Elucidation of the FKBP25-60S Ribosomal Protein L7a Stress Response Signaling During Ischemic Injury.

BACKGROUND/AIMS: Peptidyl-prolyl cis-trans isomerase FKBP25 is a member of the FK506-binding proteins family which has peptidyl-prolyl cis/trans isomerase domain. The biological function and pathophysiologic role of FKBP25 remain elusive. METHODS: The spatio-temporal changes in expression of endothelial FKBP25 upon oxygen-glucose deprivation (OGD) treatment were examined by Western blot and immunofluorescence. The immunoprecipitation and fluorescence resonance energy transfer (FRET) were used to address the interacting proteins with FKBP25. RESULTS: In the present study, nuclear translocation of FKBP25 was observed following OGD in cultured endothelial cells. Intriguingly, FKBP25 nuclear translocation was further validated in peroxynitrite (ONOO-)-treated endothelial cells. Coimmunoprecipitation and FRET data indicated that FKBP25 translocated into the nucleus, in which it interacted with 60S ribosomal protein L7a, while overexpression FKBP25 protect endothelial cells against OGD injury. CONCLUSION: Our findings reveal that the nuclear import of FKBP25 and binding with 60S ribosomal protein L7a are protective stress responses to ischemia/nitrosaive stress injury.

[Hypouricemic effect of ethanol extracts from Dioscoreae Nipponicae Rhizoma].

OBJECTIVE: To investigate the effect of ethanol extracts from Dioscoreae Nipponicae Rhizoma on hyperuricemic mice. METHODS: The hyperuricemia was induced by gavage of hypoxanthine and subcutaneous injection of potassium oxonate (model A) or subcutaneous injection of uric acid (model B) in ICR male mice. The mice in ethanol extracts groups were administrated with Dioscoreae Nipponicae Rhizoma ethanol extracts 5.4 g/kg by gavage, the positive control groups were given with 10 mg/ml allopurinol or 5 mg/ml benzbromarone by gavage, respectively. The plasma uric acid levels were measured by using HPLC. RESULTS: The plasma uric acid levels of model group, control group and ethanol extract group in model A mice were (40.03±27.24), (4.08±1.47) and (18.10±8.87) g/mL (compared with model group, P <0.05), respectively. The plasma uric acid levels of model group, control group and ethanol extract group in model B mice were (18.57±3.83), (4.29±2.36) and (15.36±2.71) g/mL (compared with model group, P <0.05), respectively. CONCLUSION: The ethanol extracts from Dioscoreae Nipponicae Rhizoma have certain hypouricemic effect in hyperuricemic mice induced by hypoxanthine and potassium oxonate or by uric acid.

[Extract of Herba Siegesbeckiae on mouse rheumatoid arthritis induced by anthrogen-CIA monoclonal antibody].

OBJECTIVE: To investigate the anti-inflammatory effect of Herba Siegesbeckiae extracts on mouse rheumatoid arthritis induced by arthrogen-CIA monoclonal antibody. METHODS: The rheumatoid arthritis was induced by arthrogen-CIA arthritogenic monoclonal antibody in mice. The sandwich enzyme-linked immunosorbent assay was used to determine the concentration of IL-1ßin mouse serum,and the content of IL-6,IL-17 and MMP-3 in supernatant of tissue homogenate of hind limb below the stifle of mice. One-way ANOVA was used for data analysis. RESULTS: The toe swelling was attenuated in Siegesbeckiae group than that in model group [(0.218 ± 0.0307)cm(3) compared with (0.2545 ± 0.0179)cm(3), P<0.05]. The serum IL-1ß level in Siegesbeckiae group was lower than that in model group [(63.74 ± 21.74)pg/ml compared with (104.96 ± 31.22)pg/ml, P<0.01]. The contents of IL-6, IL-17 and MMP-3 in tissue supernatants of Siegesbeckiae group were all lower than those of model group [(171.10 ± 48.35)pg/ml compared with (249.64 ± 75.08)pg/ml, P<0.05; (115.42 ± 56.52)pg/ml compared with (208.40 ± 88.54)pg/ml, P<0.05;(3660.31 ± 1680.99) pg/ml compared with (5420.79 ± 1201.43)pg/ml, P<0.05, respectively]. CONCLUSION: The extract of Herba Siegesbeckiae has anti-inflammatory effect on mouse rheumatoid arthritis induced by mixed arthrogen monoclonal antibody.

[Effects of Corbrin Shugan capsule on dimethylnitrosamine-induced hepatic fibrosis in rats].

