[Effects of Serum Containing Qinbai Qinfei Concentrated Pellets on Expressions of NLRP3 Inflammasome in RAW264.7 Cells Infected with Mycoplasma pneumonia].

Objective: To investigate the effects of serum containing Qinbai Qingfei concentrated pellets on expressions of NLRP3 inflammasome in RAW264. 7 cells infected with Mycoplasma pneumoniae( MP) IL-1ß. Methods: RAW264. 7 cells were randomly divided into normal group, MP model group and serum containing Qinbai Qinfei concentrated pellets group. RAW264. 7 cells and MP strain were cultured utilizing normal methods, preparation of serum containing Qinbai,with 1∶ 10 multiplicity of infection( MOI) of MP stimulation on RAW264. 7 cells; cells of each group were collected at 8,16,24 h respectively. The expressions of NLRP3,ASC and Caspase-1 mRNA were detected by the method of FQ-PCR. The expressions of NLRP3,ASC and Caspase-1 p20 protein were detected by Westernblot. The content of IL-1ß in the supernatant was measured by ELISA. Results: Compared with the normal group, he levels of NLRP3,ASC and Caspase-1 mRNA were significantly increased in the model group at 8,16,24 h respectively( P < 0. 05 or P < 0. 01); while the levels of NLRP3,ASC and Caspase-1 p20 protein were increased significantly( P < 0. 05 or P < 0. 01) at 16,24 h, and the levels of IL-1ß were increased at significantly( P < 0. 01) 24 h. Compared with the model group, the levels of NLRP3,ASC and Caspase-1 mRNA were significantly reduced in serum containing Qinbai Qinfei concentrated pellets group at 16,24 h( P < 0. 05 or P < 0. 01); the expressions of NLRP3,ASC,Caspase-1 p20 protein and the content of IL-1ß were all decreased at 24 h.Conclusion: The mechanism of antiMycoplasma pneumoniae action of Qinbai may be related to the down-regulation of NLRP3 inflammasome expressions.

Preparation of a liposomal delivery system and its in vitro release of rapamycin.

The aim of this study was to prepare a liposomal delivery system for rapamycin and study its in vitro release characteristics. The results may provide a foundation for the further development of a liposomal delivery system for rapamycin and the establishment of a new active treatment method targeted towards the cellular components of atherosclerotic plaques. The ethanol injection method was used to prepare rapamycin-containing liposomes. The formulation was optimized by orthogonal design, and the degree of rapamycin release by the liposomes was measured by the reverse dialysis method. Orthogonal testing showed that the optimum formulation had a phospholipid concentration of 4%, a phospholipid-cholesterol mass ratio of 8:1, a drug-lipid mass ratio of 1:20 and an aqueous phase pH of 7.4. Rapamycin-containing liposomes with an encapsulation efficiency of 82.11±2.13% were prepared, and the in vitro release of rapamycin from the liposomes complied with a first-order kinetic equation. In conclusion, the formulation was optimized, the prepared liposomes had a high rapamycin encapsulation rate and good reproducibility, and their in vitro release had a certain delayed-release effect.

[Liposomal encapsulation of free anthraquinones in rhizoma et radix rhei and its quality control].

OBJECTIVE: To encapsulate the free anthraquiones in Rhizoma et Radix Rhei (rhein, chrysophanol, physcione, emodin and aloeemodin) in liposomes and characterize the liposomes. METHODS: The liposomes were separated from free drug with sephadex G-50 with HPLC determination of the free anthraquinones in liposomes, then its entrapment efficiency was calculated. Ethanol injection method was used to prepare the liposomes with the addition of a suitable amount Ca2+ in the medium. The influence of Ca2+ on the entrapment efficiency of free anthraquinones in Rhizoma et Radix Rhei in liposomes has been studied. Dynamic laser scatterometer and transmission electron microscopy were used to study the sizes and morphology of the liposomes. RESULTS: The entrapment efficiency of rhein, chrysophanol, physcione, emodin and aloeemodin in liposomes were 27.63%, 85.16%,100.14%, 99.77% and 66.16%, respectively. The total entrapment efficiency of free anthraquinones in Rhizoma et Radix Rhei was 80.67%. Ca2+ could greatly promote the liposomal encapsulation of free anthraquinones in Rhizoma et Radix Rhei from 13.98% - 23.78% to 81.42% - 89.43%. The liposomes were white spheres and its average size was mainly concentrated on 340 nm. CONCLUSION: The entrapment efficiency of the liposomal encapsulation of free anthraquinones in Rhizoma et Radix Rhei is high. The quality control method is simple and rapid with good repeatability.

Degradation kinetics of water-insoluble lauroyl-indapamide in aqueous solutions: prediction of the stabilities of the drug in liposomes.

