| Objective: To investigate the effects of particles with different size and Zeta potential characteristic in coptis-decoction on the berberine absorption in intestine. Ethyl cellulose particles with different size were prepared to simulation particles in copitis-decotions. The ethyl cellulose particles affect berberine absorption in intestine were evaluated by the in situ intestinal perfusion in rats, and the corresponding mechanism was discussed.Methods :(1) Preparation of copitis-decotions, using high-speed centrifugation gradient to remove the particles in larger size, then separate particles in the supernatant with weak-base anion-exchange resin in further by Zeta potential characteristic. The size and Zeta potential characteristics of particles in copitis-decotions were investigated.The Non-Particle-Coptis-Decoction was prepared by filter copitis-decotions with 0.45?m, 0.22 ?m, 0.1 ?m millipore filters according to priority. The constituent of particles in copitis-decotions were detected by phenol-sulfuric acid method and coomassie brilliant blue method. The effects of size and potential characteristics of particle on berberine absorption were evaluated by the in situ intestinal perfusion in rats.(2)Ethyl cellulose particles of different size were prepared using high voltage electrostatic spray. The size and morphlogy characteristics of ethyl cellulose particles were investigated by laser granularity analyzer and scanning electron microscope(SEM). The content of ethyl cellulose particles were detected by fluorescence spectrophotometry.Using the in situ intestinal perfusion in rats evaluation of different size of ethyl cellulose particles affect the absorption of berberine. The small intestine tissue cross-sectional section were made by freezing microtome and investigated by Hematoxylin-Eosin(HE)staning. The distribution of fluorescent tags nanoparticles in the small intestine tissue were observed by fluorescence microscope. The absorption characteristics of berberine on nanoparticles were investigated by the ultra high-speed centrifugation. The effect of ethyl cellulose on berberine absorption into the bloodstream evaluation were inverstigated by intestinal-vascular perfusion model, and the content of berberine in the blood was detected by high performance liquid chromatography-mass spectrometry(LC-MS).Results:(1) Particles P1, P2 and P3, with different size of(272 ±25.2) nm?(264.8±21.4)nm?(427.4±41.8) nm, and different surface potential of(-6.85±0.16) m V,(-18.20±0.71)m V and(-7.25±0.37) m V, were obtained from copitis-decotions by high-speed centrifugal separation and ion exchange chromatography. P1 and P2 with similar particle size distribution and differences potential value(P<0.05), while P1 and P3 with different size distribution(P<0.05) and similar potential value. There are not detected particles in Non-Particle-Coptis-Decoction. Although each segment of small intestine exhibited obvious berberine absorption, the jejunum exhibited the highest absorption rate constant(Ka) of(3.5879±0.0052)×10-4 /s and effective permeability coefficient(Papp)of(4.5294±0.0097)×10-5 cm/s. Preparation of Coptis-Decoction Perfusate,Non-Particle-Coptis-Decoction Perfusate and P1/P2/P3 Particle contained Berberine-K-R Perfusate, with Ka of(11.2344±1.0574) ×10-4 /s,(3.3584±0.0251) ×10-4/s,(11.8461±1.5677)×10-4 /s,(3.3325±0.0633)×10-4 /s and(5.8536±0.9701)×10-4 /s, and with Papp of(17.8949±3.0334)×10-5 cm/s,(6.5783±0.0576)×10-5 cm/s,(22.6375±2.0684)×10-5 cm/s,(4.5949±0.8019)×10-5 cm/s and(8.0824±1.0042)×10-5cm/s, respectively. It is suggest that solid particles have the ability to significantly promote the intestinal absorption of berberine. Particles P1 and P3 can significantly improve the intestinal absorption of berberine(P<0.05), especially the similar Ka and Papp of particle P1 with Coptis-Decoction.(2) Three kinds of nanoparticles P1, P2 and P3 are prepared by high-voltage electrostatic spray method, with size of(30.49±2.07) nm,(219.00±24.88) nm and(437.40±11.63) nm. Preparation of P1, P2 and P3 Particle contained Berberine-K-R Perfusate, with Ka of(1.6056±0.70270)×10-3 /s,(4.4379±0.5909)×10-3 /s and(1.6225±0.4719)×10-3 /s, and with Papp of (3.7723±0.9965)×10-4 cm/s,(8.2456±1.3453)×10-4 cm/s and(3.8572±0.9300)×10-4 cm/s,respectively. Particles P2 can significantly improve the intestinal absorption of berberine(P<0.05). Ethyl cellulose nanoparticles distributed in small intestinal were observed by Fluorescence microscopy. Nanoparticles have focused on goblet cells and lamina propria of intestine villi, also a small amount of particles in deep fossae. Every100 mg ethyl cellulose nanoparticles can adsorption berberine(122.48±21.06) ?g, and the adsorption rate was(10.37±1.8) %. Through the intestinal-vascular perfusion model,absorption into the bloodstream of berberine apparent permeability coefficient(Pblood) is(12.4373±1.9095)×10-6 cm/s.Conclusion: Particles in coptis-decotion can improve the berberine absorption in intestine signifivcantly. The size of distribution and surface potential of the particles may be responsible for the improvement of berberine absorption. The similar results are obtained by simulating size characteristic of particles in coptis-decotion. Berberine can be adsorbed on surface of ethyl cellulose nanoparticles, which may be absorbed into the blood jointly,to improve the bioavailability of berberine. |
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