The Research On The Effects Of HGF On Endoplasmic Reticulum Stress In Liver Tissue Of Mice With Nonalcoholic Steatohepatitis

Objective:To establish mice model of NASH,through the observation of the NASH mice model confirmed the presence of endoplasmic reticulum stress(ER stress)in the disease process;By injecting of recombinant human hepatocyte growth factor(rhHGF),we can explore the effects of HGF on ER stress of NASH liver tissue,to provide a new theoretical idea for the clinical application of HGF in NASH patients.Methods:A total of 18 male SPF grade BALB/c mice were selected and fed with standard diet AIN93M(10% kcal Fat)for about 1 week.The model was established after the first week.The mice were divided into normal control group(n = 6)and high fat group(n =12).The control group still use the AIN93M(10% kcal Fat)while the high fat diet group were fed with TP23400 purification(60% kcal Fat)for 16 weeks.At the end of the seventeenth week,the high fat group was divided into NASH model group(n = 6)and HGF group(n = 6).HGF group mice were injected with rhHGF through tail vein,normal control group and NASH group were injected intravenously with saline.All injection volume was 0.5g/(kg · d).The injection was continuous of 14 days.After 12 hours overnight fasting,the body weight were measured.The serum were separated from the angular vein blood by centrifugation,and were measured the lever of alanine aminotransferase(ALT),aspartate aminotransferase(AST),triglycerides(TG),interleukin-1?(IL-1?),interleukin-6(IL-6)and tumor necrosis factor-?(TNF-?).The mice were killed and the liver wet weight was measured,calculated the liver index andobserved the liver morphology.The middle part of the liver tissue were sampled for HE dye to observe the pathological changes under the microscope and evaluate score with NAFLD activity score(NAS);All protein of liver tissue was extracted using Keygen total protein extraction kit,measuring the ER stress interacting protein lever of glucose regulatory protein 78(GRP78)and C/EBP homologous protein(CHOP).Results:1.Create NASH mice model successfully;2.The body weight and liver index of NASH model group was obviously higher than normal control group(P ? 0.05);3.The levers of ALT,AST,IL-6,IL-1 ? and TNF-? in NASH group were significantly higher than those in normal control group(P ? 0.05),HGF group is significantly decreased than NASH group(P ? 0.05);4.The TG level in the model group was obviously higher than normal control group(P ? 0.05),HGF group was higher than that of NASH group,(P ? 0.05);5.Normal control group HE staining of liver tissue sections showed clear lobules,hepatocytes and hepatic sinusoids were neatly arranged like parallel radiation,without lipid droplets and inflammatory cells.In NASH model group,lobular structure disordered and liver cells arranged disorderly,with a large number of lipid droplets and inflammatory cells infiltrating;Compared with NASH group,pathological changes in HGF group was significantly alleviated;6.The level measured by ELISA of GRP78 in the NASH model group was higher than that in the normal control group(P ? 0.05),the level of CHOP was higher than that in the normal control group,but there was no significant difference.The levels of the two proteins in HGF group were lower than those in NASH group significantly(P? 0.05).Conclusion:NASH mice model was established and it was observed that ER stress was involved in liver injury during the development of NASH;HGF may protect the liver with nonalcoholic steatohepatitis by inhibiting the endoplasmic reticulum stress reaction.