ABSTRACT
Objective:Quantitative analysis of anti-inflammatory synergistic pharmacodynamics mechanism of baicalin and wogonoside by medium efficiency principle. Method:inflammatory cell model was constructed by stimulating RAW264.7 cells by lipopolysaccharide （LPS） 100 μg·L-1 in vitro. The experiment was performed in the normal group， the model group， the andrographolide group （10 μmol·L-1）， the baicalin group （2.06，4.13，8.25，16.5，33，66，132 μmol·L-1） and the wogonoside group （2.94，5.88，11.75，23.5，47，94，188 μmol·L-1） and the baicalin-wogonoside combination group ［（2.06+2.94）（4.13+5.88）（8.25+11.75）（16.5+23.5）（33+47）（66+94）（132+188） μmol·L-1］. The levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in the cell culture supernatants after drug intervention for 50 min and 4 h were detected by enzyme-linked immunosorbent assay （ELISA） method. The level of nitric oxide （NO） in the cell culture supernatant after drug intervention for 24 h were detected by Griess method. Western blot was used to detect the activation levels of phosphorylation of nuclear factor-κB p65（p-NF-κB p65） and inducible nitric oxide synthase（iNOS） in cells after drug intervention for 2 h and 12 h. The fa/fu-dose profile of each indicator was drawn to observe the increase or decrease of effect. Result:Compared with normal group， the expression of p-NF-кB p65， iNOS and cytokines including TNF-α， IL-6 and NO （P<0.05，P<0.01） in the model group were significantly up-regulated. Compared with the model group， each group at high doses could inhibit the phosphorylation of NF-кB p65 protein（P<0.05），the baicalin group and the combined group could down-regulate the expression of iNOS protein in a concentration-dependent manner（P<0.01） and the baicalin group had no obvious inhibitory effect. each administration group at high dose could significantly inhibit the production of NO（P<0.05），but each group had no inhibitory effect on IL-6 production. The baicalin group and the combined group could significantly Inhibit the production of TNF-α（P<0.05） and there was no significant difference between the baicalin group and the model group. At the experimental dose， the fa/fu-dose table showed that the fa/fu value of p-NF-кB p65 and IL-6 in the combined group was not greater than the baicalin group and the wogonoside group. The fa/fu value of iNOS， TNF-α and NO in the combined group is higher than the baicalin group and the wogonoside group. Conclusion:The baicalin and wogonoside have different effects on different targets in the NF-κB pathway. The wogonoside is the main pharmacological substance in this combination and the combination shows different degrees of synergy or antagonism effects on different targets.
ABSTRACT
Objective:To observe effect of long-term administration of rhein on the kidney toxicity of mice, and explore its possible toxic mechanism, in order to provide some basis for rational clinical drug use and further research. Method:The 30 Kunming mice (half male and half female) were randomly divided into 3 groups:control group, low-dose rhein group and high-dose rhein group (0.175,0.35 g·kg-1), with 10 mice in each group. The intragastric administration lasted for 60 days. During administration, general situations of the mice were observed and recorded. Serum urea nitrogen (BUN), creatinine (SCr), malondialdehyde (MDA), superoxide dismutase (SOD), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were detected after drug withdrawal. Kidney index was calculated, and glutathione peroxidase (GSH-Px) and reduced glutathione/oxidized glutathione (GSH/GSSG) ratio were measured. The kidneys were collected and histopathologically examined, and the protein expressions of transforming growth factor beta (TGF-β1) and cysteine aspartic acid specific protease-3 (Caspase-3) were detected by immunohistochemistry. Result:Compared with the control group of the same sex, BUN and SCr of the administration group increased significantly(PPPPα and Caspase-3 increased significantly(PPPPβ1 was increased(PConclusion:The toxicity of rhein in the kidney of mice was obvious at the dose of 0.35 g·kg-1·d-1, and the toxicity in male organism is more obvious. The mechanism of its potential toxicity may cause the imbalance of glutathione antioxidant system, induce excessive oxidation, trigger inflammatory reaction, activate the expression of Caspase-3, and then induce apoptosis.