| Rhizoma Coptis and its main ingredient Berberine have many functions in antibacterial, anticancer, Antioxidant and so on. They have been widely used in clinics.However, Rhizoma Coptis and Berberine also showed some side effects in the treatment of diseases. Therefore, some scholars has been researched the basic toxicity about Rhizoma Coptis in China, but the toxic effects on cells of Rhizoma Coptis and Berberine have not been reported. In the experment, BRL cells, NRL cells and L929 cells cultured in vitro were used to investigate the toxicity and its mechanism, to evaluate risks of Rhizoma Coptis and Berberine at the cellular level, which would lay a foundation for future clinic applications of Rhizoma Coptis and Berberine.Rhizoma Coptidis and Berberine were diluted into 10 different concentrations, then co-culture with monolayers of BRL cells, NRL cells and L929 cells respectively. CCK-8 was used to detect the influence of Rhizoma Coptidis and Berberine on proliferation (or inhibition) of treated cells. On the basis of CCK-8 results, we designated the high inhibition rate of drug concentration (the inhibition rate of the three kinds of cells was about 75%, hereinafter referred to as the high dose), lower inhibition rate of drug concentration (the inhibition rate of the three kinds of cells was about 25%, hereinafter referred to as middle dose) and does not produce inhibition of drug concentration (the inhibition rate of the three kinds of cells was about 0%, hereinafter referred to as the low dose) respectively to three kinds of cells, and Annexin V-FITC and PI double dye were employed to detect apoptosis rate, JC-1 tested cell mitochondrial membrane potential changes. Results are as follows: 1. CCK 8 test results displayed that the concentrations of Rhizoma Coptis which was higher than 0.19mg/mL,1.56mg/mL,1.56mg/mL could inhibit cells proliferation on BRL cells, NRK cells, L929 cells, the concentrations of Berberine which was higher than 0.0338mg/mL,0.0338mg/mL,0.0169mg/mL could inhibit cells proliferation on the three of cells, the concentrations of Rhizoma Coptis which was lower than 0.0169mg/mL,0.0169mg/mL,0.0084mg/mL could promote cells appreciation on BRL cells, NRK cells, L929 cells, the concentrations of Berberine which was lower than 0.0169mg/mL,0.0169mg/mL,0.0084mg/mL could promote cells appreciation on the three of cells. The IC50 of Rhizoma Coptis and Berberine is NRK cells> L929 cells> BRL cells, the toxicity classification of Rhizoma Coptis is BRL cells> L929 cells> NRK cells, the toxicity classification of Berberine is BRL cells> L929 cells and NRK cells.2. Flow cytometry detection of apoptosis showed that the total rate of apoptosis (%) of BRL cells were 3.115±0.019,1.659±0.017 and 0.055±0.001, the total rate of apoptosis of NRK cells were 5.040±0.030,3.458±0.025 and 1.972±0.015, the total rate of apoptosis of L929 cells were 5.980±0.055,4.790±0.040 and 3.300±0.079, which were induced by high, medium and low concentrations of Rhizoma Coptis. The total rate of apoptosis of BRL cells were 3.493±0.006,1.923±0.013 and 0.864±0.009, the total rate of apoptosis of NRK cells were 4.906±0.030,2.344±0.015 and 1.958±0.015, the total rate of apoptosis of L929 cells were 5.554±0.021,4.421±0.009 and 3.784±0.045, which were induced by high, medium and low concentrations of Berberine. It showed Rhizoma Coptis and Berberine can induce apoptosis of BRL cells, NRK cells and L929 cells, with the drug concentration increases, the cell apoptosis rate increased, it showed a dose-dependent manner.3. The results of cells changes by JC-1 test mitochondrial membrane potential showd that the rate of red and green fluorescence of BRL cells were 0.336±0.036,2.479±0.025 and 6.114±0.053, the rate of red and green fluorescence of NRK cells were 0.669±0.031, 1.586±0.044 and 4.484±0.0312, the rate of red and green fluorescence of L929 cells were 1.070±0.064,1.260±0.031 and 1.700±0.034, which were induced by high, medium and low concentrations of Rhizoma Coptis. The rate of red and green fluorescence of BRL cells were O.388±O23,2.624±0.009 and 9.281±0.055, the rate of red and green fluorescence of of NRK cells were 0.540±0.036,0.911±0.028 and 3.170±0.042, the rate of red and green fluorescence of L929 cells were 0.801±0.042, 1.088±0.037 and 1.420±0.042, which were induced by high, medium and low concentrations of Berberine. It showed that Rhizoma Coptis and Berberine can change mitochondrial membrane potential of BRL cells, NRK cells and L929 cells.Conclusion:Rhizoma coptis and Berberine has certain toxic effects on NRK cells, BRL cells and L929 cells, the poisonous to the BRL cells was the largest, to the NRK cells of NRK was the minimum, to the L929 cell was medium, and high dose of Rhizoma coptis and Berberine can cause cell early apoptosis and mitochondrial membrane potential changes. |
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