Effects Of Aristolochic Acid Ⅰ And CD40 Signal On The Human Umbilical Vein Endothelial Cells

Backgrounds: Aristolochic acid is one of the phenanthrene compounds belonging to the species of Aristolochia of the Aristolochiaceae, which has the ability of anti-tumor, enhance immune function and increase white blood cell count and so on. So aristolochic acid has been generally used in the treatment of rheumatic arthritis, venomous snake bite and purulence of wound. Since the 90s, more and more aristolochic acid nephropathy(AAN) caused by drugs containing AA has been reported. AA with various doses at different time can cause various histologic changes of kidney. The data in vivo and in vitro indicated that AAN may refer to the injure of renal tubule endothelial cells(RTECs), proliferation and activity of myofibroblast in the renal interstitium, inflammatory cell infiltrate, transdifferentiation of RTECs and lead to renal tubule interstitial fibrosis. Recently, endothelial cells play an important role in the pathogenesis process of chronic kidney disease(CDK). But there are less reports about the effects of aristolochic acid on endothelial cells.Objective: To explore the effects of aristolochic acid I (AAI)on the human umbilical vein endothelial cells (HUVEC) by studying the proliferation, apoptosis, secretion and the transdifferentiation of HUVEC.Methods:1) The proliferation of HUVEC was researched by use of MTT method. 2) The apoptosis are detected with Flow cytometry (FCM). 3) The expression of TGF-β1 were measured by Elisa. 4) The endothelial marker Flk-1 and the smooth muscle actinα(α-SMA) on the HUVEC were studied by immuocytochemistry.Results: 1) After HUVEC were treated with AAI of 80μg/ml for 72 hours, the proliferation was suppressed obviously (P<0.05). 2) The apoptosis all increased at the level of 20μg/ml,40μg/ml,80μg/ml. 3) The aristolochic acid I at the level of 20μg/ml can promote the secretion of TGF-β1. 4) 5C11 at the level of 2.5μg/ml can increase the releasion of 2.5μg/ml significantly. 5) Neither down regulated Flk-1 nor up regulatedα-SMA were detected.Discussion: Recently, renal vascular endothelial cells injure play an important role in renal tubule interstitial fibrosis. AA can exert direct toxic effect on endothelial cells that suppressed proliferation and induced apoptosis with a lower level. The aristolochic acid I can promote the secretion of TGF-β1, which illustrated that TGF-β1 may involve in the injure of endothelial cells in AAN.Conclusion: Aristolochic acid I can supress the proliferation, cause the apoptosis of HUVEC and enhance the production of cytokine TGF-β1 related fibrosis which provide the new theory of microvascular injure in AAN.