Study On The Hepatotoxicity Of Polygonum Cuspidatum And Reyanning Heji

Objective:The mice were given the maximum drugs concentration that the mice can tolerate,which were Polygonum cuspidatum raw product,water decoction,vinegar processed Polygonum cuspidatum,wine processed Polygonum cuspidatum,salt processed Polygonum cuspidatum,polydatin suspension,Polygonumcuspidatumanthraquinoneaqueoussolution,ReyanningHeji,ReyanningHeji?notcontainingPolygonum cuspidatum?.The effects of the drugs on the mice was observed after administration.Using of RTCA technology and HCA technology to detect AttenuatedeffectofPolygonumcuspidatumafterprocessingand compatibility;Using the RTCA technology and HCA technology to detect the toxicity of polydatin and Polygonum cuspidatum anthraquinone;Using the inverted microscope to observe cell morphology after Polygonum cuspidatum raw product,water decoction,vinegar processed Polygonum cuspidatum,wineprocessedPolygonumcuspidatum,saltprocessed Polygonumcuspidatum,polydatinsuspension,Polygonumcuspidatum anthraquinone aqueous solution,Reyanning Heji,Reyanning Heji?not containing Polygonum cuspidatum?acting on human normal liver cell L-02.Methods:acutetoxicitytest:Polygonumcuspidatumwater decoction,vinegar processed Polygonum cuspidatum water decoction,wine processedPolygonumcuspidatumwaterdecoction,saltprocessed Polygonum cuspidatum water decoction,polydatin suspension,Polygonum cuspidatum anthraquinone aqueous solution,Reyanning Heji and Reyanning Heji?Not containing Polygonum cuspidatum?.The mice was given the maximum drugs concentration it can tolerate.the administration volume is0.4ml/10g for three times in a day.The diet and excretion of the mice and the changes of Blood,lung,stomach,small intestine,liver,kidney and spleen were observed.Pathological tissues were observed in the altered organs.RTCA test:The cell concentration was 6×10⁴/ml by diluting.100?l cell suspension was added to each hole of E-16plate.The corresponding drug concentration was added to each hole after 24 hours.DMEM culture medium was added to the control hole and 1.25mg/ml acetaminophen solution was added to the positive hole.Then the E-16plate was put into the cell incubator for 56 hours.Morphological observation:The cell concentration was 6×10⁴/ml by diluting.100ul cell suspension was added to each hole of 96-well plate.The corresponding drug concentration was added to each hole after 24 hours.DMEM culture medium was added to the control hole and 1.25mg/ml acetaminophen solution was added to positive hole.Then the 96-well plate was put into the cell incubator for 24 hours.Each hole were gently washed with sterile PBS solution after 24 hours,and the morphological changes were observed under the inverted microscope.HCA evaluated Polygonum cuspidatum and Reyanning Heji on human normal liver cells L-02 toxicity test:10%FBS DMEM culture medium diluted cell suspension density to 6×10⁴/ml,96-well plate in each hole by adding 100ul cell suspension,about 24 hours after each hole by adding the appropriate concentration of drugs,the control hole by adding 10%FBS DMEMculturemedium,positiveholeaddedtothe1.25mg/ml acetaminophen solution,put three gas incubator continuous culture for 24hours,abandoned Liquid and medium,add the probe incubation 30min,on the machine detection.Results:Acute toxicity:The toxicity of drugs is low,which were Polygonum cuspidatum water decoction,vinegar processed Polygonum cuspidatum water decoction,salt processed Polygonum cuspidatum water products,wineprocessedPolygonumcuspidatumwaterdecoction,Reyanning Heji,Reyanning Heji?not containing Polygonum cuspidatum?,polydatinsuspension,Polygonumcuspidatumanthraquinoneaqueous solution.The MTD that mice can tolerate were 799.2g/kg,944.4g/kg,968.4g/kg,912g/kg,817.2g/kg,558g/kg,75.6g/kg,96g/kg.The mice of administration groups did not die.there was no significant difference between the administration groups and the blank group.RTCA experiment:The toxicity of Reyanning Heji is lower than Polygonum cuspidatum raw products and The toxicity Reyanning Heji?not containing Polygonum cuspidatum?is lower than Reyanning Heji.Acting on human normal liver cells L-02 for 24 hours,IC₅₀0 were 13.08mg/ml,11.81mg/ml,13.32mg/ml.Acting on human normal liver cells L-02 for 48hours,IC₅₀0 were 12.44mg/ml,11.97mg/ml,13.14 mg/ml.the toxicity of vinegarprocessed,wineprocessedandsaltprocessedPolygonum cuspidatum was reduced.Acting on human normal liver cells L-02 for 24hours,IC₅₀0 were 12.15mg/ml,15.23mg/ml,31.26mg/ml;Acting on human normal liver cells L-02 for 48 hours,IC₅₀0 were 12.00mg/ml,16.10mg/ml,69.97mg/ml.Toxicity can be obtained by comparative analysis:raw products>vinegar products>wine products>salt products.Morphologicalobservation:Undertheinvertedmicroscope,the positive hole significantly inhibited the proliferation of cells compared with thecontrolhole.Thecontrolholes?Polygonumcuspidatum,vinegar processed Polygonum cuspidatum,wine processed Polygonum cuspidatum,salt processed Polygonum cuspidatum,polydatin suspension,Polygonum cuspidatum anthraquinone aqueous solution,Reyanning Heji?not containing Polygonum cuspidatum?,Reyan ning Heji?showed a decrease in the number of cells with the increase of drug concentration and cells morphology changed significantly.HCA evaluateds Polygonum cuspidatum and Reyanning Heji on human normal liver cells L-02 toxicity test:10%FBS DMEM culture medium diluted cell suspension density to 6×10⁴/ml,96-well plate in each hole by adding 100ul Cell suspension,about 24 hours after each hole by adding the appropriate concentration of drugs,the control hole by adding 10%FBS DMEMculturemedium,positiveholeaddedtothe1.25mg/ml acetaminophen solution,put three gas incubator continuous culture for 24hours,abandoned Liquid and medium,add the probe incubation 30min,on the machine detection.HCA evaluates Polygonum cuspidatum and Ryanning Heji on human normal liver cells L-02 toxicity test:Polygonum cuspidatum,Polygonum cuspidatum vinegar products,Polygonum cuspidatum,Tyuriaceae Sunflower tablets,polydatinsuspension,Polygonumcuspidatum,Ryanning Heji,Ryanning Heji?not containing Polygonum cuspidatum?acts on normal human liver cells 24h,compared with the negative hole,the drug effect,the number of cells decreased mitochondrial membrane potential,cell area becomes smaller,and has a gradient Dependency.Conclusion:Polygonum cuspidatum through processing and Reyanning Heji had no obvious acute toxicity.The toxicity of Polygonum cuspidatum was reduced after processing and compatibility.