| Xinfeng Capsule, the clinical experience prescription of the state administration of traditional Chinese medicine of The First Affiliated Hospital of Anhui University of traditional Chinese medicine(approval number: anhui words Z20050062 medicine system). Clinical studies have identified that Xinfeng Capsule has unique advantages and less side effects as one of the ideal Traditional Chinese Medicine(TCM) on rheumatoid arthritis therapy. Xinfeng Capsule is traditional Chinese medicine theory as the guiding principle, dialectical therapy from the overall macro conditions, and the subjective wishes. The study is to investigate the role of formula ingredients and the curative effect of different dosage ratio formula from the objective aspects, thus select the best compatibility formula, then to study the main composition change of prescription before and after optimization from the perspective of material base. This study could provide theoretical basis for clinical application and the foundation of scientific formula.Objective:1. Select the best compatibility prescription of Xinfeng Capsule to treat of rheumatoid arthritis, reveal the compatibility prescription rule;2. Evaluate and validate the best compatibility prescription of Xinfeng Capsule by the pharmacodynamics experiment;3. Investigate material basis change of the prescription before and after optimization of Xinfeng Capsule.Methods:1. According to the uniform design plan U6×(64) applied, four materials of traditional Chinese medicine in the prescription of Xinfeng Capsule the as flavor factor study, each flavor factor has six levels, obtained the prescription 1-6;2. Establish a rat model of AA, grouping, each group of rat gavages the appropriate prescription;3. Observe the survival status, weight changes, and toes swelling of each group of rat;4. Collect and prepare the specimens of rat blood and ankle by modern pharmacological methods;5. Determine content of each group rat serum cytokines IL-1β, IL-6, IL-17, IL-10,TGF-β1 by ELISA method; 6. Verify the best compatibility prescription of Xinfeng Capsule, which determines content of JNK/p-JNK of each group rat ankle by Western blot method;7. Investigate material basis change of the prescription before and after optimization of Xinfeng Capsule by TLC and HPLC method.Results:1. Select the best compatibility prescription of Xinfeng Capsule:(1) Rats general condition: Normal control group rats color shinning, normal limbs activities; Model control group rats appeared dispirited, multiple joint swelling and activities outside obstacles, etc.; among goup 1 to group 6, group 3, group 4, group 6best anti-inflammatory effect; the status of group 1, group 5 similar with the model group;other groups had no significant difference(2) Changes in rats paw edema secondary degree: group 2, group 3, group 4, group 6 rat paw swelling degree of secondary significantly lower than those in the control group;group 3, 4, 6 best anti-inflammatory effect; group 2,5 in the early period of better treatment of inflammation, later poor; group 1 no ideal treatment.(3) From the experimental results, IL-1β, IL-6, IL-17 evaluation of efficacy, by multiple regression analysis and synthesis rates combination, obtain optimized prescription:Astragalus 28.293 g, Myotonin 16.984 g, Tripterygium Wilfordii 11.800 g, Centipede0.970 g(change Adult-dosage everyday: Astragalus 4.491 g, Myotonin 2.696 g,Tripterygium Wilfordii 1.873 g, Centipede 0.154 g).2. Optimization prescription pharmacodynamic validation study results:(1) Compared with normal group, serum cytokine levels of IL-10, TGF-β1 reduced in model group rats, however serum cytokine levels of IL-10, TGF-β1 elevated in Xinfeng Capsule group and optimization prescription group, and that the effect of prescription group was better than Xinfeng Capsule group.(2) In this experimental, Western-blotting assay Xinfeng Capsule and synovial tissue optimization side AA rats JNK, p-JNK affect protein expression, result indicated that,Compared with normal group, synovium in JNK, p-JNK protein expression of model group rats were significantly increased; Compared with model group, synovium in JNK,p-JNK protein expression of Xinfeng Capsule group, optimization prescription group and positive control group were subject to different degrees of inhibition; Compared with Xinfeng Capsule group, optimization prescription regulating synovium AA abnormal expression of JNK, p-JNK protein is more significant.3. A preliminary study of main composition change of Xinfeng Capsule prescription before and after optimization:(1) Result of qualitative identification of Xinfeng Capsule prescription before and after optimization: no negative interference of optimization prescription. Spot 1(fluorescence), spot 2(brown) had the corresponding position in chromatogram of optimization prescription and astragalus, Xinfeng Capsule had just spot 1(fluorescence), which showed that the chromatogram of optimization prescription and astragalus chromatography corresponding spots increase may be due to the scientific and reasonable compatibility and dose relations has increased the interaction of various ingredients composition of traditional Chinese medicine compound, dissolution of effective components in the compound.(2) The result of quantitative study of Xinfeng Capsule prescription before and after optimization: The retention time of calycosin glucosidase in Xinfeng Capsule and optimization prescription were consistent with calycosin glucoside reference substance,and negative non-interference. Content determination results show that the content of calycosin glucoside in Xinfeng Capsule and optimization prescription had no significant difference.Conclusion:Xinfeng Capsule was optimized by the uniform design optimized formula for optimization prescription, Astragalus 28.293 g, Myotonin 16.984 g, Tripterygium Wilfordii 11.800 g, Centipede 0.970 g(change Adult-dosage everyday: Astragalus 4.491 g, Myotonin 2.696 g, Tripterygium Wilfordii 1.873 g, Centipede 0.154 g). The pharmacodynamics experiment research of optimization prescription shows that,optimization prescription was better than Xinfeng Capsule in terms of anti-inflammatory immune regulation, and optimization prescription could reduce the dosage of the formula of the original medicinal materials. A preliminary study of main composition change of Xinfeng Capsule prescription before and after optimization showed the effective ingredient dissolution increased by formula optimization, and the content of calycosin glucoside in astragalus had no significant change before and after optimization. |
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