Study On Correlation Between Toxicity And Effect Of Anthraquinone And Tannin Extracted From Rhubarb On HK-2 Cells

Objective The goal of this thesis is to explore correlation between toxicity and effect of anthraquinone and tannin extracted from Rhubarb, and the influencing factors of dose and different conditions of HK-2 cells are to be concerned about, what is more significant, this study may provide demonstration for researches on correlation between toxicity and effect of other toxic TCM.Methods The study on toxicity of anthraquinone and tannin extracted from Rhubarb, emodin and EGCG on HK-2 cells was carried out in different dose, and the change of cell morphology, OD of LDH, cell cycle and apoptosis were detected. Furthermore, HK-2 cells that were damaged at some level by exposing to H2O2 were pretreated with anthraquinone and tannin extracted from Rhubarb, emodin and EGCG,the effect of those drugs on impaired HK-2 cells were detected. By combinding with experimental data and traditional theory of TCM, the material basic, influencing factors and mechanism of correlation between toxicity and effct of Rhubarb were analyzed.Results (1) Anthraquinone extracted from Rhubarb and emodin possed some toxic on HK-2 cells, the toxic manifestation included:inhibition of cells proliferation, cell shrinkage and vacuolation, LDH leakage were increased, the cell cycle was blocked and induced apoptosis at different level. This result illustrated anthraquinone and emodin may be the materials that damage the kidney. The IC50 of anthraquinone extracted from Rhubarb is about 20.62mg·L-1 and the IC50 of emodin is about 120μmol·L-1. The main toxic mechanism of anthraquinone extracted from Rhubarb and emodin is to induce apoptosis and block the transformation of cell cycle from S period to G2/M period.(2) The toxicity of tannin extracted from Rhubarb on HK-2 cell was little, only leaded to lower grade change of cell shape, the inhibition ratio was 20% when the dose was 200mg·L-1.The percentage of anthraquinone and tannin extracted from Rhubarb is 1:2 approximately, contrast to the toxicity of anthraquinone, tannin is safe when the Rhubarb is administrated. The early apoptosis was induced by using EGCG when the dose exceed 80μmol·L-1.(3) In the condition of HK-2 cells were damaged by H2O2, anthraquinone extracted from Rhubarb protected the cells by improving the cell shape and activity, the OD was descended, especially, the apoptosis of cell was improved significantly. The effect of anthraquinone was concerned with the dose,15mg·L-1 anthraquinone mainly protected the cell, toxicity was detected when the dose was 30 mg·L-1, but the tolerance to drug toxicity of damaged cells was enhanced. Emodin represented the toxicity mainly by inducing advanced stage apoptosis and necrosis.(4) In the condition of HK-2 cell were damaged by H2O2, tannin extracted from Rhubarb protected the cells and the tolerance of damaged cells to drug toxicity was enhanced. The inhibitor ratio changed lighty and the OD of LDH was descended. EGCG protected the damaged cell by improving the early stage apoptosis and necrosis.Conclusions Base on the experiment on the toxicity and effct of Rhubarb, combind with the idea of correlation between toxicity and effect of toxic TCM, the thesis provide the strategy to the research on correlation between toxicity and effect of toxic TCM:The influencing factors of correlation between toxicity and effect are varied, but dose and different conditions play important roles. The correlation between toxicity and effect of anthraquinone and tannin extracted from Rhubarb include some aspects as follows:(1) Different materials may have different toxicity. Anthraquinone extracted from Rhubarb and emodin are more toxical than tannin extracted from Rhubarb and EGCG.. Anthraquinone extracted from Rhubarb and emodin change the cell shape, enhance the inhibition ratio and OD of LDH, block the cell cycle and induce the apoptosis and necrosis.(2) Different materials may have similar effect. Anthraquinone extracted from Rhubarb and emodin all protect the HK-2 cells damaged by H2O2. They all can maintain the normal cell shape, descend the inhibition ratio and cell apoptosis et al.(3) The same materials may have different toxicity and effect in different conditions. The tolerance of HK-2 cells to toxicity of anthraquinone and tannin extracted from Rhubarb are enhanced, anthraquinone and tannin extracted from Rhubarb mainly protect the damaged cells contrast to the normal condition at the same dose. The toxicity of emodin on damaged cells is more significant than the normal condition but EGCG protect the damaged cells.(4) The same materials may have different toxicity and effect in different dose. The toxicity of anthraquinone extracted from Rhubarb is little when the dose is lower than 30mg·L-1 and enhanced significantly when the dose is 60 mg·L-1, the inhibition ratio, the apoptosis and necrosis of HK-2 cells alter notablely.(5) The study may provide reference to explore correlation between toxicity and effect of other toxic TCM.