[Effect of emodin lipid nano-microbubble on MAPK signal pathway and inflammation cytokine in AT-II cells by mechanical stretch].

OBJECTIVE: To investigate the effect of emodin lipid nano-microbubble on MAPK signaling pathway and the level of inflammation cytokine in AT-II cells induced by mechanical stretch and its mechanism. METHODS: Emodin nanofibers, prepared by electrospinning with lecithin and PVP as carrier, were put into a seal bottle full of perfluoropropane, after they mixed with water and turned into self-assembled nano-microbubble. AT-II cells, separated and purified from primary rat AT-II cells, suffered 20% mechanical stretch for 4, 8, 16, 24, 48 h, and its protein expressions of p-P38/P38, p-ERK/ERK, p-JNK/JNK and the inflammation cytokine release levels of TNF-alpha, IL-1beta, IL-6 were detected by Western-Blot and ELISA. And further oberved the intervention effect of emodin lipid nano-microbubble on VILI. RESULTS: The protein expressions of p-P38, p-ERK, p-JNK were significantly increased in AT-II cells induced by mechanical stretch, and continuously evaluated from the 4-16 h, but the protein expressions of p38, ERK, JNK had no significant difference. The release levels of TNF-alpha, IL-1beta, IL-6 in AT-II cells had no change during 8 h, and they were gradually increased significantly in followed 16 h. In AT-II cells induced by mechanical stretch, due to intervention effect of emodin lipid nano-microbubble, the the protein expressions of p-P38, p-ERK, p-JNK and the inflammation cytokine release levels of TNF-alpha, IL-1beta, IL-6 were significantly decreased. CONCLUSION: Emodin lipid nano-microbubble shows protective effect on AT-II cells induced by mechanical stretch (VILI), and its mechanism may be related to down-regulation of protein expression of MAPK signaling pathway to regulate the release levels of inflammatory cytokine.

Electrospun drug-loaded core-sheath PVP/zein nanofibers for biphasic drug release.

Core-sheath nanofibers prepared using coaxial electrospinning were investigated for providing biphasic drug release profiles. With ketoprofen (KET) as the model drug, polyvinylpyrrolidone and zein as the sheath polymer and core matrix, respectively, the coaxial process could be carried out smoothly and continuously without any clogging of the spinneret. Scanning electron microscopy and transmission electron microscopy observations demonstrated that the nanofibers were linear with homogeneous structure and had a clear core-sheath structure with an average diameter of 730 ± 190 nm, in which the sheath had a thickness of ca. 90 nm. Differential scanning calorimetric and X-ray diffraction analyses verified that all the components in the core-sheath nanofibers were present in an amorphous state. Attenuated total reflectance Fourier transform infrared spectra demonstrated both the sheath and core matrix had good compatibility with KET owing to hydrogen bonding. In vitro dissolution tests showed that the nanofibers could provide an immediate release of 42.3% of the contained KET, followed by a sustained release over 10h of the remaining drug. The present study exhibited a simple and useful approach to systematically design and fabricate nanostructures using coaxial electrospinning for providing biphasic drug release profiles.

Receptor-mediated, tumor-targeted gene delivery using folate-terminated polyrotaxanes.

Safe and effective gene delivery is essential to the success of gene therapy. We synthesized and characterized a novel nonviral gene delivery system in which folate (FA) molecules were functioned as blockers on cationic polyrotaxanes (PR) composed of poly(ethylenimine) (PEI)(600)-grafted α-cyclodextrin rings linearized on polyethylene glycol to form FA-terminated PR-PEI(600) (FPP). The FA terminal caps of FPP target cell surfaces abundant in FA receptor (FR), a common feature of tumor cells. The structure of FPP was characterized by using (1)H nuclear magnetic resonance ((1)H NMR). The delivery particle was composed of chemically bonded PEG (4000), α-cyclodextrins (CD), and PEI (600 Da) at a molar ratio of 1:17:86.7, and the particle size and zeta potential of FPP/pDNA polyplexes were measured using dynamic light scattering. FPP/pDNA exhibited a lower cytotoxicity, strong specificity to FR, and high efficiency of delivering DNA to target cells in vitro and in vivo with the reporter genes. Furthermore, the FPP/DNA complex showed an enhanced antitumor effect in the nude mice compared with other delivery systems, such as PEI-25K. Together, these results suggest that FPP may be useful for gene therapy.

[Gene transfer system mediated by PEI-cholesterol lipopolymer with lipid microbubbles].

