.
PIP: Reversing the recommendations made by the U.S. Food and Drug Administration of 1981, a review of human epidemiological studies finds no adverse effect of caffeine on pregnancy outcome. Several large studies are discussed on each of the following topics: congenital malformations, miscarriage, low birth weight and preterm delivery. The previous warning was based on gavage feeding of large doses of caffeine to rats, and found a particularly high incidence of facial cleft. Congenital malformations were examined in studies of 12,000 women at Harvard (coffee and tea only), and smaller studies at Boston University, Finland and Tohoku University in Japan (only 0.5% of Japanese women are heavy coffee drinkers, and tea was not considered). A 2nd group of analyses examined low birth weight, at Loma Linda, Harvard, Ottawa, Seattle and elsewhere. Only 1 study, which controlled for nicotine but not education, weight or alcohol, found significantly lower birth weight in 12 women who consumed 300 mg caffeine daily. Preterm birth was not increased in 2 studies. Spontaneous abortion was more likely in 2 studies, the Japanese study with limitations noted above, and the Yale study, which found a higher probability of 2nd trimester miscarriage for both caffeine and alcohol drinkers. These studies apparently prompted the FDA to issue a new statement clearing caffeine, as currently used in foods, of health risk.
Subject(s)
Caffeine/adverse effects , Pregnancy Complications/chemically induced , Pregnancy Outcome/chemically induced , Abnormalities, Drug-Induced/etiology , Abortion, Spontaneous/etiology , Female , Humans , Infant, Low Birth Weight , Infant, Newborn , Infant, Premature , Pregnancy , Risk FactorsABSTRACT
Hemodynamic and clinical observations were performed on 15 patients with severe congestive heart failure treated with Chinese-made milrinone.It was found that after treatment with milrinone,cardiac index was significantly increased,pulmonary capillary wedge pressure,systemic vascular resistance,and pulmonary vascular resistance were significantly decreased,and heart rate and myocardialoxygen consumption remained unchanged.Tolerance exercise test was prolonged and plasma cyclic AMP was increased.No thrombocytopenia or pro-arrhythmic effect was observed.The total clinical efficacy of the drug was 86.7%.These facts suggest that the Chinese-made milrinone is the durg of choice for the management of patients with congestive heart failure.
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Human spermatozoa were incubated with gossypol acetic acid (100 micrograms/1 X 10(6) spermatozoa/ml) at 37 degrees C for 30 min. The drug treatment inhibited the spermatozoal motility significantly. Washing of the spermatozoa, after gossypol treatment, did not effect their motility. A partial revival in the motility of the spermatozoa was observed when gossypol treated spermatozoa were incubated, after washing, with motility stimulating factors, e.g. theophylline, dibutyryl-cAMP and Kallikrein.
PIP: This brief experiment asked the question whether the inhibition of sperm motility imposed by gossypol is reversible by washing away the polyphenol. Human spermatozoa were incubated in Bakers buffer, 1 million/ml at 37 degrees C. for 30 minutes in the presence of 100 mc gossypol. Gossypol was dissolved in ethanol and diluted to 0.01% with distilled water, then 0.01 ml was added to 1.0 ml spermatozoa. Controls contained 0.1 ml of 0.01% ethyl acetate. Gossypol reduced motility from a range of 66.1 to 79.4% to a range of 32.1 to 38.1% (p0.01). Washing the spermatozoa by centrifuging twice at 343 x g for 10 min. and reincubating in a buffer had no effect on motility. Addition of the sperm motility stimulating agents theophylline 10 mM, kallikrein 100 KIU or dibutyryl-caMP after washing away the gossypol restored motility to 55.1%, 58.7% and 50.0%, respectively (p0.01).
Subject(s)
Gossypol/analogs & derivatives , Sperm Motility/drug effects , Bucladesine/pharmacology , Gossypol/pharmacology , Humans , In Vitro Techniques , Kallikreins/pharmacology , Male , Theophylline/pharmacologyABSTRACT
AIM: To observe the changes of AC activity and content of cAMP at different time point in hypothalamus of rats with fever and hypothermia. METHODS: Radioisotope method was used to measure the enzymatic activity of AC and the content of cAMP. RESULTS:(1)The fresh yeast caused fever after making model 4 h( P
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AIM: To determine whether vasorelaxation induced by cAMP is dependent on eNOS activation.METHODS: A constant perfusion model of rabbit femoral artery was used to observe the changes of perfusion pressure before and after intra-arterial infusing different concentrations of dbcAMP(a membrane-permeable cAMP analogue) or forskolin(a direct stimulator of adenyl cyclase) and the effects of NOS inhibitor L-NAME on their actions.RESULTS: Both dbcAMP and forskolin induced concentration-dependent relaxation of rabbit femoral artery bed and NOS inhibitor L-NAME partly inhibited this vasorelaxation.CONCLUSION: Vasorelaxation induced by cAMP is mediated partly through eNOS activation.
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AIM: To investigate the mechanism of Qing-Kai-Ling(QKL) Injecta suppressing endotoxin(ET)-induced fever in rabbits.METHODS: ①The ET-induced fever model was established in rabbits. QKL injecta was administered intravenously, and the febrile response of rabbits was observed. The cAMP content in the hypothalamus (HP) and arginine vasopressin (AVP) content in the ventral septal area (VSA) were determined by radioimmunoassay.RESULTS: ①The maximal increment in body temperature (△T)[(1 68?0 46)℃],6 h thermal response index (TRI 6)(29 59?10 39), cAMP content in the HP[(2 90?0 40) nmol/g] and AVP content in the VSA[(47 32?3 77) ng/g] in the ET group were significantly higher than those in the normal saline (NS) group[(0 08?0 11)℃,-0.15?4 29,(1 05?0 37) nmol/g,(11 78?7 87) ng/g, respectively] and the QKL+ET group[(0 48?0 18)℃, (13.71?3.29), (1 37?0 23) nmol/g,(25 24?2 61) ng/g, respectively].②The positive correlation was observed between the cAMP content in the HP and the fluctuation of body temperature ( r=0.904,P
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AIM and METHOD: Human endothelial nitric oxide synthase (eNOS) gene was transfected into human phagocytic cell U937 and the effects of gene transfer on cytokines and cAMP production were observed. RESULTS: A functional eNOS was stably expressed in transfected U937 cells, but NO release was undetectable in intact transfectants. However, eNOS gene expression upregulated tumor necrosis factor - a release and downregulated interleukin - 10 and cAMP production in either presence or absence of NOS inhibitor N? - monomethyl - L - arginine. CONCLUSION: The function of tranfected eNOS gene product showed cellular speciality. The effector molecule that changed the produced pattern of cytokines and cAMP in phagocytic cells seems not likely the nitric oxide.
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The ?-adrenoceptor densities and cAMP contents of the lung tissues ofthe experimental allergic asthmatic guinea pigs with and without treatment of galvano-acupuncture were measured by radioligand binding and protein binding assays. The relaxreaction of lung and tracheal strips toward isoproterenal were measused by pharmacological assay. The results were as follows: (1) The ?-receptors densities and cAMP contentsin lung tissues of asthmatic guinea pigs were both lower than those of the normal controls(P
