The objective of this study was to enhance the oral bioavailability (BA) of zanamivir (ZMR) by increasing its intestinal permeability using permeation enhancers (PE). Four different classes of PEs (Labrasol(R), sodium cholate, sodium caprate, hydroxypropyl beta-cyclodextrin) were investigated for their ability to enhance the permeation of ZMR across Caco-2 cell monolayers. The flux and Papp of ZMR in the presence of sodium caprate (SC) was significantly higher than other PEs in comparison to control, and was selected for further investigation. All concentrations of SC (10-200 mM) demonstrated enhanced flux of ZMR in comparison to control. The highest flux (13 folds higher than control) was achieved for the formulation with highest SC concentration (200 mM). The relative BA of ZMR formulation containing SC (PO-SC) in plasma at a dose of 10 mg/kg following oral administration in rats was 317.65% in comparison to control formulation (PO-C). Besides, the AUC0-24 h of ZMR in the lungs following oral administration of PO-SC was 125.22 +/- 27.25 ng hr ml(-1) with a Cmax of 156.00 +/- 24.00 ng/ml reached at 0.50+/-0.00 h. But, there was no ZMR detected in the lungs following administration of control formulation (PO-C). The findings of this study indicated that the oral formulation PO-SC containing ZMR and SC was able to enhance the BA of ZMR in plasma to an appropriate amount that would make ZMR available in lungs at a concentration higher (>10 ng/ml) than the IC50 concentration of influenza virus (0.64-7.9 ng/ml) to exert its therapeutic effect.
Administration, Oral ; Animals ; Biological Availability* ; Caco-2 Cells ; Humans ; Influenza, Human ; Inhibitory Concentration 50 ; Lung* ; Orthomyxoviridae ; Permeability ; Plasma* ; Rats* ; Sodium ; Sodium Cholate ; Zanamivir*

We conducted two studies to determine the effect of gender, gonadectomy (GDX) on growth and plasma cholesterol levels in pigs. In experiment 1, five sham-operated and five GDX female Landrace pigs (26 kg) were allowed to have free access to water and feed up to market weight (approximately 100 kg). Body weight and feed consumption were recorded biweekly, and daily body weight gain, daily feed intake and feed efficiency (gain/feed) were calculated during the feeding period. In experiment 2, 10 male (26 kg) and 10 female (26 kg) Landrace pigs were used; five male and five female pigs were assigned to sham-operated or GDX. Pigs were allowed to have free access to water and a diet without added cholesterol (Table 1) until they were 6 months old (male 104 and female 98 kg) and thereafter they were fed a hypercholesterolemic diet (Table 1) containing 0.5% cholesterol and 0.1% cholate for 10 days. GDX of female pigs increased average daily gain (P<0.05), compared with their sham-operated counterparts during the growing-finishing period, but had no effect (P>0.05) on feed efficiency. Plasma cholesterol levels in pigs fed a hypercholesterolemic diet for 10 days were much higher (P<0.05) in females than in males (161 vs 104 mg/100 mL plasma), and were increased by GDX only in male pigs. HDL-cholesterol/LDL+VLDL-cholesterol ratio appeared to be higher in males than in females, and was not influenced by GDX in either sex. Results suggested that the lower growth rate of female pigs than their male counterparts is attributable to the ovarian activity, and the lower plasma cholesterol level in male than in female pigs fed a hypercholesterolemic diet is due to the testicular activity.
Body Weight ; Cholates ; Cholesterol ; Diet ; Female ; Gonadal Steroid Hormones ; Humans ; Male ; Plasma ; Swine ; Water

This study investigated that the antioxidative effect of freeze-dried cranberry powder against protein and lipid oxidation and ameliorative effect of serum lipid profile in rat fed atherogenic diet. Six weeks old male Sprague-Dawley rats were divided into the following four groups: normal diet group with 5% corn oil (control), atherogenic diet group with 5% corn oil, 10% lard, 1% cholesterol, and 0.5% sodium cholate (HFC), atherogenic plus 2% cranberry powder diet group (HFC + C2), and atherogenic plus 5% cranberry powder diet group (HFC + C5), and respective diet and water were fed daily for 6 weeks. After the experimental period, the serum lipid profile, such as total cholesterol, HDL-cholesterol, LDL-cholesterol, and triglyceride, ferric reducing ability of plasma (FRAP), plasma phenolics content, superoxide dismutase (SOD) activity, serum protein carbonyl and thiobarbituric acid reactive substances (TBARS) levels were examined. Total phenolic compound and total flavonoid levels in freeze-dried cranberry powder were 9.94 mg/g and 8.12 mg/g, respectively. Serum total cholesterol and LDL-cholesterol levels were not significantly different for cranberry powder treatment, but serum HDL-cholesterol level was significantly increased in HFC + C5 group compared with HFC group. Plasma FRAP value tended to be increased by cranberry powder treatment though there was no significant difference. Plasma total phenol concentrations and SOD activities were not significantly different among all groups. Serum protein carbonyl and TBARS levels were significantly decreased in HFC + C5 group compared with HFC group. Overall results suggested that freeze-dried cranberry powder might have the serum lipid improving effect, as well as antioxidative effect demonstrated by its protective effect against protein and lipid oxidation.
Animals ; Biomarkers ; Cholesterol ; Corn Oil ; Diet ; Diet, Atherogenic ; Dietary Fats ; Humans ; Male ; Oxidative Stress ; Phenol ; Plasma ; Rats ; Rats, Sprague-Dawley ; Sodium Cholate ; Superoxide Dismutase ; Thiobarbiturates ; Thiobarbituric Acid Reactive Substances ; Vaccinium macrocarpon ; Water

