Correlation Between The Effect Of Neuritin Expression Level On Hsp60 In Hepatocytes

Objective: To explore the effect of Neuritin at different expression levels on HSP60 in hepatocytes and the molecular mechanism of regulation of hepatocyte apoptosis.Methods:Twenty plates of 100 mm petri dishes containing Buffalo Rat Liver(BRL)hepatocytes were randomly divided into 4 groups,namely the Neuritin low expression group,the Neuritin high expression group,the Hsp60 low expression group,and the Hsp60 high expression group.Each of the above large groups was divided into five groups: the control group(n = 1)and the experimental group(n = 4).No treatment in the control group.The experimental group in the Neuritin low expression group was given 0.1ug / ml and 0.3ug / ml respectively.ml,0.5ug / ml,0.7ug / ml of Neuritin antibody;in the experimental group of Neuritin high expression group,0.5ug / ml,1.0ug / ml,1.5ug / ml,2.0ug / ml of Neuritin protein were given respectively;Hsp60 low expression The experimental group in the group was given Hsp60 antibodies of 0.6ug / ml,0.8ug / ml,1.0ug / ml,and 1.2ug / ml respectively;the experimental group in the Hsp60 high expression group was given 10 × 10-3 ?g / ml,50 × 10-Hsp60 protein at 3 ?g / ml,100 × 10-3 ?g / ml,200 × 10-3 ?g / ml,and the expression change level of the target protein under the corresponding intervention conditions were measured by Western blotting after 48 hours.The location and co location of CPG15 and heat shock protein 60 were determined by immunofluorescence,and finally the possible molecular mechanism of the two in liver cells was discussed.Results:(1)Relative to blank group,Western blotting showed that the underexpression and overexpression of Neuritin protein in BRL cells in the experimental group gradually decreased and increased,while the content of Caspase-3 gradually increased and decreased.The difference between the two is statistically important(P <0.05);Hsp60 protein expression The levels were basically the same.The change of CPG15 was negatively correlated with the change of Caspase-3(r =-0.993,P <0.05).(2)Relative to blank group,Western blotting showed that the underexpression and overexpression of Hsp60 protein in BRL cells in the experimental group gradually decreased and increased,while the expression level of Caspase-3 protein was gradually increased and decreased.The difference between the two is statistically important(P <0.05);the expression level of Neuritin basically the same.The change of Heat shock protein was negatively correlated with the change of Caspase-3(r =-0.996,P <0.05).(3)Neuritin and Hsp60 are located on the cell membrane after Merge in cellular immunofluorescence.Conclusion:(1)Immunofluorescence test showed that Neuritin and Hsp60 were co located in the cell membrane,suggesting that there was interreaction between Neuritin and Hsp60.(2)Neritin and caspase-3 showed the opposite change trend,Hsp60 and caspase-3 also showed the opposite change trend,so it is speculated that neritin and Hsp60 may participate in the anti apoptosis regulation mechanism of hepatocytes through some synergistic effect.