To investigate the effect of methyleugenol on expression of MUC5AC in nasal mucosa of rats with allergic rhinitis (AR).Seventy-two Wistar rats were randomly divided into 6 groups:normal control group, AR group, loratadine group, low-dose methyleugenol group, middle-dose methyleugenol group and high-dose methyleugenol group with 12 rats in each group. AR was induced by intraperitoneal injection of ovalbumin in latter 5 groups. 10 mg loratadine q.d was given to rats in loratadine group by gavage; and 10 mg/kg, 20 mg/kg and 40 mg/kg methyleugenol were given by gavege q.d to rats in low-, middle-and high-dose methyleugenol groups, respectively. Nasal mucosa samples were obtained from rats at 1, 2, 4 and 6 weeks after drug intervention. The expression of MUC5AC protein and mRNA in nasal mucosa was detected by immunohistochemistry and real-time fluorescence quota PCR (RT-PCR), respectively.Compared with AR, the percentage of cells staining positively for MUC5AC protein and the relative quantity of MUC5AC mRNA in middle-and high-dose methyleugenol groups were significantly decreased after 2 and 4 weeks of drug intervention (<0.05), but no such decrease was observed in low-dose methyleugenol group at all time points (>0.05). The percentage of cells with positive expression of MUC5AC protein and mRNA in loratadine group were significantly decreased after 1 week of administration (<0.05). The percentage of cells with positive MUC5AC protein in middle-dose methyleugenol group was higher than that in loratadine group (<0.05) after 6 week of drug intervention, but the difference was not seen in high-dose group (>0.05). There was no significant difference in relative quantities of MUC5AC mRNA after 4 weeks of administration between high-and middle-dose methyeugenol groups and loratadine group (>0.05).Methyleugenol can attenuate AR through inhibiting the expression of MUC5AC mRNA and protein in nasal mucosa of AR rats.
Animals ; Dose-Response Relationship, Drug ; Down-Regulation ; drug effects ; Eugenol ; analogs & derivatives ; pharmacology ; Loratadine ; Mucin 5AC ; drug effects ; physiology ; Nasal Mucosa ; chemistry ; Ovalbumin ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Rhinitis, Allergic ; chemically induced ; drug therapy ; physiopathology

PURPOSE: The nasal mucosa is the first site to encounter pathogens, and it forms continuous barriers to various stimuli. This barrier function is very important in the innate defense mechanism. Additionally, inflammation of the nasal sinus is known to be a hypoxic condition. Here, we studied the effect of hypoxia on barrier function in normal human nasal epithelial (NHNE) cells. MATERIALS AND METHODS: The expression levels of various junction complex proteins were assessed in hypoxia-stimulated NHNE cells and human nasal mucosal tissues. We performed real-time polymerase chain reaction analysis, western blotting, and immunofluorescence assays to examine differences in the mRNA and protein expression of ZO-1, a tight junction protein, and E-cadherin in NHNE cells. Moreover, we evaluated the trans-epithelial resistance (TER) of NHNE cells under hypoxic conditions to check for changes in permeability. The expression of ZO-1 and E-cadherin was measured in human nasal mucosa samples by western blotting. RESULTS: Hypoxia time-dependently decreased the expression of ZO-1 and E-cadherin at the gene and protein levels. In addition, hypoxia decreased the TER of NHNE cells, which indicates increased permeability. Human nasal mucosa samples, which are supposed to be hypoxic, showed significantly decreased levels of ZO-1 and E-cadherin expression compared with control. CONCLUSION: Our results demonstrate that hypoxia altered the expression of junction complex molecules and increased epithelial permeability in human nasal epithelia. This suggests that hypoxia causes barrier dysfunction. Furthermore, it may be associated with innate immune dysfunction after encountering pathogens.
Anoxia/etiology/*metabolism ; Blotting, Western ; Cadherins/*analysis/genetics ; Epithelium/chemistry/pathology ; Humans ; Membrane Proteins/*analysis ; Nasal Mucosa/*chemistry/pathology/*secretion ; Permeability/*radiation effects ; RNA, Messenger/genetics/metabolism ; Real-Time Polymerase Chain Reaction ; Tight Junctions/*metabolism ; Zonula Occludens-1 Protein

