Study on components with anti-complement activity from ethanol extract of Sanse tablets by UPLC-Q-TOF-MS/MS / 药学实践杂志

Objective To analyze the chemical components in the ethanol extract of Sanse tablets with anti-complement activity, and to provide scientific basis for its therapeutic effects. Methods The classical anti-complement pathway was used to compare the activity of different portions of Sanse tablet alcohol extract, and to identify the fraction with anti-complement activity. The chemical composition in active fraction was analyzed by UPLC-Q-TOF-MS/MS. The chemical components were identified by comparing the retention time, exact molecular weight and mass spectrum information with the standard chemicals. Results The ethyl acetate fraction of the Sanse tablet ethanol extract had the best anti-complement activity. 42 chemicals were identified, including 16 alkaloids, 15 terpenoids, 6 flavonoids and 5 phenolic acids. Conclusion The characterization of the chemical components in the anti-complement active fraction of Sanse tablets provides a scientific basis for the therapeutic effects of Sanse tablets, which will help the future development of the compound preparations of Chinese medicine in our hospital.

Structural characterization and anti-complement activity of polysaccharides from Glechomae Herba / 中草药

Objective: To isolate and purify the homogeneous polysaccharides from Glechomae Herba through the guidance of anti-complement activity, and study their structures. Methods: Crude polysaccharides from Glechomae Herba were extracted by water extraction and alcohol precipitation, then homogeneous polysaccharides were separated by DEAE-52 and Sephadex G-100 column chromatography. The structures of homogeneous polysaccharides were characterized by HPGPC, IR, GC, methylation and NMR, and their anti-complement activities were also determined. Results: Two homogeneous polysaccharides GLP-1 and GLP-2 were obtained with the molecular weight of 5 370 and 20 040. They were both heteropolysaccharides composed of arabinose and galactose with the ratio of 1:6.3 and 1:4.9, respectively. Conclusion: The structures of GLP-1 and GLP-2 further elucidate the pharmacodynamic basis of their anti-complement activities and provide basis for screening natural complement inhibitors.

Role of polyphenols in the antimicrobial activity of ethanol Tamarindus indica L leaves fluid extract / Papel de los polifenoles en la actividad antimicrobiana del extracto fluido de las hojas de Tamarindus indica L

The aim of this work was to explore in an active, fractioned, and chemically characterized Tamarindus indica L. (TIL) leaves extract, the influence of flavonoids and polyphenol compounds on the antimicrobial activity. A spectrophotometric quantification of the total phenols and flavonoids content was determinate to the TIL leaves extract, as well as, to the four fractions in which was fractioned (n-hexane, chloroform, ethyl acetate and n-butanol). The extracts and their fractions were microbiologically tested against six ATCC bacteria and Candida albicans, being determined their minimum inhibitory and bactericidal concentrations (MIC and MBC). Additionally, the extracts were evaluated in their influence on human complement system (classical and alternative pathways). Fractions with high content of flavonoids and polyphenols (ethyl acetate and n-butanol) are active against Bacillus subtilis and inhibit the human complement system (direct pathway, IC50 31.05 and 33.65 ug/mL respectively), but are not active over Staphylococcus aureus. However, this bacterium was susceptible to fractions with low or null concentration of flavonoid or polyphenol compounds. No fractions neither the fluid extract were active against Salmonella typhimurium and Candida albicans. Experimental data suggest that phenols and flavonoids are not the only components involved in the antimicrobial activity of TIL leaves as has been previously suggested by other authors. Complement activity tests did not support a putative role on the antimicrobial activity.

