lipid-a and baicalin

HuanglianJiedu decoction (HJD) is a classic prescription for heat-clearing away and detoxifying, which is used for the clinical treatment of sepsis, due to sepsis refers to the systemic inflammatory response induced by infection in western medicine, and infection belongs to the category of poison-heat syndrome in traditional Chinese medicine.. Previous study had elucidated the effective components from HJD with high affinity to lipid A, which can generate the release of pro-inflammatory-cytokines, resulting in sepsis. Now the anti-sepsis activities of these compounds were evaluated.. Immunofluorescence, immunohistochemical staining, ELISA and MTT methods were used to evaluated these compounds.. Immunofluorescence analysis evaluated the effects of compounds on the binding of FITC-LPS to RAW264.7 cells, and showed the fluorescence intensity was significant attenuated in geniposides, palmatine, baicalin and berberine groups (64 and 128 μg/mL) compared with model group (p < 0.05), which showed these compounds inhibit the combination of LPS with receptor of cells; immunohistochemical staining and ELISA method showed the TLR4 receptor expression, IL-6 and TNF-α levels were significant decreased in the groups treated with compounds, indicating that geniposides, baicalin, palmatine and berberine can play the role of anti-sepsis by inhibiting the expression of TLR4, the releasing of IL-6 and TNF-α; MTT assay showed that palmatine and berberine had a weak effect on cell viability, while others not, indicating that the compounds have protective activity.. It could be concluded the high affinity binding between these compounds and lipid A may be an important basis for its anti-LPS activity in vitro.

Topics: Animals; Berberine; Berberine Alkaloids; Cell Line; Drugs, Chinese Herbal; Flavonoids; Interleukin-6; Iridoids; Lipid A; Lipopolysaccharides; Mice; RAW 264.7 Cells; Sepsis; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha

Huanglian Jiedu Decoction (HJD), the classical recipe for relieving fever and toxicity, has been used for treating sepsis in China for sixteen years. However, the effective components of HJD have not been elucidated until now. Therefore, there is a need to elucidate the effective components of HJD against sepsis on animal models induced by endotoxin (LPS). The affinity force of the effective components of HJD with lipid A was evaluated by a biosensor.. Lipid A is regarded as the bioactive center of LPS and is always used as a drug target. In order to obtain the effective components of HJD against sepsis, seven fractions from HJD were tested by a biosensor method for assessing the affinity for lipid A. After further separation, the components were isolated from high lipid A-binding fractions and their affinities to lipid A were assessed with the aid of a biosensor. Their activities were then assayed by an in vivo experiment administered through a tail vein injection. The levels of LPS, TNF-α, and IL-6 from the blood were found and pathology experiments were performed.. Three out of the seven fractions exhibited high lipid A-binding affinities. Berberine, baicalin and geniposide were obtained from the three high lipid A-binding fractions. The animal experiments indicated that the levels of LPS, TNF-α and IL-6 in the medicated treatment groups were much lower than that of the model group ((**)P<0.01). The medicated treatment groups exhibited stronger protective activities on varying organs in the animal model.. Berberine, baicalin and geniposide could neutralize LPS by binding with lipid A and then reduce the release of IL-6 and TNF-α induced by LPS. Furthermore, berberine, baicalin and geniposide exhibited protective activities on varying organs compared to the animal model established by the LPS-induced. These results validate that the components from HJD neutralized LPS and then depressed the release of IL-6 and TNF-α induced by LPS. This gives further evidence that HJD would be a suitable treatment for sepsis and protecting vital organs.

Topics: Animals; Berberine; Biosensing Techniques; Drugs, Chinese Herbal; Female; Flavonoids; Interleukin-6; Iridoids; Kidney; Lipid A; Lipopolysaccharides; Liver; Lung; Male; Mice, Inbred BALB C; Myocardium; Sepsis; Tumor Necrosis Factor-alpha