In order to test the equilibrium solubility of puerarin in different solvents and solubilizer,cilia toxicity and irritation of these excipient, the balance method, toad in the ciliary body toxicity and rat nasal mucosa irritation were used respectively. Results showed that puerarin solubility was 56.44 g x L(-1) in combined solvent of 30% PEG200 and 10% Kolliphor HS 15. With normal saline solution as negative control and sodium deoxycholate as positive control, the effects of 30% PEG200, 30% PEG 400, 10% Kolliphor HS 15 and combination of 30% of PEG200 and 10% Kolliphor HS 15 on toad palate cilium were observed and cilia movement duration was recorded. The results indicated that there was no significant difference in cilia movement duration among 30% PEG200, 10% Kolliphor HS 15 and normal saline group. The rats long-term nasal mucous membrane irritation of 30% PEG 400, 10% Kolliphor HS 15, which had no cilia toxicity, was studied, with normal saline solution as negative control. There were no significant difference revealed on rat nasal mucosa epithelial thickness among 30% PEG 400, 10% Kolliphor HS 15 and normal saline. Above researches showed 30% PEG 400, 10% Kolliphor HS 15 was ideal for solubility of puerarin nasal drops and showed a lower cilia toxicity and irritation, and can be used as the solvent and solubilizer of puerarin nasal drops.
Administration, Intranasal ; methods ; Animals ; Anura ; Cilia ; chemistry ; Female ; Isoflavones ; chemistry ; Male ; Nasal Mucosa ; Polyethylene Glycols ; chemistry ; Rats ; Rats, Sprague-Dawley ; Solubility ; Solvents ; chemistry

The aim of this study is to prepare self-microemulsifying drug delivery system (SMEDDS) of the mixture of paeonol (Pae) and borneol (Bor). Solubility test, ternary phase diagrams and simplex lattice method were employed to screen and optimize the formulation of the mixture of Pae and Bor-loaded SMEDDS. After formed into microemulsions, the particle diameter (PD) was determined and a TEM was employed to observe the microemulsions' morphology. The contents of Pae and Bor were determined by gas chromatography. As a result, while ethyl oleate (EO) as the oil phase, cremophor EL35 (EL35) as surfactant and Transcutol HP (HP) as cosurfactant, the range of the microemulsion on the ternary phase diagram was larger than other combinations. And at a ratio of 20:45:35, the microemulsions' PD was about 34 nm and the polydispersity index (PI) was about 0.2. There were 16% of Pae, 2% of Bor, 16% of EO, 37% of EL35 and 29% of HP in the prepared SMEDDS. The preparation process of the Pae and Bor-loaded SMEDDS based on Xingbi Fang is simple and feasible. This study provides a reference for the researches on the related traditional Chinese medicine and the related components.
Acetophenones ; administration & dosage ; toxicity ; Administration, Intranasal ; Animals ; Bornanes ; administration & dosage ; toxicity ; Bufonidae ; Cilia ; drug effects ; Drug Combinations ; Drug Delivery Systems ; methods ; Drugs, Chinese Herbal ; administration & dosage ; toxicity ; Emulsions ; Ethylene Glycols ; chemistry ; Female ; Male ; Nasal Mucosa ; drug effects ; Oleic Acids ; chemistry ; Particle Size ; Polyethylene Glycols ; chemistry ; Solubility ; Surface-Active Agents ; chemistry

To investigate the effects of particle size, mPEG molecular weight, coating density and zeta potential of monomethoxyl poly(ethylene glycol)-poly(lactic-co-glycolic acid) (mPEG-PLGA) nanoparticles on their transportation across the rat nasal mucosa, mPEG-PLGA-NPs with different mPEG molecular weights (M(r) 1 000, 2 000) and coating density (0, 5%, 10%, 15%) and chitosan coated PLGA-NP, which loaded coumarin-6 as fluorescent marker, were prepared with the nanoprecipitation method and emulsion-solvent evaporation method, and determine their particle size, zeta potential, the efficiency of fluorescent labeling, in vitro leakage rate and the stability with the lysozyme were determined. The effects of physical and chemical properties on the transmucosal transport of the fluorescent nanoparticles were investigated by confocal laser scanning microscopy (CLSM). The result showed that the size of nanoparticles prepared with nanoprecipitation method varied between 120 and 200 nm; the size of nanoparticles prepared with emulsion-solvent evaporation method varied between 420 and 450 nm. Nanoparticles dispersed uniformly; the zeta potential of PLGA-NPs was negative; mPEG-PLGA-NPs was close to neutral; chitosan coated PLGA-NPs was positive; and the efficiency of fluorescent labeling were higher than 80%. In vitro leak was less than 5% within 4 h and nanoparticles were basically stable with lysozyme. The CLSM results show that the transportation efficiency of mPEG-PLGA-NPs with a high PEG coating density and high mPEG molecular weight was significantly higher than that of uncoated PLGA nanoparticles and also that of chitosan coated PLGA-NPs (P < 0.05). The hydrophilcity, zeta potential and particle size of nanoparticles play important roles on the efficiency of mPEG-PLGA nanoparticles to transport across the rat nasal mucosa.
Animals ; Biological Transport ; Chitosan ; chemistry ; Drug Carriers ; chemistry ; Female ; Male ; Microscopy, Confocal ; Molecular Weight ; Nanoparticles ; Nasal Mucosa ; metabolism ; Particle Size ; Polyesters ; chemistry ; pharmacokinetics ; Polyethylene Glycols ; chemistry ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the influence of natural borneol and synthetic borneol on mucosal permeability of Gardenia extract.

