Beneficial effects of Houttuynia cordata polysaccharides on “two-hit” acute lung injury and endotoxic fever in rats associated with anti-complementary activities

Houttuynia cordata Thunb. is a traditional herb used for clearing heat and eliminating toxins,and has also been used for the treatment of severe acute respiratory syndrome(SARS). In vitro, the crude H. cordata polysaccharides(CHCP) exhibited potent anti-complementary activity through both the classical and alternative pathways by acting on components C3 and C4 of the complement system without interfering with the coagulation system. This study was to investigate the preventive effects of CHCP on acute lung injury(ALI) induced by hemorrhagic shock plus lipopolysaccharide(LPS) instillation(two-hit)and LPS-induced fever in rats. CHCP significantly attenuated pulmonary injury in the "two-hit" ALI model by reducing pulmonary edema and protein exudation in bronchoalveolar lavage fluid(BALF). In addition, it reduced the deposit of complement activation products in the lung and improved oxidantantioxidant imbalance. Moreover, CHCP administration inhibited fever in rats, reduced the number of leukocytes and restored serum complement levels. The inhibition on the inappropriate activation of complement system by CHCP may play an important role in its beneficial effects on inflammatory diseases. The anti-complementary polysaccharides are likely to be among the key substances for the heatclearing function of H. cordata.

Simultaneous determination of six steroidal saponins and one ecdysone in Asparagus filicinus using high performance liquid chromatography coupled with evaporative light scattering detection

A high-performance liquid chromatography coupled with an evaporative light scattering detector(HPLC-ELSD)has been developed to evaluate the quality of Asparagus filicinus through a simultaneous determination of six steroidal saponins and one ecdysone,including aspafiliosides A,B,C,E,G,filiasparoside A and 20-hydroxyecdysone.With a C_(18) analytical column,the seven analytes were separated efficiently using acetonitrile-water as the mobile phase in a gradient program.The method limits of detection ranged 0.1250.225μg,and the method limits of quantitation ranged 0.408-0.720μg,respectively.The intra-and inter-day precisions of the method were evaluated and were all less than 3%.All the recoveries for the spiked analytes ranged 95.16%-100.61%.The proposed method was succesfully applied to quantify the seven components in thirteen samples from different localities in China.

Development of a C3c-based ELISA method for the determination of anti-complementary potency of Bupleurum polysaccharides

Traditionally, determination of inhibitory potency of complement inhibitors is performed by the hemolytic assay. However, this assay is not applicable to the lectin pathway, thus impeding the understanding of complement inhibitors against the overall function of the complement system. The main objective of our study was to develop a specific enzyme-linked immunosorbent assay(ELISA) as an alternative method to assess the anti-complement activity, particularly against the lectin pathway. By using respective coating substrates against different activation pathways, followed by capturing the stable C3c fragments, our ELISA method can be used to screen complement inhibitors against the classical pathway and the lectin pathway. The inhibitory effect of suramin on the classical pathway, as measured by our hemolytic assay is consistent with previous reports. Further assessment of suramin and Bupleurum polysaccharides against the lectin pathway showed a good reproducibility of the method. Comparison of the lectin pathway IC50 between Bupleurum smithii var. parvifolium polysaccharides(1.055 mg/mL) and Bupleurum chinense polysaccharides(0.98 mg/mL) showed that, similar to the classical and alterative pathway, these two Bupleurum polysaccharides had comparable anti-complementary properties against the lectin pathway. The results demonstrate that the described ELISA assay can compensate for the shortcomings of the hemolytic assay in lectin pathway.

ISOLATION, CHARACTERIZATION AND MEDICAL PROPERTIES OF ANTICOMPLEMENT AGENTS FROM TRADITIONAL CHINESE MEDICINES

The inappropriate activation of complement system may cause some life-threatening symptoms such as rheumatoid arthritis,systemic lupus erythematosus(SLE)and acute respiratory distress syndrome(ARDS).In our efforts to obtain natural anticomplement agents from traditional Chinese medicines(TCMs)for prevention and treatment of the complement-associated diseases。