OBJECTIVE: To investigate the effects of Corbrin Shugan capsule on dimethylnitrosamine (DMN)-induced hepatic fibrosis in rats. METHODS: Hepatic fibrosis was induced by DMN in AD rats. The serum concentrations of III pro-collagen (III PC),laminin (LN) and tissue inhibitor of metalloproteinase-1(TIMP-1) were determined with ELISA. The concentration of albumin (ALB) in sera and the content of hydroxyproline (Hyp) in liver tissues were determined with chemical colorimetric and HPLC, respectively. The fibrosis area was measured with Motic Med 6.0 digital medical image analysis system. RESULTS: Compared to model group the high-dose (450 mg kg(-1)),mid-dose (270 mg kg(-1)) and low-dose (90 mg kg(-1)) groups of Corbrin Shugan capsule had significantly lower serum content of III PC [34.46 ± 13.95),(36.15 ± 9.46), and (40.58 ± 7.72)ng ml(-1) compared with (49.38 ± 10.95)ng ml(-1),P<0.05 or P<0.01],TIMP-1 [(16.65 ± 4.24),(16.66 ± 4.34),and (18.99 ± 6.05)ng ml(-1) compared with (30.84 ± 14.48)ng ml(-1), P<0.05 or P<0.01], LN [(12.94 ± 4.29), (12.96 ± 3.21),and (15.32 ± 8.00)ng ml(-1) compared with (30.22 ± 17.00)ng ml(-1),P<0.05 or P<0.01] and smaller hepatic fibrosis area [(0.02240 ± 0.01337), (0.02176 ± 0.01460) and (0.02384 ± 0.01405)µm(2) compared with vs (0.03929 ± 0.01732)µm2, P<0.05 or P<0.01]; the high-dose and mid-dose groups of Corbrin Shugan capsule had significantly lower content of Hyp in liver tissues [(0.77 ± 0.09) and (0.81 ± 0.09)µg µmg(-1) compared with (1.06 ± 0.33)µg mg(-1),P<0.05 or P<0.01]; and the high-dose group of Corbrin Shugan capsule significantly increased the content of ALB in sera [(34.02 ± 4.17)g L(-1) compared with (30.25 ± 4.21)g L(-1),P<0.05]. CONCLUSION: Corbrin Shugan capsule is effective in treatment of DMN-induced hepatic fibrosis in rats.

[Corbrin shugan capsule for treatment of alcoholic hepatic fibrosis in rats].

OBJECTIVE: To investigate the therapeutic effect of Corbrin shugan capsule for treatment of alcoholic hepatic fibrosis in rats. METHODS: The rat model of alcoholic hepatic fibrosis was induced by intragastric administration of alcohol repeatedly. The serum procollagen III (PC III), laminin (LN) and tissue inhibitors of metalloproteinase-1 (TIMP-1) levels were measured with ELISA, and the content of hydroxyproline (Hyp) in liver tissue were determined with colorimetric method. Collagen deposition in liver tissue was observed with Masson's staining, and the fibrosis area was measured with digital medical image analysis system (Motic Med 6.0). RESULTS: Compared with the model control group, the serum TIMP-1 and LN levels and hepatic fibrosis area in liver tissue significantly decreased in Corbrin shugan capsule groups with doses of 0.09,0.27 and 0.45 g*kg(-1), and the serum PC III and the Hyp contents in liver tissue also decreased of Corbrin shugan capsule groups with doses of 0.27 and 0.45g*kg(-1). CONCLUSION: Corbrin shugan capsule can decrease serum PC III, TIMP-1 and LN levels and Hyp levels in liver tissue and hepatic fibrosis area in rats, indicating it may have therapeutic effect on alcoholic hepatic fibrosis.

[Determination of hydroxyproline in rat kidney tissue by HPLC with pre-column derivatization].

OBJECTIVE: To establish a pre-column derivatization HPLC method for quantitation of hydroxyproline (Hyp) in rat kidney tissue by reversed-phase assay. METHODS: Rat kidney samples were hydrolyzed in 6 mol x L⁻¹ HCl under 110 degree for 24 h, then 9-fluorenylmethyl chloroformate (FMOC) was added for the pre-column reaction. The HPLC analysis was performed on a Phenomenex C18 column using sodium acetate buffer, methanol and acetonitrile as mobile phase with a flow rate of 1.0 ml min⁻¹ and UV detective wavelength 265 nm at 40 degree. RESULTS: Linear range was 15.30-612.00 mg x L⁻¹ (correlation coefficient was 0.9999). Recovery rate was 97.4%-103.9%. CONCLUSION: The established method is simple, accurate and sensitive to analyze Hyp content in rat kidney tissue.

