Objective: To establish the fingerprint of the extract from the rhizome of Alpinia officinarum, to investigate the effects on the proliferation, tyrosinase activity, and melanogenesis in melanoma B16 cells, and to analyze the spectrum-effect relasionship. Methods: The fingerprint of alcohol extract from the rhizome of A. officinarum (G1), water sediments of G1 (G2), macroporous resin purified extract of G2 (G3), and refined polyamide extract of G3 (G4) were established by HPLC, and the proliferation of each extract on B16 cells was measured by MTT. The tyrosinase activity and melanin content were measured by colorimetry assay. The spectrum-effect relasionship was analyzed by grey relation analysis. Results: Compared with the control group, the proliferation of B16 cells was promoted significantly after treatment with 1.00-5.00, 0.50-5.00, 0.25-5.00, and 0.25-10.00 μmol/L of G1, G2, G3, and G4, respectively. The tyrosinase activity was promoted by G1 and G2 with 0.50 μmol/L, and G3 and G4 with 0.50-1.00 μmol/L (P < 0.05, 0.01); The synthesis of melanin was promoted after the treatment with 0.5 and 5.00 μmol/L of G1, 0.50-5.00 μmol/L of G2 and G4, and 0.25-5.00 μmol/L of G3 (P < 0.01); At the same concentration, G4 had the strongest effect to promote the cell proliferation and to increase the tyrosinase activity. The galangin (peak 7) had the highest correlation with pharmacodynamics, and the contents of galangin in G1, G2, G3, and G4 were 27.61, 88.72, 348.53, and 693.35 mg/g, respectively. The proliferation, tyrosinase activity, and galangin content were promoted significantly after the treatment with galangin (0.25-10.00 μmol/L) by the verification experiment. Conclusion: The effect of the extract from the rhizome of A. officinarum on the proliferation, tyrosinase activity, and melanogenesis of B16 melanoma cells could be enhanced with the increased content of galangin, which demonstrates that the galangin is one of the efficient substances in the rhizome of A. officinarum for the treatment of vitiligo.

Objective To investigate the effects of melatonin (MT) on rats with acute paraquat (PQ) poisoning. Method Fifty-four Sprague-Dawley (SD) rats were randomly divided into three groups (each group 18 rats) and given the following treatment: intragastric injection of PQ at 50 mg/kg (PQ); intragastric injection of paraquat followed by intraperitoneal injection of MT at 10mg/kg once a day (MT); intragastric injection of normal saline (Control). Serum assays for malondialdehyde (MDA) levels and superoxide dismutase (SOD) and glu tathione peroxidase (GSH-Px) activities were determined on the 1st, 3rd and 7th day post treatment. Clinical manifestations of poisoning and pathological changes in the lungs were also observed. Results Serum MDA levels were significantly increased (P ＜ 0.01), and SOD and GSH-Px activities significantly decreased (P ＜ 0.05) in the PQ group compared to the control group. Serum MDA levels were significantly decreased, and serum SOD and GSH-Px activities increased in MT group compared to the PQ group (P ＜ 0.05). Clinical manifestations of intoxication and pathological lung changes were also ameliorated in poisoned rats treated with MT. Condutions Administration of MT alleviates clinical manifestations of acute paraquat poisoning in rats by Limiting the damage from lipid peroxidation.

Animals ; Humans ; Hypertension, Pulmonary ; etiology ; Inflammation Mediators ; physiology ; Neutrophil Activation ; Platelet Activating Factor ; antagonists & inhibitors ; physiology ; Pulmonary Edema ; etiology ; Respiratory Distress Syndrome, Adult ; drug therapy ; etiology