METHODTaken frog skin as a vitro model to study the vitro mucosal permeation the impacts of the natural borneols and synthetic borneols on the P(app) of the Jasminoidin were studied, and the effect of different borneols on the stability of Jasminoidin were investigated. Compared the 10 h accumulated infiltration rate of each group the effects of influence factors,such as C(Ge), C(B) and rotation speed on P(app) were investigated by using response surface method.

RESULTThe P(app) of Jasminoidin of natural borneol and synthetic borneol group were 1.44 fold and 1.77 fold of control group (P < 0.01). For two borneol groups, the results also showed a significant difference too (P < 0.05). Jasminoidin began to degrade about 8 h after the effect of frog skin for control group and synthetic borneol group, but was stable within 12 h for natural borneol group. The accumulated permeation rate of 10 h was same for different borneol groups. It was about 1.3 fold of control group. The C(Ge) had a salinence influence on the P(app) (P < 0.01) and C(B) had a salience influence on time-lag (P < 0.01).

CONCLUSIONBoth the natural borneol and synthetic borneol can accelerate the permeation of Jasminoidin and the synthetic borneol has stronger effect on the P(app). Both two different borneol can reduce the degradation effect of frog skin to Jasminoidin, but the natural borneol has a better protect effect on it. By using more natural borneol, the mucosal permeability of Gardenia extract can be increased, the time-lag can be reduced, and Jasminoidin has better stability.

Administration, Cutaneous ; Bornanes ; chemical synthesis ; pharmacokinetics ; Dosage Forms ; Drugs, Chinese Herbal ; chemistry ; Gardenia ; chemistry ; Iridoids ; pharmacology ; Mucous Membrane ; metabolism ; Nasal Mucosa ; metabolism ; Permeability ; Skin ; metabolism ; Skin Absorption

BACKGROUNDFascin, an actin binding protein, usually expressed at a low level in normal epithelium, but is significantly increased in transformed epithelial cells and several common carcinomas. In this study, we examined the expression of fascin by immunohistochemistry in sinonasal epithelium with chronic inflammation (control group), exophytic papilloma (EP), inverted papilloma (IP) with dysplasia and cancerated IP (including carcinoma in situ and invasive squamous cell carcinoma, SCC), and furthermore investigated the relationship between fascin expression and formation of malignant IP.

METHODSFascin expression was immunohistochemically detected using monoclonal antibody against fascin in 86 paraffin embedded tissues, including 10 cases of sinonasal mucosa with chronic inflammation, 10 of EP, 45 of IP with dysplasia (45 cases were divided into three groups: IP with mild dysplasia, IP with moderate dysplasia, and IP with severe dysplasia, 15 cases each), and 21 of cancerated IP.

RESULTSThe level of fascin expression was significantly higher in the neoplastic tissue than that in control group. Fascin expression increased gradually with the progression from sinonasal epithelium with chronic inflammation, IP with mild dysplasia, IP with moderate dysplasia, IP with severe dysplasia, to cancerated IP, and significant difference of fascin expression was observed between any two groups of the five.

CONCLUSIONPrecancerous lesions of IP exhibit elevated levels of fascin that may be associated with carcinogenesis of IP. Fascin may play a role in the formation of IP and EP.

Adult ; Aged ; Carrier Proteins ; analysis ; Cell Transformation, Neoplastic ; pathology ; Female ; Humans ; Immunohistochemistry ; Male ; Microfilament Proteins ; analysis ; Middle Aged ; Nasal Mucosa ; chemistry ; Nose Neoplasms ; chemistry ; pathology ; Papilloma, Inverted ; chemistry ; pathology ; Precancerous Conditions ; chemistry

AIMThe feasibility of intranasal brain targeting drug delivery system via the olfactory pathway from nose to brain was explored.

