Study On The Effect Of Indoxyl Sulfate On Proliferation And Calcification In Vascular Smooth Muscle Cell

Objective:To investigate the effect of indoxyl sulfate(IS) on prol iferation and calcification in vascular smooth muscle cell (VSMC) and the relationship between oxidative stress and this effect. Methods:The effects of IS on calcification and proliferation in VSMC were studied respectively. Cells in each experiments were divided int8groups:normal group,100μMIS treatment group,300μMIS treatment group,500μM IS treatment group,10μM simvastatin+normal group,10μM simvastatin+100μM IS treatment group,10μM simvastatin+300P M IS treatment group and10μM simvastatin+500μM IS treatment gr oup.Calcium deposition in cells was measured by BCA.Osteocalcin level s were detected by radioimmuunityassay. The cell proliferation in VSMC was detected by WST-1method. Themalondialdehyde (MDA) content was de tected by thiobarbituric acid(TBA) method.Results:1. There was no significant difference in the calcium deposit ion between normal group and100μM IS treatment group. Compared with normal group, the calcium deposition in300μM IS treatment group and500μM IS treatment group were significantly increased(P<0.05and P<0.01, respectively).There was no significant difference in the calciu m deposition between100μM IS treatment group and10uM simvastatin+100μM IS treatment group. Compared with the same concentration IS g roup, the calcium deposition in10μM simvastatin+300μM IS treatme nt group and10μM simvastatin+500μM IS treatment group were signi ficantly decreased(P<0.05). Compared with normal group,the Osteocalci nlevels of supernatant in100μM IS treatment group,300μM IS treatme nt group and500μM IS treatment group were significantly increased(P <0.01). There was no significant difference in the Osteocalcinlevels of supernatant between100μM IS treatment group and10μM simvasta tin+100μM IS treatment group. Compared with the same concentration IS group, the Osteocalcinlevels of supernatant in10μM simvastatin+300μM IS treatment group and10μM simvastatin+500μM IS treatme nt group were significantly decreased(P<0.05). Compared with normal g roup, the MDA content in100μM IS treatment group,300μM IS treatment group and500μM IS treatment group were significantly increased (P<0.05,P<0.01and P<0.01, respectively). Compared with the same concentrat ion IS group, the MDA content in10μM simvastatin+100μM IS treatm ent group,10μM simvastatin+300μM IS treatment group and10μM sim vastatin+500μM IS treatment group were significantly decreased (P<0.05,P<0.01and P<0.01, respectively). There was significantly positive correlation between the calcium deposition and MDA content (r=0.59, p <0.001)2. There was no significant difference in WST-1content between normal group and100μM IS treatment group. Compared with normal grou p, WST-1content in300μM IS treatment group and500μM IS treatment group were significantly increased(P<0.05and P<0.01, respectively). There was no significant difference in WST-1content between100μMI S treatment group and10μM simvastatin+100μM IS treatment group. Compared with the same concentration IS group, WST-1content in10uM simvastatin+300μM IS treatment group and10μM simvastatin+500μ M IS treatment group were significantly decreased (P<0.05). Compared w ith normal group, MDA content in100μM IS treatment group,300μM IS treatment group and500μM IS treatment group were significantly inc reased (P<0.05, P<0.01and P<0.01, respectively). Compared with the same concentration IS group, the MDA content in10μM simvastatin+100μ M IS treatment group,10μM simvastatin+300μMIS treatment group an d10μM simvastatin+500μM IS treatment group were significantly de creased(P<0.05,P<0.01and P<0.01, respectively). There was significant ly positive correlation between the WST-1content and MDA content (r=0.59, P<0.001) Conclusion:IS may promote proliferation and calcification in rat VSMCwith concentration-dependent manner, which may be possibly related w ith the increased oxidative stress induced by IS. Simvastatin can inh ibit this effect.