The properties of polyethyleneimine-cholesterol cationic lipopolymer (PEI-Chol) as gene carries and its gene transfer efficiency in vitro with lipid microbubbles were presented in this paper. PEI-Chol lipopolymer was synthesized by linking cholesterol chloroformate to the amino groups of branched poly(ethyleneimine) (PEI) of 1 800. The structure and molecular weight of PEI-Chol were confirmed by IR, 1H NMR and MADI-TOF-MS (matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry), respectively. The average molecular weight of PEI-Chol was approximately 2 000. The gene delivery system of bubble/PEI-Chol/DNA was constructed by mixed PEI-Chol/pDNA (N/P 10:1) complexes with lipid microbubbles (2-8 microm) which were prepared by DPPC, DSPE-PEG2000 and perfluoropropane with the reverse phase evaporation technique. pEGFP-Cl (enhanced green fluorescent protein) was used as report gene to investigate the DNA condensing ability of PEI-Chol lipopolymer by agarose gel electrophoresis. And their cytotoxicity and in vitro transfer efficiency of different complexes were compared with each other in A549 and MCF-7. The results indicated PEI-Chol lipopolymer can condense plasmid DNA when N/P ratio upto 4, PEI-Chol complexes and bubble/PEI-Chol/DNA complexes were nontoxic to A549 and MCF-7 when formulated at the N/P ratio of 10/1 as determined by MTT assay. This bubble/PEI-Chol/DNA delivery system provided good transfer efficiency with other desirable characteristics such as against-precipitation of plasma proteins. In conclusion, bubble/PEI-Chol/DNA complex is a novel non-viral gene delivery system.

[Preparation of self-emulsifying soft capsule and its pharmacokinetic in rats for epimedium flavonoids].

OBJECTIVE: To study the formulation optimization of epimedium flavonoids self-emulsifying drug delivery system and evaluate its effect in vitro and in vivo. METHODS: Based on the degree of emulsification and emulsifying time, the formulation optimization (i.e., screening of suitable oil phases, nonionic surfactants and co-surfactants) was made by the use of determination of the solubility, orthogonal design and construction of tertiary phase diagram. The dissolution of SEDDS was measured and its pharmacokinetic in rats was measured. RESULTS: The experimental results revealed the optimized formulation of the self-emulsifying drug delivery system which consisted of oleic acid as oil phase, Tween-80 as nonionic surfactant, and PEG400 as co-surfactant in the proportion of 2:4:4. The dissolution of SEDDS in water was more than 85% in 25 minutes, while that of the epimedium flavonoids capsule was less than 50% in 60 minutes. CONCLUSION: Application of the optimized formulation of epimedium flavonoids self-emulsifying drug delivery system could significantly increase the solubility of epimedium flavonoids in water and improve its bioavailability.

[Effects of Tankejing dry powder inhaler on the inflammatory damage in the lungs of mice infected by FM1].

OBJECTIVE: To study the effect of Tankejing Dry Powder inhaler on inflammatory in lung injury of mice infected by influenza virus (FM1). METHODS: Model mice infected by influenza virus FM1 were randomly divided into six groups: the nomal group, model group, low dose, medium dose and high dose of Tankejing Dry Powder inhaler group,and Ribavirin group. The following indices including the change of NF-kappaB in lung tissue at 24 h, the changes of TNF-alpha, IL-1beta, IL-6 in lung tissue, the lung index and the death rate were observed. RESULTS: Compared with model group, medium and high dosages of Tankejing Dry Powder inhaler could modulate the expression of NF-kappaB, reduce the level of TNF-alpha, IL-beta, IL-6 in lung tissue (P < 0.05), and reduce the lung index and the death number of animals within 14 days (P < 0.05). CONCLUSION: Tankejing Dry Powder inhaler may reduce the inflammatory injury caused by influenza virus FM1 through regulating the expression level of NF-kappaB and cytokine.

[Anti-viral effects of Tankejing preparations against influenza virus A].

OBJECTIVE: To investigate the anti-viral effects of Tankejing preparations (including solutions, dry power inhalation and powder) against influenza virus A in vitro and the relationship between its anti-viral effect and preparations. METHODS: The inhibitive effect of different drug-added ways of Tankejing extracts against human influenza virus A (H3N2) in vitro were assayed by crystallized purple staining method with ribavirin as positive reference drug. Then we compared the anti-viral actions of different kinds of Tankejing preparations. At last, we carried on serum pharmacodynamics to test and verify the effect. RESULTS: Tankejing extracts had an effect on comprehensive inhibition and an inhibition on virus proliferation after adsorption. When the concentration was 0.062 g/mL, the inhibition rate of solution dry power inhalation and powder were 43.50%, 41.50% and 37.36%, respectively. The anti-viral effect of dry powder inhalation was better than power's in serum pharmacokinetis experiment. CONCLUSION: Tankejing preparations display an action against human influenza virus A in vitro. Those preparation with better solubility have greater anti-viral effect.

[Effects of epimedium total flavonoids phytosomes on preventing and treating bone-loss of ovariectomized rats].

OBJECTIVE: To observe the effects of Epimedium total Flavonoids Phytosomes on preventing and treating bone-loss of the castrate osteoporosis rat model. METHOD: The osteoporosis model was established with 4-month-odl panther's rats, their ovaries on both sides castrated. Dual energy X-ray scanning was used to determine the bone density, and immunity and ELASA were used to assay concentration of estradiol and IL-6 in serum respectively, then determine their effect. RESULT: The BMP and E2 of high dosage group nilestriol group and normal group are higher than those of model group (P < 0.01), while their content of IL-6 is apparently lower than that of model group(P < 0.01). CONCLUSION: The osteoporosis model was established successfully and the using of EFP can improve the bone density, enhance E2 level and decrease the IL-6 concentration in serum.