METHODSUsing gellan gum, a cation-sensitive gel forming excipient, huperzine A (Hup A) nasal in situ gel was prepared by pH gradient precipitation method. The pharmacokinetics of Hup A in blood and cerebrospinal fluid (CSF) after intranasal, intravenous and intragastric adminstration to rats was studied using cisternal cannulation for serial CSF sampling and femoral artery cannulation for serial blood sampling. The distributions of Hup A into rat brain tissues following intranasal dosing were compared with those after intravenous and intragastric dosing by tissue homogeneization. The therapeutics effects of Hup A nasal in situ gel on cognitive function were tested in mice and rats with Morris water maze, step down test and step through test.

RESULTSThe AUC(0-->6 h) value in plasma obtained after nasal administration was 0.94 of that after intravenous administration, but the AUC(0-->6 h) of CSF after nasal administration was 1.3 and 2.3 times of that after intravenous and intragastric administration. The AUC(0-->6 h), of cerebrum, hippocampus, cerebellum, left olfactory bulb and right olfactory bulb after nasal administration were 1.5, 1.3, 1.0, 1.2 and 1.0 of that after intravenous administration, 2.7, 2.2, 1.9, 3.1 and 2.6 times of that after intragastric administration, respectively. Intranasal adminintration of 17.5-35 microg x kg(-1) showed equal effects after oral adminintration of 70 microg x kg(-1) commercial tablets, which was in good agreement with the results of pharmacokinetics.

CONCLUSIONIntranasal administration of huperzine A nasal in situ gel significantly increased the distributions of Hup A into rat brain tissues, especially into cerebrum and hippocampus which should be the target areas of Hup A, and enhanced the brain targeting of Hup A.

Administration, Intranasal ; Alkaloids ; Animals ; Area Under Curve ; Biological Transport ; Brain ; metabolism ; Cattle ; Cognition ; drug effects ; Drug Compounding ; methods ; Guinea Pigs ; Injections, Intravenous ; Male ; Maze Learning ; drug effects ; Mice ; Models, Biological ; Nasal Mucosa ; metabolism ; Neuroprotective Agents ; administration & dosage ; pharmacokinetics ; pharmacology ; Particle Size ; Polysaccharides, Bacterial ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Sesquiterpenes ; blood ; cerebrospinal fluid ; pharmacokinetics

OBJECTIVETo study Xionggui nasal sprays and its evaluation in release in vitro and absorption in vivo.

METHODEstablishing the best prescription of Xionggui nasal sprays through orthogonal design methods, The in vitro release action of Xionggui nasal sprays was studied using dynamic dialyse method. The in vivo rat nasal recirculation methods were used to study the rule of Xionggui nasal sprays absorption.

RESULTThe optimum prescription was: Pemulen TR-1 0.35%, EDTA 0.2%, PEG400 1%, xanthan gum 0.2%; trolamine: right amount(adjust pH). Its release in vitro and absorption in vivo meet to Higuchi distribution.

CONCLUSIONThe preparation method of Xionggui nasal sprays was appropriate. The release of drug and its uptake was well correlated.

Absorption ; Administration, Intranasal ; Aerosols ; Angelica sinensis ; chemistry ; Animals ; Drug Carriers ; Drug Combinations ; Drug Compounding ; methods ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacokinetics ; Edetic Acid ; Emulsions ; Ethanolamines ; Female ; Ligusticum ; chemistry ; Male ; Nasal Mucosa ; metabolism ; Plants, Medicinal ; chemistry ; Polyethylene Glycols ; Polysaccharides, Bacterial ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the protective effect of purariae isoflavone on apoptosis cells of atrophic nasal mucosas in ovariectomized rats.

METHOD60 rats were divided into four groups as control, ovariectomized, ovariectomized + nylestriol (O + N) and ovariectomized + purariae isoflavone (O + P), each with 15 rats. Earlier apotosis cells of mucosas taken from nasal septum were measured with flow cytometry.

RESULTCompared with control group, and the number of apoptosis cells of mucosas increased after being ovariectomized,and the number of apoptosis cells of mucosas in O + N and O + D group didn't change.

CONCLUSIONNylestriol and purariae isoflavone might have effects on protecting cells of mucosas from lacking of estrogen by decreasing apoptosis cells in ovariectomized rats.

Animals ; Apoptosis ; drug effects ; Epithelial Cells ; pathology ; Estradiol Congeners ; pharmacology ; Female ; Isoflavones ; isolation & purification ; pharmacology ; Nasal Mucosa ; pathology ; Ovariectomy ; Plants, Medicinal ; chemistry ; Protective Agents ; isolation & purification ; pharmacology ; Pueraria ; chemistry ; Quinestrol ; analogs & derivatives ; pharmacology ; Rats ; Rats, Sprague-Dawley