Objective To investigate the effect of recombinant human bone morphogenetic protein 2 (rhBMP-2) on the osteogenic activities of human osteosarcoma cell line SaOS-2. Methods SaOS-2 cells were exposed to rhBMP-2 for 12,24,48 h at 0(control) ,2,20,200 μg/L, respectively. The mRNA expression of alkaline phosphatase(ALP) and bone gla(BCP) were detected by real time polymerase chain reaction. Results The mRNA expression of ALP and BGP of SaOS-2 cells increased gradually with rhBMP-2. The mRNA expression of ALP of the 20 μg/L group exposed for 48 h(1.60 ± 0.64), and the 200 μg/L group exposed for 12,48 h(1.70 ± 0.41, 1.80±0.19) were significantly higher than those of control (12 h: 0.80±0.25, 48 h: 0.74±0.21, allP<0.05). The mRNA expression of BGP of the 2 μg/L group exposed for 24 h(1.67 ± 0.33), the 20 μg/L group exposed for 12,24 h(2.42 ± 0.13,1.82 ± 0.14) and the 200 μg/L group exposed for 12,24 h(1.46 ± 0.11,1.24 ± 0.07) were significantly higher than those of control( 12 h: 1.01 ± 0.14, 24 h: 0.84 ± 0.12, all P< 0.05). Conclusions rhBMP-2 can promote the mRNA expression of ALP and BGP of SaOS-2 cells. They have a dose-response relationship, but represent a different dose-response effect.

Objective To observe the effects of recombinant human parathyroid hormone 1 to 34(referred to as hPTH) on the expression level of alkaline phosphatase(ALP) and bone gla protein(BCP) in human osteosarcoma cell line SaOS-2(referred to as SaOS-2 cells). Methods SaOS-2 cells were subcultured and treated with 1, 10 and 100 nmol/L hPTH for 12, 24 and 48 h. Total cellular RNA was extracted, cDNA was synthesized by reverse doses of hPTH, different duration of action, and their interaction on the expression level of ALP mRNA of SaOS-2 cells was significantly different(F = 29.32, 2.92, 7.64, all P < 0.05). The expression level of ALP mRNA(0.78 ± 0.43, 0.71 ± 0.05, 0.75 ± 0.19, 0.76 ± 0.14) of SaOS-2 cells after treatment with 0, 1, 10 and 100 nmol/L hPTH for 48 h was lower than those of treated for 12 h(1.01 ± 0.16, 1.37 ± 0.38, 1.49 ± 0.16, 2.52 ± 0.70, all P< 0.05) and 24 h (1.80 ± 0.47, 1.30 ± 0.36, 1.27 ± 0.17, 1.17 ± 0.11, all P< 0.05). The expression level of ALP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 12 hours was higher than that of the control(P < 0.05); the expression level of ALP mRNA of SaOS-2 cells after treatment with 1, 10 and 100 nmol/L hPTH for 24 h interaction on the expression level of BGP mRNA of SaOS-2 were significantly different (F = 8.26, 10.33, 5.51, all P< 0.05). The expression level of BGP mRNA(1.17 ± 0.28, 0.98 ± 0.08, 0.92 ± 0.17 and 0.84 ± 0.59) of SaOS2 cells after treatment with 0, 1, 10 and 100 nmol/L hPTH for 48 h was lower than those of treated for 12 h( 1.01 ± 0.14, 1.21 ± 0.18, 1.34 ± 0.30, 1.68 ± 0.62, all P< 0.05), and 24 h(1.71 ± 0.35, 1.41 ± 0.47, 1.28 ± 0.31 and 1.01 ± 0.18, all P < 0.05). The expression level of BGP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 12 h was higher than that of those groups treated with 0 and 1 nmol/L hPTH(all P< 0.05). The expression level of BGP mRNA of SaOS-2 cells after treatment with 10 and 100 nmol/L hPTH for 24 h and 48 h was lower than those of the control(all P < 0.05). The expression level of BGP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 24 hours was lower than that the group treated with 1 nmol/L hPTH(P < 0.05). Conclusions In vitro, hPTH significantly enhances osteogenic activities of human osteoblast in a short time, however, with prolonged stimulation time, osteogenic activity can show a downward trend.