The effects of taurine on plasma and liver cholesterol, erythrocyte ouabain sensitive Na efflux and platelet aggregation were examined in Sprague Dawley rats fed control or 0.5% cholesterol with 0.2% cholate diet. Plasma and liver levels of total cholesterol were increased significantly (p<0.05) in rats fed cholesterol diet compared to the control, and taurine significantly decreased the elevated plasma level of cholesterol in rats fed cholesterol diet (p<0.05). HDL-cholesterol was decreased in groups fed the cholesterol diet regardless of taurine supplementation and the difference between groups with and without cholesterol was significant (p<0.01). Plasma triglyceride was decreased and liver triglyceride was increased both significantly (p<0.05) in rats fed cholesterol compared to the control. Plasma and liver triglyceride in rats fed taurine was decreased significantly compared to the control (p<0.05). Intracellular Na tended to be lower in rats fed cholesterol or taurine and higher in rats fed cholesterol plus taurine compared to the control. Na efflux through Na-K ATPase and the passive leak of Na was somewhat reduced in rats fed cholesterol or taurine and was augmented in rats fed cholesterol plus taurine compared to the control, which showed a similar trend to the intracellular Na. Taurine supplementation caused a suppression of Na efflux in groups fed control diet and restored the suppressed Na efflux in groups fed cholesterol. Platelet aggregation was significantly decreased in the group fed taurine compared to the control (p<0.05) and the group fed cholesterol plus taurine was also a little lower in aggregation than the group fed cholesterol. Microscopic examination showed that taurine prevented fatty liver in rats fed cholesterol diet. Taurine known for stimulating Na-K ATPase in some cell types rather decreased erythrocyte ouabain sensitive Na-K ATPase in the present study. Taurine had hypolipidemic and hypocholesterolemic effects and inhibited platelet aggregation which may be favorable for prevention of cardiovascular diseases.
Adenosine Triphosphatases ; Animals ; Blood Platelets* ; Cardiovascular Diseases ; Cholates ; Cholesterol ; Diet ; Erythrocytes* ; Fatty Liver ; Liver* ; Ouabain* ; Plasma* ; Platelet Aggregation* ; Rats ; Rats, Sprague-Dawley* ; Taurine* ; Triglycerides

OBJECTIVETo prepare tanshinone transfersome (TTs) and evaluate its deformability.

METHODSThe transfersomes were prepared by film dispersion method followed by sonication, and their physical properties, morphology, content, entrapment efficiency, particle size, polydispersity, and Zeta potential were investigated. The stability and deformability of TTs were studied. Liposomes with different molar ratios of cholate and lecithin were compared for their permeability under external pressure.

RESULTSThe prepared TF were spherical vesicles with content of 1.0192+/-0.075 mg/ml, entrapment efficiency of (62.3+/-0.08)%, particle size of 110 nm, polydispersity of 0.19 and Zate potential of -15.0 mV. The TTs remained stable during light-proof preservation for 3 months at 4 degrees C, and sodium cholate contributed to the flexibility of the lecithin vesicles.

CONCLUSIONTTs prepared by film dispersion method has good entrapment efficiency and stability. The vesicles possess high deformability in relation to the molar ratio of sodium cholate to lecithin and the external pressure.

Diterpenes, Abietane ; Drug Compounding ; methods ; Lecithins ; chemistry ; Liposomes ; chemistry ; Microscopy, Electron, Transmission ; Particle Size ; Phenanthrenes ; chemistry ; Sodium Cholate ; chemistry ; Technology, Pharmaceutical ; methods ; Temperature

AIMTo prepare capsaicin transfersomes and evaluate them in vitro and in vivo.

METHODSCapsaicin transfersomes were prepared by high shear dispersing machine and evaluated by entrapment efficiency, release rate, in vitro skin permeation and distribution in different tissues in vivo.