The aim of this study was to explore the degradation kinetics of water-insoluble lauroyl-indapamide in solutions and predict the stabilities of lauroyl-indapamide encapsulated in liposomes. Buffer-acetone (9:1) was used as the reaction solution and the reaction temperature was maintained at 60 degrees C. The correlation of the apparent degradation constants (k(obs)) of lauroyl-indapamide in liposomes and in buffer-acetone solutions at different pH has been explored. The degradation of lauroyl-indapamide in solutions was found to follow pseudo-first-order kinetics and was significantly dependent on the pH values. Lauroyl-indapamide was the most stable at pH 6.8, increasing or decreasing the pH of the solutions would decrease its stabilities. Buffer concentration had some effects on the stabilities of lauroyl-indapamide. The degradation active energies Ea were 68.19 kJ x mol(-1), 131.75 kJ x mol(-1) and 107.72 kJ x mol(-1) at pH3.6, 6.8 and 12 respectively in acetone-free buffer solutions (0.05M) calculated according to the Arrhenius equation with the extrapolation method. The apparent degradation constants (kobs) of lauroyl-indapamide in liposome and in buffer-acetone (9:1) solutions showed a good correlation at different pH levels, which indicates that the stabilities of the drug that dissolved in acetone-buffer mixture solutions can be used to predict the stabilities of the drug in liposomes as well.

HPLC determination of andrographolide in rat whole blood: study on the pharmacokinetics of andrographolide incorporated in liposomes and tablets.

A sensitive and simple high-performance liquid chromatographic method with UV detection was developed and validated for the determination of andrographolide in rat whole blood. Carbamazepine was employed as internal standard and the blood sample was extracted with chloroform. Chromatographic separations were achieved on a Chromasil ODS column (250 x 4.6 mm, 5 microm). The mobile phase was consisted of methanol-water (52:48, v/v) and delivered at 0.8 mL/min. The detection wavelength was set at 225 nm. The calibration curve had a good linearity in the range 0.053-530 microg/mL in rat whole blood with its correlation coefficient being 0.996. The extraction recovery of andrographolide was ranged from 65.7 to 72.6%. The intra-day and inter-days repeatabilities were below 4.2% in terms of the percentage of relative standard deviation (RSD). The method was used to provide data on the pharmacokinetics of the drug in rats. The data obtained was processed using the 3P87 pharmacokinetic program. The results showed that the disposition of andrographolide after intravenous administration of liposomal andrographolide conformed to a two-compartment open model with alpha = 4.75 x 10(-2) +/- 2.41 x 10(-3) min(-1), beta = 3.16 x 10(-3) +/- 1.58 x 10(-4) min(-1), V(c) = 174.67 +/- 13.97 mL, k(21) = 1.60 x 10(-2) +/- 8.12 x 10(-4) min(-1), k(10) = 9.38 x 10(-3) +/- 5.62 x 10(-4) min(-1), k(12) = 2.53 x 10(-2) +/- 1.27 x 10(-3) min(-1) and AUC(0-infinity) = 1525.47 +/- 92.35 microg min/mL. For the intragastric administration of andrographolide tablets, the disposition of andrographolide followed a one-compartment open model with k(e) = 6.78 x 10(-3) +/- 3.53 x 10(-4) min(-1), k(a) = 3.69 x 10(-2) +/- 4.68 x 10(-3) min(-1), T(max) = 59.69 +/- 3.61 min, C(max) = 1.62 +/- 0.11 microg/mL, V(c) = 1056.90 +/- 83.42 mL, AUC(0-infinity) = 348.75 +/- 24.41 microg min/mL.

Liposomal encapsulation of lauroyl-indapamide in the presence of divalent or trivalent cations.

Divalent or trivalent cations such as Ca2+, Ba2+, Mg2+, Zn2+, Fe2+, Al3+ and Fe3+ can cause a significant increase in the entrapment efficiency of lauroyl-indapamide in liposomes, from about 5% to more than 90%, which suggests that the presence of these ions plays an important role in the encapsulation of lauroyl-indapamide.

In vitro and in vivo studies of different liposomes containing topotecan.

Liposome as a carrier of topotecan (TPT), a promising anticancer drug, has been reported in attempt to improve the stability and antitumor activity of TPT. However, the biodistribution pattern of TPT liposome in vivo and PEG-modified liposome containing TPT have not been studied systemically. In this paper, the in vitro stability and in vivo biodistribution behavior of several liposomes containing TPT with different lipid compositions and PEG-modification were studied. Compared with the 'fluid' liposome (S-Lip) composed of soybean phosphatidylcholine (SPC), the 'solid' liposome (H-Lip) composed of hydrogenated soybean phosphatidylcholine HSPC decreased the leaking efficiency of TPT from liposome and enhanced the stability of liposome in fetal bovine serum (FBS) or human blood plasma (HBP). The results of biodistribution studies in S180 tumor-bearing mice showed that liposomal encapsulation increased the concentrations of total TPT and the ratio of lactone form in plasma. Compared with free TPT, S-Lip and H-Lip resulted in 5- and 19-fold increase in the area under the curve (AUC(0-->infinity)), respectively. PEG-modified H-Lip (H-PEG) showed 3.7-fold increase in AUC(0-->infinity) compared with H-Lip, but there was no significant increase in t(1/2) and AUC(0-->infinity) for PEG-modified S-Lip (S-PEG) compared with S-Lip. Moreover, the liposomal encapsulation changed the biodistribution behavior, and H-Lip and H-PEG dramatically increased the accumulation of TPT in tumor, and the relative tumor uptake ratios were 3.4 and 4.3 compared with free drug, respectively. There was also a marked increase in the distribution of TPT in lung when the drug was encapsulated into H-Lip and H-PEG. Moreover, H-PEG decreased the accumulation of TPT in bone marrow compared with unmodified H-Lip. All these results indicated that the membrane fluidity of liposome has an important effect on in vitro stability and in vivo biodistribution pattern of liposomes containing TPT, and PEG-modified 'solid' liposome may be an efficient carrier of TPT.