[Protective effect of Jiangbaiweiyan tablet on ethanol-induced gastric mucosa injury in rats].

OBJECTIVE: To investigate the effect of Jiangbaiweiyan tablet, a Chinese medicine compound composed of Alpinta Officinarum, Cyperus Rotundus, Bulbus Lilii and Rlindera Strychnifolia, on ethanol-induced gastric mucosa injury in rats. METHODS: Acute gastric ulcer was induced in rats with absolute ethyl alcohol. The ulcer index was used to evaluate the extent of the gastric mucosa injury. RESULTS: The ulcer indexes of the model group, the mid-dose (1.08 g x kg(-1) x d-(-1)1) and high-dose (2.16 g x kg(-1) x d-(-1)) of Jiangbaiweiyan tablet groups were 141.58±47.43, 24.83±23.04 and 2.12±2.58, respectively (P<0.01). CONCLUSION: Jiangbaiweiyan tablet has protective effects on ethanol-induced gastric mucosa injury in rats, which may be related to anti-oxidation and enhancing tissue regeneration capacity.

[Drug permeability through buccal mucosa].

OBJECTIVE: To investigate the effects of physicochemical properties of drug on its permeability through buccal mucosa. METHODS: Benzoic acid, caffeine and hydrocortisone were selected as model drugs and their permeability coefficients through isolated porcine buccal mucosa were determined. RESULT: The permeability coefficients of benzoic acid, caffeine and hydrocortisone through isolated porcine buccal mucosa were (1.01 x 10(-4)+/-1.64 x 10(-5))cm.s(-1)ì(2.75 x 10(-5)+/-1.79 x 10(-6))cm.s(-1) and (2.49 x 10(-5)+/-6.32 x 10(-6))cm.s(-1), which were 14,989 and 389 times as those through human skin, respectively. CONCLUSION: The permeability of drugs through buccal mucosa seems to be much better than that through human skin.

Predicting blood-brain barrier penetration from molecular weight and number of polar atoms.

A simple three-descriptor model to predict blood-brain barrier is derived from a training set of 78 compounds: logBB=-9.880 x 10(-6)M(W)(2)+7.339 x 10(-3)M(W)-0.2268n(pol)-0.1143 (n=78, r(2)=0.74), where logBB is the logarithm of the ratio of the steady-state concentration of the compound in the brain to concentration in the blood, M(W) is the molecular weight, n(pol) is the number of polar atoms (oxygen, nitrogen, and attached hydrogen), n is the number of compounds, and r is the correlation coefficient. The model is validated through use of leave-one-out procedure and an external test set (25 compounds). The model is suitable for the rapid prediction of the blood-brain barrier penetration of drug candidates because of its predictive ability and simplicity.

[Synthesis, spectral analysis, antibacterial and antitumor activities of Co(II) and Ni(II) ofloxacin complexes].

The interactions of cobalt sulphate and nickel acetate with ofloxacin (ofo, I), a 4-quinoline derivative, were studied. The hydrothermal technique was adopted in this work. The isolated solid complexes were characterized by elemental analysis, infrared spectra, electronic spectra, fluorescence spectra and thermal analysis. The results support the formation of complexes of the formula Co(ofo)2 x 4H2O (II) and Ni(ofo)2 x 4H2O (III). The infrared spectra of the isolated solid complexes suggested that ofo acts as bidentate ligands through one of the oxygen atoms of the carboxylic group and the ring carbonyl oxygen atom. Thermogravimetric (TG) and its differential (DTG) were carried out for the complexes. The data obtained indicated that the thermal decomposition of the two complexes in inert atmosphere proceeded approximately with two main degradation steps. I showed a intense fluorescence in solid state at lamda(ex)/lamda(em) = 365 nm/461 nm, and two complexes displayed weakly similar emission maximum at 470 nm in the powder samples at ambient temperature, while the emission of II and III may be mainly originated from the intraligand excited states of I. I, II and III were assayed against two kinds of gram-positive and two kinds of gram-negative bacteria by in vitro doubling dilutions method. The results indicated that II and III have the similar minimal inhibitory concentration (MIC) as the I against S. Aureus, M. Lutens, E. Coli and P. Aeruginosa. The inhibitory effect of I, II and III on leukemia HL-60 cell line has been measured by using MTT (Methyl-Thiazol-Tetrozolium) method and that on liver cancer BEL-7402 cell line measured by SRB (Sulphurhodamin B) method. The results indicated that the complexes in the high concentration have inhibitory effect on HL-60 cell line.

