Triptolide(TP),an active component of Tripterygium wilfordii Hook.f.(TWHF),has been widely used for centuries as a traditional Chinese medicine.However,the clinical application of TP has been restricted due to multita...Triptolide(TP),an active component of Tripterygium wilfordii Hook.f.(TWHF),has been widely used for centuries as a traditional Chinese medicine.However,the clinical application of TP has been restricted due to multitarget toxicity,such as hepatotoxicity.In this study,28 days of oral TP administration(100,200,or 400μg·kg^(-1)·d^(-1))induced the occurrence of cholestasis in female Wistar rats,as evidenced by increased serum levels ofγ-glutamyl transpeptidase(γ-GGT),alkaline phosphatase(ALP)and hepatic total bile acids(TBAs).In addition,the heptocyte polarity associated with the strcture of tight junctions(TJs)was disrupted in both rats and sandwich-cultured primary hepatocytes.Immunoblotting revealed decreased expression of the TJ-associated proteins occludin,claudin^(-1),and zonula occludens protein(ZO^(-1)),and downregulated m RNA levels of these TJs was also detected by real-time PCR.An immunofluorescence analysis showed abnormal subcellular localization of occludin,claudin^(-1) and ZO^(-1),which was also confirmed by transmission electron microscopy.Moreover,the concentration of FITC-dextran,a marker of paracellular penetration,was found to increase rapidly in bile increased rapidly(within 6 minutes)after treatment with TP,which indicated the functional impairment of TJs.Taken together,these results suggest that the administration of TP for 28 consecutive days to rats could induce cholestatic injury in the liver,and the increased paracellular permeability might play an important role in these pathological changes.展开更多
基金supported by the National Natural Science Foundation of China(Nos.81803784,81320108029,and 81573690)the"Double First-Class"Projects of China Phar-macutical University(No.2018GY06)。
文摘Triptolide(TP),an active component of Tripterygium wilfordii Hook.f.(TWHF),has been widely used for centuries as a traditional Chinese medicine.However,the clinical application of TP has been restricted due to multitarget toxicity,such as hepatotoxicity.In this study,28 days of oral TP administration(100,200,or 400μg·kg^(-1)·d^(-1))induced the occurrence of cholestasis in female Wistar rats,as evidenced by increased serum levels ofγ-glutamyl transpeptidase(γ-GGT),alkaline phosphatase(ALP)and hepatic total bile acids(TBAs).In addition,the heptocyte polarity associated with the strcture of tight junctions(TJs)was disrupted in both rats and sandwich-cultured primary hepatocytes.Immunoblotting revealed decreased expression of the TJ-associated proteins occludin,claudin^(-1),and zonula occludens protein(ZO^(-1)),and downregulated m RNA levels of these TJs was also detected by real-time PCR.An immunofluorescence analysis showed abnormal subcellular localization of occludin,claudin^(-1) and ZO^(-1),which was also confirmed by transmission electron microscopy.Moreover,the concentration of FITC-dextran,a marker of paracellular penetration,was found to increase rapidly in bile increased rapidly(within 6 minutes)after treatment with TP,which indicated the functional impairment of TJs.Taken together,these results suggest that the administration of TP for 28 consecutive days to rats could induce cholestatic injury in the liver,and the increased paracellular permeability might play an important role in these pathological changes.
