Effects Of Different Decoction Times On The Active Ingredients And Medicinal Effects Of Chaihu And Wuchaihu Drink

Objective:To observe the effect of different decoction time of Bupleurum and its compound Wuchaihuyin,the content of the effective components of Saikosaponin a and Saikosaponin d was tested,and the optimal decoction time of Bupleurum was investigated.Yeast-heated rat model,foot swelling rat model,explore the antipyretic,analgesic,and anti-inflammatory effects of Chaihu decoction at different decoction times,and explore the impact on the immune response mediated and TLRs/NF-κB signaling pathway by Wu Chai Hu Yin.Methods:According to Pharmacopoeia requirementsUsing Hplc,using Hplc to test the contents of saikosaponin a and saikosaponin d in Bupleurum and Wuchaihuyin with different decoction time.Use 15%yeast suspension heating,hot plate test,acute inflammation caused by egg white to observe the effects of cold-related model animal body temperature,pain threshold,toe swelling rate,spleen coefficient,thymus coefficient,immune factors（IL-4,IL-10,TNF-αand TLR-4）levels by Wuchaihuyin with different decoction time.Results:The result of Hplc shows that saikosaponin a and saikosaponin d had a good linear relationship with the peak area within a certain concentration range（Ra2=0.9993,Rb2=0.9993）.When the Bupleurum is boiling for 35 minutes,that is Bupleurum is first boiling for 15 minutes,the total content of saikosaponin a and saikosaponin d is at the highest,the contents were 34.92 mg·g^-1 and 34.25 mg·g^-1.Body temperature test results show that,compared with the blank control group（BCG）,the body temperature of the rats in the model group（MG）are significantly increased at various time points after 2.5 h（P<0.05 or P<0.01）;Compared with MG,the body temperature of rats are decreased significantly at various time points after 2.5 h in the group of Ibuprofen and the group of Bupleurum first boiling 10 min,15 min and 20 min（P<0.05 or P<0.01）.The result of the pain threshold test shows that compared with the BCG,the pain threshold of mice is significantly increased in the group of cold dispelling wind tablet,the group of Ibuprofen and the group of Bupleurum first boiling10 min,20 min and 25 min（P<0.05 or P<0.01）.The foot swelling rate test results show that,compared with BCG,the foot swelling rate of MG rats was significantly increased after modeling（P<0.01）;Compared with MG,the foot swelling rate of rats in the group of Ibuprofen and the group of Bupleurum first boiling for 0 min,10 min,15 min,and 20 min（P<0.05 or P<0.01）.The spleen and thymus coefficient test results show that,compared with BCG,the spleen and thymus coefficients of MG animals increased significantly（P<0.01）.Compared with MG,the spleen and thymus coefficients of rats in each administration group were lower than MG.ELISA test showed that compared with the BCG,the serum levels of IL-4,IL-10,TNF-αand TLR-4 in the model group were significantly increased（P<0.01）.Compared with the MG,the levels of IL-4,IL-10,TNF-αand TLR-4 were significantly reduced in Ibuprofen group and Bupleurum first boiling for 10 min,15 min,and 20 min group（P<0.01）.Western Blot test results showed that compared with the BCG,the expression levels of IL-4,IL-10,TNF-αand TLR-4 in the spleen of the model group were significantly increased（P<0.01）.Compared with MG,the levels of IL-4,IL-10,TNF-αand TLR-4 in the spleen tissue of rats in each administration group were significantly reduced（P<0.05 or P<0.01）.Conclusion:During the decoction process,the content of saikosaponin a and saikosaponin d in the decoction solution is highest when the Bupleurum first boiling for 15min.Its antipyretic,analgesic and anti-inflammatory effects are the most significant.Therefore,Bupleurum should use a special decoction method（ie,"first boiling"）in the solution of the formula,it can achieve the maximum dissolution of effective ingredients,and improve clinical efficacy.