RESULTSCapsaicin transfersomes were composed of single unilamellar vesicles with an average diameter of 150.6 nm. Capsaicin entrapment efficiency increased distinctly with increasing of concentration of lecithin and entrapment efficiency is 96.7% while concentration of lecithin to 8%. Cumulative release amount of capsaicin is in direct proportion to the ethanol concentration in the receptor medium. In vitro capsaicin cumulative penetration amount showed higher levels in transfersomes than cream and suspension in rat abdominal skin. Abdominal skin cumulative penetration amount in vitro of capsaicin transfersomes in mouse was significantly higher than that from rat and men. In the same way, abdominal skin epidermal membrane cumulative penetration amount in vitro of capsaicin transfersomes was significantly higher than that from derma and full skin in human abdominal skin. The capsaicin tissue distribution of capsaicin injection by multiple celiac injections in rats is different: bone > plasma > skin > muscle. There is a similar result by multiple thigh topical application of capsaicin transfersomes: bone > skin > plasma > muscle.

CONCLUSIONEntrapment efficiency of capsaicin transfersomes reached the criterion of China Pharmacopoeia (> 80%) and capsaicin skin penetration can be increased by capsaicin transfersomes. It should be noted that the diverse characters and levels of skin may probably affect the permeating capability of capsaicin. Capsaicin tissue distribution in bone and muscle is similar and is different in plasma and skin by multiple injections and topical skin apply.

Administration, Cutaneous ; Animals ; Anti-Inflammatory Agents, Non-Steroidal ; administration & dosage ; pharmacokinetics ; Capsaicin ; administration & dosage ; pharmacokinetics ; Drug Carriers ; Drug Delivery Systems ; Humans ; Lecithins ; chemistry ; Male ; Mice ; Particle Size ; Rats ; Rats, Sprague-Dawley ; Skin Absorption ; Sodium Cholate ; chemistry ; Tissue Distribution

Animals ; Blood Glucose ; metabolism ; Colon ; metabolism ; ultrastructure ; Hypoglycemic Agents ; pharmacokinetics ; Immunohistochemistry ; Insulin ; pharmacokinetics ; Intestinal Absorption ; drug effects ; Male ; Microscopy, Electron ; Rats ; Rats, Sprague-Dawley ; Sodium Cholate ; pharmacology

AIMTo study the relationship between cellular membrane fluidity and relative bioavailability (Fr) of protein and peptide drugs combined with absorption enhancers after pulmonary administration in rats.

METHODSA series of model drug salmon calcitonin (sCT) solutions with 6 absorption enhancers (Brij78, sodium cholate, sodium caprylate, 2-hydroxypropyl-beta-cyclodextrin, lecithin and chitosan) were prepared and then delivered to rats by pulmonary route. Serum drug concentration was determined by radioimmunoassay method. Using the techniques of electron spin resonance and fluorescence polarography, the effects of enhancers on pulmonary cellular membrane fluidity were investigated.

RESULTSFr values of sCT solution with some absorption enhancers (Brij78, sodium cholate, sodium caprylate, lecithin and chitosan) were significantly higher than those without enhancers. Brij78, lecithin and sodium caprylate, not only increased membrane lipid fluidity but also loosed the constitution of membrane protein. The effect of sodium cholate on membrane protein was low. Lipid fluidity was reduced and protein constitution was changed markedly, after pulmonary cellular membrane was treated by 0.5% chitosan solution. This result showed that the absorption enhancing of chitosan mainly came from its effects on membrane protein. Corresponded with lower Fr after pulmonary administration, 2-hydroxypropyl-beta-cyclodextrin (0.5% and 3%) had not significant effects on both lipid fluidity and protein constitution.

CONCLUSIONThe effects of enhancers on pulmonary absorption of peptide drugs in vivo might be investigated on the grounds of determination of cellular membrane fluidity in vitro.

Absorption ; Animals ; Biological Availability ; Calcitonin ; administration & dosage ; pharmacokinetics ; Caprylates ; pharmacology ; Cell Membrane ; metabolism ; Chitin ; analogs & derivatives ; pharmacology ; Chitosan ; Drug Synergism ; Lung ; metabolism ; Male ; Membrane Fluidity ; drug effects ; Membrane Proteins ; drug effects ; Molecular Conformation ; Phosphatidylcholines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sodium Cholate ; pharmacology

