ABSTRACT
OBJECTIVE：To study the mechanism of Periplaneta americana extract degreasing cream and CⅡ-3（shorted for “degreasing cream ”and“CⅡ-3”）reversing the multi-drug resistance of human HepG 2/ADM cells. METHODS ：MTT assay was used to investigate the toxicity effects of different concentrations of sorafenib （positive control ），degreasing cream and C Ⅱ-3 on HepG2/ADM cells ，then IC 20 was calculated. The experiment was divided into sensitivity drug ，drug-resistance group ，sorafenib group，degreasing cream group and C Ⅱ-3 group. HepG 2 cells were included in sensitivity group ，and HepG 2/ADM cells were included in the latter 4 groups. Sensitivity group and drug-resistance group were treated with routine medium ，and other 3 groups were treated with relevant medicine （IC20 as drug concentration ）. The content of ADM in HepG 2/ADM cells was determined by Laser scanning confocal microscopy. The expression of apoptosis-related protein as Bcl- 2 and Cleaved-Caspase- 9 p37 were detected by Western blotting assay. RT-qPCR and immunocytochemistry were adopted to detect mRNA and protein expressions that related to multidrug resistance [P-gp （expression produce of MDR1 gene），LRP，BCRP] and that related to enzyme-mediated multidrug resistance pathway （GST-π and Topo Ⅱ）. RESULTS ：The IC 20 of degreasing cream ，CⅡ-3 and sorafenib were （2.40±0.16）， （200.44±27.52），（18.00±1.82）μg/mL，respectively. Compared with sensitivity group ，the protein expressions of Bcl- 2，P-gp， LRP，BCRP and Topo Ⅱ，the mRNA expressions of MDR 1， LRP，BCRP and GST-π were increased significantly in drug resistance group （P＜0.05 or P＜0.01）. Compared with @qq.com drug-resistance group ，the mRNA and protein expression of MDR1 mRNA and LRP ，BCRP，GST-π were significantly decreased in degreasing cream group and C Ⅱ-3 group（P＜ 0.05 or P＜0.01）；the protein expression of Bcl- 2 and the mRNA expression of Topo Ⅱ were significantly decreased （P＜0.01）， while the protein expression level of Cleaved-Caspase- 9 p37 was significantly increased in C Ⅱ -3 group （P＜0.05）. CONCLUSIONS：Degreasing cream and C Ⅱ-3 can reverse multidrug resistance of HepG 2/ADM cells by reducing drug efflux ， promoting cell apoptosis ，reducing the mRNA and protein expression of multi-drug resistance gene as well as gene in enzyme-mediated multi-drug resistance pathway. The effect of C Ⅱ-3 is better than that of degreasing cream.
ABSTRACT
BACKGROUND: In recent years, there have been many studies on the use of supercritical fluids for biological material treatment in countries outside China. However, little is reported on application of supercritical fluids to bone tissue extraction, in particular in China. OBJECTIVE: To evaluate the effectiveness of supercritical carbon dioxide extraction technology in the treatment of porcine femur cancellous bone and its effect on bone biological properties. METHODS: Porcine femoral bone blocks that were subjected to supercritical carbon dioxide extraction (study group) or not (control group) were prepared to determine bone mineral density, microstructure, maximum compressive strength, elastic modulus, bone tissue composition, collagen content and perform histological analysis. Bone marrow mesenchymal stem cells (BMSCs) were inoculated into two groups of bone blocks, and cultured for 1 day. The microporous structure of trabecular bone and cell adhesion and growth in bone material-cell composite were observed by scanning electron microscopy. The two groups of bone blocks were implanted subcutaneously in SD rats. The inflammatory reaction of subcutaneous tissue was observed histologically at 1, 2 and 4 weeks after surgery. The experimental protocol had been approved by the Animal Ethics Committee of Chinese PLA General Hospital, China. RESULTS AND CONCLUSION: There were no significant differences in pore size, bone mineral density, maximum compressive strength, elastic modulus and collagen content between the study and control groups (P>0.05). Scanning electron microscopy showed that in the control group, the material pores had poor connectivity and there was soft tissue residue; in the study group, material pores were connected to each other and the structure was intact. Fourier transform infrared spectroscopy and X-ray diffraction analysis showed that the two groups of bone tissue materials had similar absorption and diffraction peaks. Thermogravimetric analysis showed that supercritical carbon dioxide extraction could reduce water content in bone tissue. Hematoxylin-eosin staining showed that there were no soft tissue residues in the bone, and the cell residues in the bone pit were significantly reduced in the study group, while soft tissue and cell residues were observed in the control group. Sirius red staining and modified Masson staining showed that the structure of bone collagen in the study group was intact, the cytoplasmic components reduced, and the cytoplasmic components in the control group remained significantly. Scanning electron microscopy showed that there was no obvious cell adhesion in the control group, but cell adhesion growth was obvious in the study group. Perivascular inflammatory response in the bone tissue implantation region was obviously weaker in the study group than in the study group. These results suggest that supercritical carbon dioxide extraction technology is an effective and environment-friendly bone tissue processing technology. It can effectively remove porcine cancellous bone cells and soft tissue without affecting its collagen structure and content and mechanical properties, retaining intact bone pore structure, increasing cell adhesion and growth, and effectively reducing inflammatory rejection.
