| Tripterygium wilfordii is the dried root of tripterygium wilfordii Hook.f.,which is one of the most reliable traditional Chinese medicines for immunosuppression.Tripterygium wilfordii preparation is a traditional Chinese medicine preparation made by extracting and purifying the liposoluble mixture of tripterygium wilfordii.The products currently on the market include tripterygium wilfordii tablets,tripterygium total terpenes tablets,tripterygium polyglycoside tablets,it has a variety of immunosuppressive and non-specific anti-inflammatory effects,clinically used to treat rheumatoid arthritis,systemic lupus erythematosus,and nephrotic syndrome with definite effect.In recent years,tripterygium wilfordii preparation has attracted much attention in the treatment of various primary and secondary nephritis,such as the treatment of chronic glomerulonephritis,Henoch-Schonlein purpura nephritis(HSPN),Lupus nephritis(LN)and other diseases,tripterygium wilfordii preparation can significantly improve the prognosis of patients with various kidney diseases,however,it also showed significant toxicity,of which kidney is one of the main toxic target organs.At present,the material basis of renal toxicity caused by tripterygium wilfordii preparations is unclear,lack of scientific and effective quality control indicators and methods,as a result,the content of main toxic components cannot be controlled in the production process,which affects the quality and safety of the preparation.Therefore,how to reduce or control the toxicity of tripterygium wilfordii preparation and develop a safe and effective tripterygium wilfordii preparation have a broad development prospect.In this paper,serum pharmacology and serum medicinal chemistry studies on tripterygium wilfordii polyglycoside tablets,in order to further elaborate the material basis of nephrotoxicity of Tripterygium wilfordii polyglycoside.The thesis is divided into four chapters,the main contents are as follows:Chapter 1 Establishment of fingerprint of Tripterygium wilfordii polyglycosides tabletsIn this study,three different batches of Tripterygium wilfordii polyglycosides tablets were analyzed by HPLC,and the similarity evaluation is carried out through "the similarity evaluation system of chromatographic fingerprint of traditional Chinese medicine version A",19 common peaks were identified.The results of methodological investigation show that the precision of the instrument and the reproducibility of the experimental method are good,the similarity of the fingerprints of the three batches of samples is 0.94,which is high,indicating that the content of tripterygium wilfordii polyglycoside tablets in different batches is stable and controllable,which provided the basis for the next step of pharmacological experiment to supply qualified drugs.Chapter 2 Serum pharmacology of Tripterygium wilfordii polyglycoside tabletsUsing serum pharmacological method,the effects of different rat blank serum and DMSO dosage on cell activity were investigated,and the dosage of oral administration of experimental animals and the way of adding serum to the culture system were studied,finally,it was established that the extract of Tripterygium wilfordii polyglycoside tablets(100 times of clinical equivalent dose)was given to beagle dogs by intragastric administration and collect blood,protein precipitated by acetonitrile in serum containing drugs at different time points.The residue is reconstituted with 1% DMSO complete culture solution and then added to HK-2、HMC、HRGEC for toxicity experiments.The MTT colorimetric method was used to determine the inhibitory effect of drug-containing serum on the proliferation of three kinds of cells,the results showed that compared with the blank serum(0 hour),the inhibitory effect of the drug-containing serum on cell proliferation increased significantly after 3 hours(p < 0.05).Chapter 3 Study on Serum Medicinal Chemistry of Tripterygium Wilfordii TabletsSerum drug chemistry method and high-performance liquid chromatography-mass spectrometry technology were used to analyze the blood components of drug-containing serum.94 peaks and 104 compounds were extracted,of which 82 peaks were derived from prototype drugs,12 were metabolite peaks,and 70 compounds were identified preliminarily.Chapter 4 Study on the spectrum toxicity relationship of drug-containing serum based on partial least squaresWith the partial least square regression analysis,the peak area of the fingerprint of the serum containing medicine at different time points was "X",and the cell proliferation inhibition rate was "Y".Establish the relationship between the content of the migrating components in the serum and the toxicity at each time point.Determination of toxic components by standard regression coefficient and VIP value given by software.The main toxic peak of HK-2 was B89、B79、B86、B21(tripterygiumine L)、B77、B72,HMC was B91(triptobenzene H)、B94、B93(triptobenzene A or triptoquinone D or triptobenzene M)and HRGEC was B80、B89、B88、B39.In this paper,the nephrotoxic substance basis of Tripterygium wilfordii polyglycoside tablets was clarified,which provided guidance for the establishment of scientific quality control methods of Tripterygium wilfordii preparations,the guidance of production process optimization,and the realization of synergism and toxicity reduction. |
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