PURPOSE: Cytotoxicity of the bile acids on colon cancer cell lines was studied to know which bile acid was most cytotoxic to colonic mucosal epithelium. We performed agarose gel electrophoresis whether this toxicity was caused by detergent effect of the bile acids or by apoptotic pathway. MATERIALS AND METHODS: HT29, LoVo, SW620 colon cancer cell lines were exposed to lithocholate, cholate, deoxycholate and chenodeoxycholate with 50, 100, 150, 200, 250, 300 pM as final concentration in DMEM culture media for short time (for 2 hours) and for long time (for 5 days). Agarose gel electrophoresis was performed on each colon cancer cell lines (HT29, LoVo, SW620, SW480) after 1, 2, 3, 4, 5 days exposure to deoxycholate with 150 pM concentration to detect intemucleosomal fragmentation. RESULTS: There was no toxicity after short time exposure in all bile acids concentration and in all colon cancer cell lines. Of the bile acids, deoxycholate was most toxic for all colon cancer cell lines. And DNA fragmentation was noticed after 2 days exposure with deoxycholate. Only LoVo cell line showed apoptotic DNA pattern after 4 days of exposure with deoxycholate. CONCLUSION: Bile acids (especially deoxycholate) are suggested to be possible agents to cause apoptosis in colonic mucosal epithelium.
Apoptosis ; Bile Acids and Salts* ; Bile* ; Cell Line* ; Chenodeoxycholic Acid ; Cholates ; Colon* ; Colonic Neoplasms* ; Culture Media ; Deoxycholic Acid ; Detergents ; DNA ; DNA Fragmentation ; Electrophoresis, Agar Gel ; Epithelium ; Lithocholic Acid

Ischemia causes tissue necrosis in a wide variety of pathologic conditions. Permanent deprivation of blood flow is lethal to any tissue and the prudent therapy for ischemia unquestionably is reperfusion. While reperfusion is necessary to reverse the progression towards ischemic death, reperfusion is also thought to be accompanied by its own component of injury. Oxygen free radicals, formed during ischemia/reperfusion, have been proposed as one of the main causes of reperfusion injury. Free radical attacks on biological membrane, such as mitochondria and endoplasmic reticulum, and can lead to the oxidative destruction of the polyunsaturated fatty acids of the membranes through lipid peroxidation. However, direct association between microsomal lipid peroxidation in vivo after ischemia/reperfusion and changes in secretory function and drug metabolism on the liver have not been established. Therefore, present study was performed to evaluate the hepatic secretory function and the hepatic microsomal drug metabolizing enzyme activity after ischemia/reperfusion preparation in rat liver. Further, the effect of oxygen free radical scavengers was investigated. The animals were divided into sham operation group and ischemia/reperfusion group. The ischemia/reperfusion group was subdivided into non-treated control and treated (with superoxide dismutase, allopurinol, alpha-tocopherol, deferoxamine) groups. Hepatic ischemia was produced by clamping the left branches of portal vein and hepatic artery, resulting in complete ischemia to the median and left lobes while the right lobes remained per Fused to prevent intestinal congestion. Reperfusion was permitted by declamping after 1 hour. After 1 or 5 hours of reperfusion, bile was collected, blood was obtained from abdominal aorta, and liver microsomes were isolated. The results are as follows. Serum aminotransferase was increased 15~20 times by ischemia/reperfusion. However, this increase was attenuated by free radical scavengers, especially 5 hours of reperfusion. The wet weight-to-dry weight ratio of the liver was significantly increased by ischemia/reperfusion. alpha-tocopherol pretreatment minimized the increase of ratio. Malondialdehyde level in the liver microsomal fraction was significantly increased after ischemia/reperfusion, but this increase was attenuated by scavenger pretreatment, especially alpha-tocopherol. Bile flow and cholate output but not the bilirubin output, were decreased after ischemia/reperfusion. The free radical scavenger pretreahnent restored the secretion significantly. Cytochrome P-450 content was significantly decreased after ischemia/reperfusion and ameliorated by free radical scavenger pretreatment. NADPH cytochrome P-450 reductase activity and aminopyrine N-demethylase activity were also decreased and improved by free radical scavengers pretreatment. These results indicate that ischemia/reperfusion deteriorates the hepatic secretory function as well as hepatic microsomal drug metabolizing enzyme activity, and the oxygen free radical scavengers attenuate the functional changes of the liver induced by ischemia/reperfusion.
Allopurinol ; alpha-Tocopherol ; Aminopyrine N-Demethylase ; Animals ; Aorta, Abdominal ; Bile ; Bilirubin ; Cholates ; Constriction ; Cytochrome P-450 Enzyme System ; Endoplasmic Reticulum ; Estrogens, Conjugated (USP) ; Fatty Acids, Unsaturated ; Free Radical Scavengers ; Free Radicals ; Hepatic Artery ; Ischemia ; Lipid Peroxidation ; Liver ; Malondialdehyde ; Membranes ; Metabolism* ; Microsomes, Liver ; Mitochondria ; NADPH-Ferrihemoprotein Reductase ; Necrosis ; Oxygen ; Portal Vein ; Rats* ; Reperfusion ; Reperfusion Injury ; Superoxide Dismutase