| Aristolochic Acid Nephropathy (AAN), which is caused by intake of traditional Chinese medicines (TCM) containing aristolochic acids (AA), has gained wide concern throughout the world. However, a method to systemically and properly estimate the toxicity and the risk of TCM containing AA in a cytogenetic and molecular scale needs establishing in our country. Therefore, it is aimed to launch the establishment of such a method in this research.For this purpose, firstly, High Performance Liquid Chromatography (HPLC) was applied to quantify the content of AA in the extracts of 8 prescripts of TCM. And among them, single recipe Radix aristolochiae and compound recipe Longhui Pill were chosen to test their cell toxicity and genotoxicity respectively, by L5178Y/TK+/- Mouse Lymphoma Assay (MLA), due to their highest AA content.The result of quantification of AA content in selected TCM is that their AA content was between 0～4.72 mg/g. Among single recipe TCMs, the highest content of AA presented in Radix aristolochiae with 3.11512 mg/g, and among compound recipes, in Longhui Pill with 4.72044 mg/g. The result also shows that the HPLC method described has not only been simple, reliable, and accurate, but also met the needs of recovery rate and reoccurrence well.In the result of MLA, in the condition that AA contents in the solution of Radix arisotolochiae extract were 0, 25, 50, 100, 200μg/mL, the mutation frequency (MF) data of the tk gene of L5718Y3.7.2c-tk+/- mouse lymphoma cells were: 115.3×10-6, 303.1×10-6, 632.4×10-6, 1346.2×10-6, 2972.3×10-6 respectively. In the case of Longhui Pill, under similar condition, were 115.3×10-6, 122.9×10-6, 237.2×10-6, 586.7×10-6, 1150.8×10-6 respectively. Simultaneously, parameters of cell viability, namely relative survival (RS), relative suspension growth (RSG) and relative total growth (RTG) were in inverse relation with the concentration of extract of Radix Aristolochiae and Longhui Pill. The two facts indicate that both medicines bore cell toxicity and genotoxicity, which worsened as the concentration of extract rose. However, according to the data, in the situation of compound recipe Longhui Pill, compared with single recipe Radix aristolochiae, the stress of cell toxicity and genotoxicity was alleviated, which suggests that compound recipes may have detoxication effect towards AA. This phenomenon should be a valuable topic for further study in the future.In summary, a preliminary method, utilizing HPLC and MLA, has been established to estimate the toxicity of AA-containing TCMs in this research, and a step forward has been made in the study of the possibility for detoxication of AA by compound recipe TCMs. |
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