| Objective: To analysis the transitional ingredient of rats serum with Collageninduced arthritis(CIA) by the technology of liquid chromatography-mass spectrometry(LC-MS) after these rats are fed with Guizhi, Fuzi,and the combination with Guizhi and Fuzi warm meridians separately, to observe the influence that modeling may affect to the serum contained with drug, and to further investigate what the basis material after fed with combination with Guizhi and Fuzi transfer into the blood and their interaction under the environment of compatibility, that aim to find out the theoretical basis to support the view of that the combination with Guizhi and Fuzi warm meridians compatibility is able to treat rheumatoid arthritis(RA).Methods: There are two stages to carry out the experiment inclusive of preparation for serum contained with drug and investigation on what the basis material is after rats fed with the combination Guizhi and Fuzi transfer into the blood.The first stage prepare 35 Wistar male rats and divide into five groups randomly such as the Normal Group, the Single-Dose Group, the Five TimesDose Group, the Ten Times-Dose Group and the Twenty Times-Dose Group.There are 7 rats in each group to be fed with the combination Guizhi and Fuzi by dose of single time, five times, ten times and twenty times respectively except for that the rats from the Normal Group are fed with normal saline. Take blood from the rats through their hepatic portal vein after fed 15 min, 30 min, 60 min, 90 min, 120 min, 180 min, 240 min, and deal with serum by the method of methanol precipitation, acetonitrile precipitation, trichloroacetic acid precipitation and ethyl acetate precipitation, then analysis them by high performance liquid chromatography(HPLC). Thus, the way of getting peaks in well-separated, high sensitivity and maximum detected, will be found out to be the final plan for preparation of the serum contained with drug.The Second stage prepare 35 Wistar male rats. Pick out 6 rats randomly and put into the Normal group and the combination with Guizhi and Fuzi group, each group has 3 rats, while the remained rats are ready to be modeled with type Ⅱ collagen. Those rats are succeeded to be modeled, were randomly divided into five groups by average,the model group, the model of fed with Guizhi group, the model of fed with Fuzi group, the model of fed with combination with Guizhi and Fuzi group. Feed the rats with 20 times drug dose;the rats from the normal group and the model group are fed with normal saline.Take the rats’ blood via the hepatic portal vein after fed 60 minutes, and dealt with serum in methanol precipitation method,and analysed by the technology of liquid chromatography-mass spectrometry(LC-MS). Comparing with spectrograms from the serum contained with drug group, the normal group, the model group, the test solution and the reference sample in the same conditions, analysis the transfer ingredient in the blood after rats fed with drugs, investigate what the material basis after fed with combination with Guizhi and Fuzi transfer into the blood and their interaction under the environment of compatibility,establish the blood fingerprints for Guizhi, Fuzi, and combination with Guizhi and Fuzi warm meridians compatibility.Results:(1)There are 10 kinds of transitional ingredients in the combination with Guizhi and Fuzi group’serum contained with drug, all of them are metabolites.(2)There are 10 kinds of transitional ingredients in the model of fed with Guizhi group’ serum contained with drug, one is prototype component an d 9 of them are metabolites.(3)There are 9 kinds of transitional ingredients in the model of fed with Fuzi group’ serum contained with drug,2 of them are prototype components and 7 of them are metabolites.(4)There are 25 kinds of transitional ingredients in the model of fed with combination with Guizhi and Fuzi group’ serum contained with drug, 8 of them are prototype components and 17 of them are metabolites.Conclusion:(1)Study shows that the numbers of transition ingredients in drug contained serum are obviously increased after modeling, and the relevant in gredients concentration are higher than non-model’s. Thus, the type Ⅱ collagen-induced autoimmune arthritis has a certain effect on the preparation of drugcontained serum.(2)There are 25 kinds of derived ingredients in the model of fed with combination with Guizhi and Fuzi group’ serum contained with drug, 8 of them are prototype components and 17 of them are metabolites.These compounds may become the material basis for the combination with Guizhi and Fuzi warm meridians compatibility.(3)Transition ingredients in serum contained drug with combination with Guizhi and Fuzi are more than the total quantities of serum with sole Guizhi plus Fuzi. Besides,some of the ingredients can be absorbed by the organism only in the conditions of combination with Guizhi and Fuzi, and prompt the organism absorbing the effective materials.It is becoming the theoretical basis of analysis for the transition ingredients in the serum with combination with Guizhi and Fuzi warm meridians compatibility, and provides a method for investigation on the regularity of prescription compatibility. |
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