A Study On The Effect Of The Ca²⁺ Signal Transduction In The Pathogenesis Of Acute Alcoholic Liver Injury

Purpose: According to the parenchymal liver cells isolation, purification and culture in vitro, analyzing the growth of primary hepatocyte, to study the Hepatocytes Toxicity of Rat and variation of intracellular dissociative Ca²⁺ induced by acetaldehyde, and the interventional effect of Ca²⁺ chelator, approach to the injury pathogenesis of liver cells induced by acetaldehyde and the protective effect of Ca²⁺ chelator.Method: To culture the rat primary hepatocytes for 24h and 48h in the culture medium including 300mmol/L acetaldehyde, and to observe these morphological changes by the phase contrast microscope. The rat primary hepatocytes were divided randomly into control group, treatment group with acetaldehyde, combined effection group with acetaldehyde and Fluo-3/AM. Laser Scanning Confocal Microscope was used to analyze the [Ca²⁺]i in the liver cells and the variation of Mitochondrial Transmembrane Potential (ΔΨm).Result: With the phase contrast microscope, the cells of control group have characters of easily refracting light and stereoscopic impression. The hepatocytes appear swelling, deep color, rough particles, partial cell detachment which were radial after 24-hour-treatment, and sparse cytoplasm, karyopycnosis, breaking cell membrane after 48-hour-treatment. By the treatment with acetaldehyde, with the concentration increasing,the [Ca²⁺]i increased, but contrary toΔΨm. Adding Fluo-3/AM with acetaldehyde, liver cells' mean fluorescence intensity andΔΨm increased more greatly than another groups.Conclusion: 1. Alcohol causes a seriously polarity disorder and forms different degrees of pathological status. Thus it investigates the pathogenesis of hepatopathy. 2. It may be related to the increase of the [Ca²⁺]i in hepatocytes that acetaldehyde induces the cytotoxicity. Ca²⁺ chelator can inprove and protect the [Ca²⁺]i homeostasis, and also release the cytotoxicity.