A Preliminary Study On The Toxicology Of Cantharidin Poisoning

Objective1.To explore the possible pathological changes of acute poisoning induced by cantharidin in mice.2.To investigate the possible mechanism of liver injury induced by cantharidin in mice and human hepatocytes.Methods1.The Kunming mice were given different lethal doses of cantharidin.Morphological changes of heart,lung,liver,spleen,kidney and brain were observed under microscope.2.The Kunming mice were orally given different doses of cantharidin for 14 days.The hepatic function and liver index was detected.Morphological changes were observed.Western blot was used to identify the expression of ATF-6,XBP1,GRP78,ATF-4,CHOP,BAX,Bcl-2,Caspase3,Caspase8 and Caspase9.Human LO2 hepatocytes were incubated with different concentrations of cantharidin.CCK-8 assay was used to detect the activity.Real-time PCR was used to quantify the mRNA expression of GRP78 and CHOP.Western blot was used to identify the expression of GRP78,ATF-4,CHOP,BAX,Bcl-2,Caspase3,Caspase8 and Caspase9.Results1.Cantharidin acute poisoning of parenchymal cell in mice induces degeneration and necrosis,vasodilation congestion,bleeding.2.Cantharidin up-regulated the levels of transaminases and liver index.Necrosis and apoptosis of hepatocytes in mice were aggravated in cantharidin-intervented groups compared to the control group,with a positive correlation of the does.The expression of ATF-6,XBP1,GRP78,ATF-4,CHOP,BAX,Caspase3,Caspase8 and Caspase9 was up-regulated and the expression of Bcl-2 protein was down-regulated(P<0.05).The proliferation of LO2 hepatocytes was inhibited in cantharidin-intervented groups.The expression of GRP78 and CHOP mRNA was increased in cantharidin-intervented groups.The expression of GRP78,ATF-4,CHOP,BAX,Caspase3,Caspase8 and Caspase9 was up-regulated and the expression of Bcl-2 protein was down-regulated in cantharidin-intervented groups(P<0.05).Conclusions1.Cantharidin can cause pathological changes in the heart,lung,liver,spleen,kidney,brain tissue damage,especially liver.2.Cantharidin can induce liver cell apoptosis by endoplasmic reticulum stress and damage hepatocytes.