Objective To explore whether extracellular signal-reg-ulated kinase (ERK) signaling pathway is involved in the hypoxia-inducible factor-1α (HIF-1α) expression in the hippocampus of epileptic rats.Methods A total of 208 21-d old SD rats were equally randomized into status epilepticusgroup (SE,n=96),normal control group (NS,n=96) and PD98059 (the ERKsignaling pathway specific inhibitor) treatment group (n=16),respectively; SE rat models of the SE group were induced by intraperitoneal injection of 1％ PTZ (80 mg/kg),and rats of the NS group received injection of normal saline (NS); 0.5,1,1.5,6,12 and 24 h after the inducement,the mRNA and protein expressions of HIF-lα and ERK1/2 in the hippocampus of rats in these two groups were examined by RT-PCR and Westem blotting.For rats in the PD98059 treatment group,PD98059 was intraperitoneally injected 10 minutes before intraperitoneal injection of pentylenetetrazol; 1 h after the inducement,the mRNA expressions of HIF-1α and ERK1/2 in the hippocampus of rats were examined by RT-PCR,while 1.5 h after the inducement,the protein expressions of HIF-1α and ERK1/2 in the hippocampus of rats were examined by Western blotting.The results of the PD98059 treatment group would be compared with those of the SE group.Results Compared with the NS group,the mRNA and protein expressions of HIF-1α and ERK 1/2 in the hippocampus of rats in the SE group after SE increased significantly; the peakexpression time ofERK1/2 mRNA was 1 h after SE (1.112±0.126 h),and the ERK1/2 protein mostly expressed at 1.5 h after SE (1.127±0.155 h).As to HIF-1α mRNA and its protein,the peak expression time was 1.5 h after SE (0.589±0.090 h) and 6 h after SE (0.230±0.052 h),respectively (P＜0.05).Compared with the SE group,the mRNA and protein expressions of HIF-1α and ERK1/2 in the hippocampus of all the rats in the PD98059 treatment group after SE decreased significantly (P＜0.05);positive correlation between HIF-1α and ERK1/2 mRNA in the SE group was noted (r=0.688,P=0.000).Conclusion ERK signaling pathway is activated in the epileptic rats and it participates in the expressionof HIF-1α in the hippocampus.

OBJECTIVETo study on the effect of acteoside on learning and memory of dementia mice.

METHODMice were orally administered with acteoside for 10 days. Scopolamine was used to establish the acquired learning disability in mice. Their learning and memory were detected with a behavioral experiment (step-down test). After the behavior test, corticocerebral and hippocampus tissues of mice were detected with biochemical indexes, including GSH-Px, T-SOD, MDA, TChE and contents of protein in brain tissues.

RESULTMice were administered with acteoside for 10 d in advance to alleviate the acquired learning disability induced by scopolamine. Compared with the model group, acteoside increased the latency period in the step-down test and reduced error times. Besides, acteoside increased the activity of GSH-Px, T-SOD, TChE and protein content in their brain tissues, but decreased MDA content.

CONCLUSIONActeoside can significantly alleviate the acquired learning disability in mice induced by scopolamine. Its mechanism may be related with its effect of inhibiting the generation of free radicals in mice and improving the function of the central cholinergic system.

Animals ; Behavior, Animal ; drug effects ; Brain ; drug effects ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Glucosides ; administration & dosage ; pharmacology ; Glutathione Peroxidase ; metabolism ; Learning ; drug effects ; Male ; Memory Disorders ; chemically induced ; drug therapy ; Mice ; Phenols ; administration & dosage ; pharmacology ; Scopolamine Hydrobromide ; adverse effects ; Superoxide Dismutase ; metabolism

OBJECTIVETo establish a HPLC method for determining acetoside in rat plasma and to investigate the pharmacokinetic characteristics of acetoside in rats.

METHODSix rats were orally administered with 150 mg x kg(-1) acetoside and their blood samples were collected at different time points. The plasma concentration of acetoside was determined by reserved HPLC, and the pharmacokinetic parameters were calculated by DAS 2.0 software.

