| Objective: The content of cimicin,5-o-methylvisamycin and the content of5-o-methylvisamycin in Fangfeng powder were determined by different decocting time,the effect of different decocting time on the pharmacodynamics of Fangfeng powder was studied by replicating cold animal model,to explore the best decocting time of Fangfeng,to provide experimental basis for clinical application of special Decocting Methods of traditional Chinese medicine.Methods: High performance liquid chromatography(HPLC)was used to determine the content of cimicin and 5-o-methylvisamifoside in Fangfeng single medicine and Fangfeng compound Fangfeng powder.Chromatographic column:INERTSIL ODS-3(250×4.6 mm,5?m),Mobile phase:Methanol-Water(40 : 60),Column temperature:30?,Current Speed:1 mL·min-1,Detection wavelength:254 nm,Injection volume:10 ?L.Hot plate method,yeast induced fever method and plantar swelling method were used to observe the effects of fangfengsan Decoction at different times on the analgesic,antipyretic and anti-inflammatory effects of model animals.Detection of immunoglobulin IgA,IgM and IgG in the serum of rats with swelling foot by ELISA.Western blot was used to detect the protein expression of Toll receptor 2(TLR2),Toll receptor 4(TLR4)and myeloid differentiation factor 88(my D88)in spleen tissue of rats in each group.Results: When Decoction of Fangfeng pieces for 30 minutes,the content of cimicin is7.547 mg·g-1,the highest content,the content of 5-o-methylvisamycin was 3.472 mg·g-1 in25-35 min of decocting,which was the highest and basically stable.The total content of cimicin and 5-o-methylvisamycin in Fangfeng powder was 7.547 mg · g-1 when it was decocted for 10 minutes first.The results of analgesia experiment showed that,Compared with the blank control group(BCG),ibuprofen group,ganmaoshufeng tablet group andFangfeng Decoction group increased significantly in 10 min and 15min(P<0.01).The experimental results show that,compared with BCG,the body temperature of model group(mg)rats increased at all time points after modeling(P<0.05 or P<0.01);Compared with Mg,the temperature of mice in ibuprofen group was significantly lower than that in Fangfeng powder group(0 min,5 min,10 min,15 min).The results of anti-inflammatory experiments show that,compared with BCG,the swelling rate of toes in mg rats increased significantly at all time points(P<0.01);Compared with Mg,the swelling rate of toes in ibuprofen group and Fangfeng Decoction group decreased significantly(P<0.01).The results of ELISA showed that the content of IgA,IgG and IgM of rats in fangfengsan group was significantly higher than that of mg group(P<0.01 or P<0.05),Among them,the effect of decoction of Fangfeng for 10 min is the most significant(P<0.01).The results of Western blot showed that the contents of TLR2,TLR4 and MyD88 in spleen tissue of Mg rats were significantly higher than that of BCG;Compared with Mg,the contents of TLR2,TLR4 and MyD88 protein in spleen tissue of rats in fangfengsan group were decreased in different time,especially in fangfengxian Decoction group for 10 minutes(P<0.01).Conclusion: The total contents of cimicin and 5-o-methylvisamycin were the highest when the radix sileris single drug was decocted for 10 min.In the compound radix sileris powder,the total content of active ingredients was the highest when it was first decocted for10 min,and the antipyretic,analgesic and anti-inflammatory effects of radix sileris powder on the syndrome of wind cold exterior were significant when it was decocted for 10 min,and the most significant regulatory effects on inflammatory cytokines and inflammatory proteins in TLRs / MyD88 signal pathway were found. |
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