[Prediction of plasma protein binding of cephalosporins from polar molecular surface areas].

OBJECTIVE: To predict the plasma protein binding rate of cephalosporins from their molecular structural parameters. METHODS: The minimum energy conformations of cephalosporins were obtained from the optimization of the standard molecular geometry with the semiempirical self-consistent field molecular orbital calculation AM1 method; Mont Carlo method was used to calculate the polar molecular surface areas; the stepwise multiple regression analysis was used to obtain the correlation equations. RESULTS: The plasma protein binding rate of cephalosporins (fb) was well correlated with their molecular weights (MW) and surface areas of hydrogen-bonding donors (SH). The regression equation was: fb=0.5057+2.861x10(-3) MW-0.1572SH+4.714x10(-3) SH2(n=22, r=0.9042). CONCLUSION: Plasma protein binding of cephalosporins is closely related with their lipophilicity and hydrogen- bonding potential. The plasma protein binding rate of cephalosporins can be predicted from their molecular weights and surface areas of hydrogen-bonding donors.

[Prediction of human intestinal absorption from net polar atomic charges of drug molecules].

OBJECTIVE: To predict human intestinal absorption and permeability coefficients in Caco-2 cell monolayers from net polar atomic charges of drug molecules. METHODS: The net atomic charges and the volumes of drug molecules were obtained with the semiempirical self-consistent field molecular orbital calculation CNDO/2 method and Mont Carlo method respectively, using the minimum energy conformation obtained from the optimization of the standard molecular geometry with the molecular mechanics MM+ method. The stepwise multiple regression analysis was used to obtain the correlation equations. RESULT: Both percent of human intestinal absorption and permeability coefficients in Caco-2 cell monolayers of drug molecules were well correlated with the sum of the net atomic charges of all hydrogen-bonding donors (sigmaQH) and the sum of the net atomic charges of all hydrogen-bonding acceptors (sigmaQN, 0). The more the net positive atomic charges of hydrogen-bonding donors and the net negative atomic charges of hydrogen-bonding acceptors, the less were the percent human intestinal absorption and permeability coefficients in Caco-2 cell monolayers of drug molecules. CONCLUSION: Drug absorption in human intestines is closely related with its hydrogen-bonding potential. The drug molecules with weaker hydrogen-bonding potential have greater percent human intestinal absorption. The net polar atomic charges can be computed simply, so they can be used in high throughput screening of oral drugs.

[Correlation between absorption rates of beta-adrenoreceptor antagonists in rat small intestine and their molecular structures].

OBJECTIVE: To study the correlation between the absorption rate constants of beta-adrenoreceptor antagonists in rat small intestinal segments and their molecular structural parameters. METHODS: The net atomic charges and the molecular volumes of 11 beta-adrenoreceptor antagonists were obtained with the semiempirical self-consistent field molecular orbital calculation CNDO/2 method and Mont Carlo method respectively, using the minimum energy conformation obtained from the optimization of the standard molecular geometry with the molecular mechanics MM+ method. The stepwise multiple regression analysis was used to obtain the correlation equations. RESULTS: The absorption rate constants of beta-adrenoreceptor antagonists in rat jejunum or ileum were well linearly correlated with the sum of the net charges of all hydrogen atoms and the molecular volumes. The beta-adrenoreceptor antagonist with higher lipophilicity, weaker hydrogen-bonding potential,and smaller molecular volume had greater absorption rate constants. CONCLUSION: The absorption rate constants of beta-adrenoreceptor antagonists in rat small intestinal segments are mainly related with their lipophilicity,hydrogen-bonding potential and molecular size.

[Prediction of percutaneous drug permeability using modified theoretical linear solvation energy relationship].

OBJECTIVE: To predict the percutaneous drug permeability coefficients with modified regression equation. METHODS: The semiempirical self-consistent field molecular orbital calculation AM1 method was used to calculate the quantum chemical parameters and the modified theoretical linear solvation energy relationship was used to obtain the regression equation of the permeability coefficients of drugs through human epidermis. RESULT: The permeability coefficients (P) of 36 nonelectrolytes were well linearly correlated with their theoretical descriptors including molecular volume (V), hydrogen bond acidity (sum alpha(2)(H)), hydrogen bond basicity (sum beta(2)(H)) and polarizability index (pi(1)). The regression equation was logP=-6.790+1.571 V+0.1550 pi(1)-1.295 sum alpha(2)(H)-2.485 sum beta(2)(H)(n=36,r=0.9777). CONCLUSION: The modified theoretical linear solvation energy relationship can be used to predict the skin permeability of drugs.