RESULTThe regression equation of acetoside in rats plasma was Y = 3.509 8X-0.096 8 (r = 0.996 8), which showed a good linear relation at 0.125-2.5 mg x L(-1). The method showed a recovery of more than 85%, and both inter-day and intra-day RSDs were less than 15%. After the oral administration of 150 mg x kg(-1) acetoside, the concentration-time curves of acetoside were expressed in a open two-compartment model. The main pharmacokinetics parameters of T(max), C(max), t(1/2alpha), t(1/2beta), AUC(0-t), AUC(0-infinity), CL/F, V/F and K(a) were respectively 0.36 h, 1.126 mg x L(-1), 0.759, 4.842 h, 3.134, 3.766 mg x h x L(-), 87.089 L x h(t) x kg(-1), 207.704 L x kg(-1) and 6.345 h(-1) respectively.

CONCLUSIONIt is first time to establish such a HPLC method to determine the concentration of acetoside in plasma. The method is so highly specified and sensitive that it can ble used in quantitative analysis in vivo on acetoside.

Animals ; Chromatography, High Pressure Liquid ; Female ; Glucosides ; chemistry ; pharmacokinetics ; Male ; Phenols ; chemistry ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate NF-kappaB activity and the expression of phosphorylated p38 MAPK protein in lung tissue of acute paraquat poisoned rats and the effect of MT.

METHODSOne hundred and twenty-eight Sprague-Dawley (SD) rats were randomly divided into three experimental groups: poisoned group, MT group and control group. On the 1st, the 3rd, the 7th and the 14th day after exposure, levels of malondialdehyde (MDA) in serum were detected, NF-kappaB activity in the lung tissues was assessed by electrophoresis mobility shift assay (EMSA), the expression of the phosphorylated p38 MAPK was evaluated by Western blot method, the lung pathological changes of rats were observed.

RESULTSThe level of malondialdehyde (MDA) in serum increased significantly in poisoned group on the 1st day (4.45 +/- 1.23), the 3rd day (3.77 +/- 1.12) and the 7th day (2.84 +/- 0.96) nmol/ml compared with that in control group (1.36 +/- 0.52) nmol/ml (P < 0.01). There was a significant decrease in MT group on the 1st day (2.68 +/- 0.85), the 3rd day (1.97 +/- 0.74) and the 7th day (1.53 +/- 0.62) nmol/ml compared with poisoned group (P < 0.05). The expression of the phosphorylated p38 MAPK and NF-kappaB activity in lung tissue of poisoned group significantly increased compared with control group (P < 0.01). There was a significant decrease in NF-kappaB activity and expression of the phosphorylated p38 MAPK in the lung tissues in MT group compared with poisoned group (P < 0.05).

CONCLUSIONNF-kappaB and p38 MAPK could play an important role in lung injury of poisoned rats. MT may inhibit the expression of NF-kappaB and phosphorylated p38 MAPK, and therefore might have the therapeutical effect on acute paraquat poisoning.

Acute Lung Injury ; chemically induced ; metabolism ; pathology ; Animals ; Disease Models, Animal ; Female ; Lung ; drug effects ; metabolism ; pathology ; Male ; NF-kappa B ; metabolism ; Paraquat ; poisoning ; Phosphorylation ; drug effects ; Rats ; Rats, Sprague-Dawley ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo investigate the effect of acteoside on SK-N-SH nerve cell injury induced by okadaic acid (OA).

METHODSK-N-SH nerve cells were processed with 20 nmol * L OA to establish the Alzheimer's disease (AD) cellular model, and 5, 10, 20 mg . L-1 acteoside was used to antagonize against its effect. Cell morphology was observed under inverted microscope. The cell survival rate was detected with MTT, and the LDH release rate was measured by enzyme label kit. Western blot was applied to determine the expression of phosphorylation tau proteins in nerve cells.

RESULTThe acteoside could significantly improve SK-N-SH cell morphology, enhance the cell survival rate, decrease the cell LDH release rate and the expression of phosphorylated tau proteins at p-Ser 199/202 and p-Ser 404 sites, up-regulated the expression of at non-phosphorylated tau proteins at Ser 202 site and Ser 404 sites.

CONCLUSIONActeoside has significant protective effect on nerve cell injury induced by OA.

Alzheimer Disease ; metabolism ; Cell Line ; Cell Survival ; drug effects ; Glucosides ; pharmacology ; Humans ; Okadaic Acid ; Phenols ; pharmacology ; tau Proteins ; metabolism