Studies On Toxicodynamics And Toxic Mechanism Of Fructus Xanthii

Objective:To systematically dexcribe the spectrum of toxic effects, dose-response curve; and dose-time-respose relationship of Fructus Xanthii, and to expore the toxic mode of action (related)/toxic mechanism (cause) of Fructus Xanthii.Methods:To respectively prepare the water extract of stired Fructus Xanthii(sampleⅠ),70% alcohol extract of stired Fructus Xanthii(sampleⅡ), the water extract of crude Fructus Xanthii(sampleⅢ)and 70% alcohol extract of crude Fructus Xanthii(sampleⅣ). Using the method of the acute toxic test, to observe the four samples'acute toxicity of Fructus Xanthii in mice by one-time intragastric administration, reach the half lethal dose in mice (LD50) and its 95% confident limit with probability. To observe the toxic reaction of Fructus Xanthii stired water extract by the maximum dose of determination test on rat. To observe the specific or general toxic effects, death and mortality, and to identify the target organ toxicity and the organ's pathological changes. To observe the 3 months course of long-term toxicity of Fructus Xanthii stired water extract and the general behavior with the method of long-term toxic. To detect animal hematology, blood biochemistry and organ coefficient, and find target organ toxicity and the organ's pathological changes. To study on Fructus Xanthii stired water extract in the liver metabolism and toxicity mechanism using the primary hepatocytes.Results:①Acute toxicity test in mice The LD0 of sampleⅠ, sampleⅡsampleⅢand sample IV on mice by single oral administration were respectively 124,234,106,172g/kg; LD100 were 230,394,240,420g/kg; LD5o were 155.9,317.8,167.7,275.4g/kg; its 95% confident limit with probability were respectively 140.7～172.8,287.1～351.8,151.5～185.6,240.1～315.9g/kg. The signs of toxicity were as follow:still prone, abdominal breathing, vertical hair, tail and feet cyanosis, tremor, jumping, severe intermittent seizures, abdominal ground overly swing forward, hind legs twitch etc. Autopsy death mice, the main organs had unobviously pathological changes by both eyes observation. After HE staining and found that Fructus Xanthii stired water extract have hemorrhagic necrosis of liver spots, massive bleeding piece of liver congestion. Fructus Xanthii stired 70% alcohol extract have the lung, liver, gastric mucosa and heart congestive. Fructus Xanthii crude water extract have liver congestion associated with point spotty hemorrhage and loose cytoplasm, and Kinney, brain, heart, liver and gastric mucosa congestive. Fructus Xanthii crude 70% alcohol extract have droplet model of liver steatosis. The maximum dose of determination test of sample I on SD rat was 168.80 g/kg which was equal to 1012.8 times the clinical dose.②Long-term toxicity test After continuous administration of sample I for 1 month,3 months,15-day recovery, we detected hematology, blood biochemical parameters, organ's weight and organ coefficient weight determination. The results showed that blood biochemistry in rats have some influence on rat liver coefficient recoverable effects, pathological changes of organs has no effect.③Toxicity based on primary hepatocytes In the toxicity experiments of sample I and atractyloside based on the primary hepatocytes model, comparing with the control group, ALT,AST,TP,ALB,BUN,LDH of 2h,24h and 48h have the significant differences.ALT, AST and LDH of the samples are higher than the control group's, while BUN is lower.Conclusion:①The LD50 of sampleⅠ, sampleⅡ, sampleⅢand sample IV were respectively 155.9,317.8,167.7,275.4 g/kg in mice by one-time intragastric administration, which equal to 935.4,1906.8,1006.2,1653times the maximum clinical usage. The maximum dose of determination test of sampleⅠon SD rat was 168.80 g/kg.②SampleⅠlong-term toxicity test of 3-month, combined with weight and growth, hematology, blood chemistry, organ index, comprehensive analysis of pathology, suggesting that blood biochemistry in rats have some influence on rat liver coefficient recoverable effects, pathological changes of organs has no effect..③In the toxicity experiments of sampleⅠand atractyloside based on the primary hepatocytes model, comparing with the control group, ALT,AST,TP,ALB,BUN,LDH of Oh,2h,24h and 48h have the significant differences. ALT, AST and LDH of the samples are higher than the control group's, while BUN is lower. It shows that Fructus Xanthii do have some liver damage by the consentration set in this study.