ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
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.