| Salvia miltiorrhiza Bge.,a common traditional Chinese medicine,is widely used in the treatment of various diseases especially cardiovascular and cerebrovascular diseases and have good efficacy.It has a large market demand.However,in the market of Chinese medicinal materials,the source of Salvia miltiorrhiza is confused,the primary processing and production processing are different.There is no unified and standardized standard to guide from planting,initial processing to the production of finished products for Salvia miltiorrhiza Bge.,which results in the quality of Salvia miltiorrhiza is varying in the market,and even a large number of substandard Salvia miltiorrhiza circulating in the market.In this paper,the effects of Salvia miltiorrhiza Bge.Extracts with sweated or non sweated processed on P-gp in vitro and in vivo were studied in order to provide a scientific basis for the initial processing of Salvia miltiorrhiza.Objective: Firstly,To study the effects of Sweating and non sweating Salvia miltiorrhiza extracts on the function and expression of P-gp in HepG2 cells.Then,the sweating and non-sweating Salvia miltiorrhiza extracts were administered to rats to investigat their effects on the expression of P-gp in liver,intestine,brain,kidney and heart of rats.Methods: In vitro,MTT assay was used to determine the optimum concentration of total phenolic acid and tanshinone extracts from sweating and non-sweating Salvia miltiorrhiza for HepG2 cells.The effects of Salvia miltiorrhiza extracts on P-gp function of HepG2 cells were studied by Rho-123 assay.Then,RT-PCR and Western bloting were used to study the gene and protein expression of P-gp in both sweating and non-sweating Salvia miltiorrhiza extracts.In vivo,the effects of total phenolic acid extract and tanshinone extract of sweating and non-sweating Salvia miltiorrhiza on P-gp expression in liver,intestine,brain,kidney and heart of rats were studied by RT-PCR and Western bloting.Results: The concentration of sweating and non sweating total phenolic acid extract and tanshinone extracts of Salvia miltiorrhiza on HepG2 cells was determined by MTT assay.The concentration of total phenolic extracts was 20,100,500 μg/mL;the concentration of tanshinone extracts was 2,10,50 μg/mL.Rho-123 assay was used to detect the effects of total phenolic extracts and tanshinone extracts of sweating and non sweating Salvia miltiorrhiza on P-gp function in HepG2 cells.The results of accumulation and efflux experients showed that sweating and non-sweating total phenolic extracts of 20 and 100 μg/mL had no effect on P-gp in HepG2 cells,but had a certain induction effect at 50 μg/mL,and the effects of sweating was stronger than that of non-sweating.Tanshinone extracts of sweating and non sweating Salvia miltiorrhiza had no effects on P-gp of HepG2 cells at the concentration of 2 μg/mL.When the concentration was 10 and 50 μg/mL,it inhibited P-gp in HepG2 cells,and with the increase of concentration,the inhibitory effect is stronger.Under the same concentration,tanshinone extract has stronger inhibitory effect on sweating than non-sweating Salvia miltiorrhiza.The results of RT-PCR and Western bloting were basically consistent with the results of Rho-123 assay.Finally,RT-PCR and Western bloting were used to study the effects of sweating and non-sweating total phenolic extracts and tanshinone extracts on the expression of P-gp in liver,intestine,brain,kidney and heart of rats.The total phenolic acid extracts of Salvia miltiorrhiza had no effects on the expression of P-gp in rat liver,intestine,brain,kidney and heart.The expression of P-gp in liver,intestine,brain and kidney of rats was inhibited by sweating and non-sweating tanshinone extracts.and sweating is stronger than that of non-sweating Salvia miltiorrhiza extract.But it has no effect on the expression of P-gp in heart.Conclusion: The vitro and vivo experiments showed that the normal concentration of total phenolic extracts of sweating and non-sweating Salvia miltiorrhiza had no effect on P-gp function and expression.Increased concentration may induce P-gp function and expression,and the inducing effect of sweating is stronger than that of non-sweating total phenolic acid extracts.The low concentration of sweating and non-sweatingtanshinone extracts has no effects on the function and expression of P-gp.The normal concentration has an inhibitory effect.With the increase of concentration,the inhibitory effect is stronger.The Sweating tanshinone extracts is stronger than non-sweating in inhibiting P-gp.This experiment can potentially provide some scientific basis for the primary processing of Salvia miltiorrhiza.And may provide a theoretical basis for the combined use of Salvia miltiorrhiza and its extracts for multi-drug resistant drugs such as anti-cancer and anti-tumor. |
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