The Effect And The Mechanism Of Immunomodulating Peptide On Alcoholic Damage Cells Model

Objective: 1. To observe whether alcohol stimulation can induce normal liver cell(LO2) to produce fatty degeneration damaged mitochondria and endoplasmic reticulum stress(ESR) in vitro in human. 2. To research whether immunomodulating pepide(PGPIPN) can reduce liver cell damage and to explore the molecular mechanism.Methods: 1. Established stable model of alcoholic injury liver cells with screening concentration: MTT assay was used to screen the optima concentration of alcoholic and PGPIPN. Normal liver cells(LO2) were treated with alcoholic and different concentrations of PGPIPN for 3 months. 2. Oil Red O staining was used to detect the degree of LO2 cells steatosis. Transmission electron microscope was observed the mitochondria,nucleolus and other ultratructural. 3. The expressions of genes of fat synthesis related genes(ACC), damaged mitochondria related genes(Cyt C,Caspase-3) and endoplasmic reticulum stress related genes(GRP-78,GHOP) were tested by RT-PCR. 4. The expressions of protein of fat synthesis related genes(ACC), damaged mitochondria related genes(Cyt C,Caspase-3) and endoplasmic reticulum stress related genes(GRP-78,CHOP) were tested by Western Blotting.Result: 1. MTT screened for alcohol concentrations was 0.5%,the optimal drug concentrations for PGPIPN was 1×10-4g/L、1×10-3 g/L、1×10-2g/L, and then made the successful model. 2. Compared with normal liver cells, the alcohol control group of oil red O staining was steatosis, electron microscopy showed severe swelling of mitochondria; and PGPIPN groups can alleviate fatty degeneration, slow swelling of mitochondria. 3. In PGPIPN groups the expression of ACC, Cyt C, Caspase-3, GRP-78, CHOP at m RNA levels were decreased. 4. In PGPIPN groups the expression of ACC, Cyt C, Caspase-3, GRP-78, CHOP at protein levels were decreased.Conclusion: 1. PGPIPN can reduce alcoholic fatty degeneration and cell damage of the liver cells. 2. The mechanism maybe relate to slow fat synthesis, reduce mitochondria damage and relieve endoplasmic reticulum stress.