El objetivo de este trabajo fue explorar en un extracto activo de hojas de Tamarindus indica L. (TIL), fraccionado y caracterizado químicamente, la influencia de los polifenoles y flavonoides en su actividad antimicrobiana. Se cuantificaron por espectroscopia UV-visible los contenidos de fenoles totales y flavonoides en el extracto de TIL así como de las cuatro fracciones obtenidas (n-hexano, cloroformo, acetato de etilo y n-butanol). Se evaluó la actividad microbiológica del extracto y sus fracciones contra seis bacterias ATCC y Candida albicans, determinándose sus concentraciones mínimas inhibitorias y bactericidas (MIC y MBC). Adicionalmente, se evaluó la influencia de los extractos en el sistema de complemento humano (vía clásica y alternativa). Las fracciones con altas concentraciones de polifenoles y flavonoides (acetato de etilo y n-butanol) fueron activas contra el Bacillus subtilis e inhibieron el sistema de complemento humano (vía directa, IC50 31.05 y 33.65 g/mL, respectivamente), pero no fueron activas contra Staphylococcus aureus. Sin embargo, esta bacteria fue susceptible a fracciones con baja o nula concentración de polifenoles y flavonoides. El extracto fluido y todas sus fracciones resultaron inactivos frente a Salmonella typhimurium y Candida albicans. Los datos experimentales sugieren que los fenoles y flavonoides no son los únicos compuestos involucrados en la actividad antimicrobiana de hojas de TIL, como había sugerido por otros autores. La actividad medida sobre el sistema de complemento, no aporta relevancia a la actividad antimicrobiana de las hojas de TIL.

Suero de conejo con actividad de complemento para tipificación del complejo mayor de histocompatibilidad humano / Rabbit serum with complement activity to typify human major histocompatibility complex

En el ensayo de microlinfocitotoxicidad, el suero de conejo como fuente de complemento desempeña un rol esencial en la clasificación del complejo mayor de histocompatibilidad humano, tanto para la tipificación antigénica como en el cross-match linfocitario de la pareja donante-receptor antes de realizar los trasplantes. En este trabajo, se obtuvieron 3 lotes experimentales de dicho hemoderivado con óptimos indicadores de rendimiento, actividad biológica y estabilidad. El exanguíneo de los animales se realizó por la técnica de yugulación. El suero fue separado por centrifugación en condiciones ambientales y de temperatura controladas. Posteriormente, el producto se sometió a filtración esterilizante y se tomaron muestras para los ensayos de calidad. El estudio mostró que el producto conserva la actividad biológica al menos durante 18 meses y que los valores de citotoxicidad se encuentran dentro de los límites de aceptación para su empleo en los ensayos de histocompatibilidad pretrasplante

Microlinfocitotoxicity assay is one of the serological methods used to classify human major histocompatibility complex. In this technique the rabbit´s serum as complement source, plays an essential role in antigens determination and in lymphocytes cross match assay as necessary stage before the selection of the best donor-receiver pair to realize the organ transplant. Three experimental lots of normal rabbit serum were developed with excellent indicators of efficiency, biological activity and stability. Rabbit's blood was obtained through jugulating technique and serum was separated by centrifugation in controlled environment and temperature´s conditions. The sterilization process was performed by filtration and the samples were taken for quality´s control assays. Stability studies showed that the product kept the biological activity at least during 18 months after it was processed and cytotoxicity's values were adequate for its employment in histocompatibility pre-transplant assays

The study on expression and activity of human mutant CD59 gene / 中国免疫学杂志

Objective: To amplify two human mutant CD59 eukaryotic expressing systems and investigate mutant CD59 functional activity. Methods: Mammalian expression vector PLATER of mutant CD59 cDNAs was transfected into CHO together with the pcDNA by lipofectamine,which confered resistance to G418(400 ?g/ml). The positive clones were tested by FIH. Activity of both mutants CD59 was determined by BCECF release assay. Results: Mutant CD59 cDNAs subcloned into the mammalian expression vector PLATER and transfected CHO together with the pcDNA,which confered resistance to G418. The positive clones were tested by FIH.Activity of both mutants CD59 before and after glycation was determined by BCECF release assay,both of them could restrict MAC formation ,and glycation could inhibit CD59. Conclusion: A eukaryotic system that expressing mutant CD59 cDNA was successfully set up.It was found that mutant CD59 could restrict MAC formation,and glycation could inhibit mutant CD59. These would be helpful for the furthur study of link mutant CD59 and the vascular proliferative of diabetes.