ABSTRACT
AIM To study the effects of degreasing-frosting on reducing toxicity and enhancing efficacy of Periplaneta americana L..METHODS The 95％ ethanol extracts from P.americana raw powder and processed products were extracted by petroleum ether,ethyl acetate,n-butanol and water.TLC was applied to comparing the component changes in various fractions.GC-MS was used for analyzing the kinds and contents of liposoluble constituents before and after processing.Shaking flask method combined with UV spectrophotometry method was adopted in the determination of Papp values in octanol-water and different pH values (2.23,3.28,4.22,5.23,6.12,7.24,8.23,9.10,10.15 and pure water) of buffer solutions.RESULTS After the petroleum ether fraction was processed,the colors of TLC spots became much shallower,which changed more significantly with the prolonging of processing time and increase of pressure.The TLC spots in the n-butanol and water fractions were clear without obvious changes.And no TLC spots were found in the ethyl acetate fraction.The kinds of liposoluble constituents were the same before and after processing,whose absolute contents were decreased after processing.Compared with raw powder,the Papp values of minor polypeptides in various processed products at different pH values exhibited significant differences (P ＜0.05,P ＜0.01),which also showed certain differences at the same pH values.CONCLUSION Degreasing-frosting can affect the contents of liposoluble constituents in P.americana and Papp values of minor polypeptides,which may be the mechanism of reducing toxicity and enhancing efficacy.
ABSTRACT
Introducción: la mangiferina es una glucosil xantona natural presente en varias partes de Mangifera indica L. (árbol del mango), que posee gran variedad de efectos farmacológicos. Su aislamiento, a partir de las hojas, es de vital importancia teniendo en cuenta las propiedades farmacológicas que se le atribuyen por sí sola. En Cuba, se ha desarrollado un procedimiento nacional de obtención de mangiferina a partir de hojas de M. indica, empleando un método tradicional en tanque agitado, que incluye una etapa de desengrase, sin embargo continúan los estudios encaminados al uso de métodos no convencionales de extracción como el ultrasonido, teniendo en cuenta la rapidez y eficiencia en la obtención de fitoconstituyentes. Objetivo: evaluar la etapa de extracción de la fracción apolar en el proceso de extracción de mangiferina mediante ultrasonido a partir de hojas de M. indica. Métodos: se trabajó con hojas de M. indica, molidas y almacenadas en bolsas de nylon a temperatura y humedad ambientes. Se evaluó la influencia de los parámetros de operación (tiempo y relación disolvente material vegetal) en la etapa de desengrase, mediante un diseño factorial de superficie de respuesta, utilizando el ultrasonido. Resultados: el mejor valor experimental (29,03 ± 1,10 mg/g) se alcanza para un tiempo de 90 min y una relación volumen de disolvente/ material vegetal de 47 mL/g, empleando n-hexano como disolvente. Conclusiones: el método de ultrasonido es adecuado para la extracción de la fracción apolar en el proceso de obtención de mangiferina a partir de hojas de M. indica(AU)
Introduction: mangiferin is a natural glucosyl xanthone found in several parts of Mangifera indica L. (mango tree) which has a great variety of pharmacological effects. Its isolation from leaves is of great importance, considering the pharmacological properties attributed to it. A national procedure has been developed in Cuba to obtain mangiferin from M. indica leaves using a traditional stirred tank procedure which includes a degreasing stage. However, studies continue to be conducted about the use of unconventional extraction techniques such as ultrasound, due to the speed and efficacy with which phytoconstituents may be obtained. Objective: evaluate the apolar fraction extraction stage of the process of ultrasound extraction of mangiferin from M. indica leaves. Methods: M. indica leaves were ground and stored in plastic bags at ambient temperature and humidity. Operating parameters (time and solvent / plant material ratio) were evaluated at the degreasing stage with a response surface factorial design, using ultrasound. Results: the best experimental value (29.03 ± 1.10 mg/g) is obtained for a time of 90 min and a solvent volume / plant material ratio of 47 ml/g, using n-hexane as solvent. Conclusions: the ultrasound technique is appropriate for apolar fraction extraction of mangiferin from M. indica leaves(AU)