Objective To study the apoptosis-inducing effect of microcystin-LR (MC-LR) on primary cultured rat hepatocytes and the role of reactive oxygen species (ROS) in this effect.
Methods Primary cultured rat hepatocytes were obtained through collagenase perfusion method, resuspended in William's E medium, and cultured in orifice plates coated with rattail collagen at 37℃, 5% CO2. They were divided into 1 control group and 4 dose groups, and treated with gradient concentrations of MC-LR. Neutral red colorimetric assay was used to detect the viability of hepatocytes, and flow cytometry to detect apoptosis as well as ROS level in hepatocytes.
Results It was observed that the rat hepatocytes in two groups (MC-LR 1& #215;10-5, 1& #215;10-6 mol/L) almost totally died out, and in one group (MC-LR 1& #215;10-7 mol/L) died partly by neutral red colorimetric assay; and that MC-LR in duced rapid apoptosis and ROS formation by flow cytometry.
Conclusion MC-LR induces rapid massive ROS in primary cultured rat hepatocytes, which may be one of the mechanisms that MC-LR induces apoptosis and death of the cells finally.
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