Study On Microstructure And Ultrastructure Of Velvet Protective Effects On Acute Liver Injury In Mice

In recent years various liver diseases is one of the major diseases threating human health, so that prevention and treatment of various types of liver disease have attracted great attention from scholars at home and abroad.The velvet is the traditional tonic products with abundant chemical composition and active composition,which has extensive phamraeology activity and medical treatment and health care function.In order to observe protective effects of velvet on acute liver injury in mice and to explore its possible mechanisms,in the study we dealt the mice with CC14,APAP,Alcohol and BCG+LPS to establish the models of chemistry,drug, alcohol,and immune acute liver injury.Mice(20g±1g) were randomly divided into blank group,liver-injury model group,EAV(extract by alcohol from velvet,2.0g/kg) group and EAV(1.0g/kg) group,velvet powder group and bifendate group,and prepared paraffin section and ultrathin section of electron microscopy,observed pathological changes of liver tissue and liver cell ultrastructure.Our study could provid the theoretical basis for its development of new drugs of anti-liver injury.1.The study on microstructure and ultrastructure of velvet protective effects on chemical liver injury in mice:(1) Velvet could lower the mice's liver coefficient obviously and reduce acute liver injury induced by CCl₄.Compared with the model group(6.16±0.15%) the effects of EAV groups(2.0g/kg,5.25±0.09%,1.0g/kg,5.21±0.12%) were the best(P＜0.01),and other treatment groups also had better effects.(2) HE section showed that in model group there were obvious steatosis,focal necrosis with inflammatory collections,hepatielobule's boundaries was unclear,celt swolled, the normal structure of cell disappeared,whereas the drug-treated groups relieved the necrosis,swelling,steatosis,inflammatory of hepatocytes.The effects of EAV(2.0g/kg) was better than other treatment groups,and there were no obvious necrosis and steatosis in the group.Statistical analysis showed that compared with blank group the CC14 group had significant difference(P＜0.01) and the mice acute hepatic injury experimential model was built very successfully.Compared with the model group EAV(2.0g/kg) and the positive control bifendate reduced the pathological changes significantly(p＜0.01),although in EAV(1.0g/kg) and velvet powder groups the pathological changes had been reduced,the difference is not significant(p＞0.05).(3) Ultrathin section of electron microscopy showed that in model group liver cell membrane damaged,mitochondria swollen and cristae disappeared or fractured,rough endoplasmic reticulum degranuled,chromatin condensated,fat droplets grown in number, microvilli of bile duct lost and glycogen reduced.Velvet could improve the condensation of chromatin,the expansion of mitochondria and RER,and other pathological changes,and improved the protein,fat and glycogen metabolism in the liver.The effects of EAV (2.0g/kg) was better than other groups,and had been near normal,the pathological changes in velvet powder and EAV low-dose groups also were reduced.(4) Velvet could reduce the lipid droplets and improve the liver fat metabolism.The effects of EAV(2.0g/kg) was better than other groups,near-normal liver architecture possessing higher hepatoprotective action,the effects of velvet powder was bad,the same effects occurred between BIF and EAV(1.0g/kg).(5) Velvet could deincrease damage of the bile duct and microvilli,and restore the function of secretion and excretion of liver,every treatment group worked well.2.The study on microstructure and ultrastructure of velvet protective effects on drug-induced liver injury in mice:() Velvet could lower the mice's liver coefficient and reduce acute liver injury induced by APAP.Compared with the model group(5.91±0.15%) mice's liver coefficient of EAV(2.0g/kg,5.42±0.09%) lowered significantly(P＜0.05),although in other treatment groups mice's liver coefficient had been reduced,the difference was not significant (p＞0.05)(2) HE section showed that in model group liver cell cord disordered,there were patches necrosis of liver cell with inflammatory collections around central vein and area of focal necrosis,hepatielobule's structure was unclear,cell swollen,while the drug-treated groups relieved the necrosis,swelling,inflammatory of hepatocytes.Statistical analysis based on the degree of liver injury showed that compared with the model group EAV (2.0g/kg,1.0g/kg) and the positive control bifendate reduced the pathological changes significantly(p＜0.01),the effects of velvet powder worked worst(p＜0.05).(3) Ultrathin section of electron microscopy showed that in model group there was vacuole necrosis in hepatocytes,RER and mitochondria swollen,mitochondria's cristae reduced and electron density lowerd,nuclear membrane shrunk,nuclear matrix degenerated at the early stage of karyopyknosis.Capillary bile duct closed up,small amount of lipid droplets,neutrophils and fibroblasts can be seen in liver cells.In treatment groups pathological changes of hepatocytes were improved significantly,generally for this respect,the effects of EAV were better than velvet powder,EAV(2.0g/kg) worked as well as BIF.(4) Velvet could repair damage of organelle such as mitochondria,endoplasmic reticulum and inhibit the reduction of protein synthesis and disorder of lipid metabolism caused by liver injury,from our study we could draw that the effects of EAV(2.0g/kg) were better than other treatment groups,and organelle was no obvious changes and near blank group,but the velvet powder was the worst.(5) Velvet could increase the function of liver's secretion and excretion,every treatment group had a better role,and the mice treated with velvet agents had the lighter injury.3.The study on rnicrostructure and ultrastructure of velvet protective effects on the alcohol-induced liver injury:(1) Velvet could decreased the mice's liver coefficient obviously and reduce acute liver injury induced by Alcohol.Compared with model group(5.88±0.09%) EAV group (2.0g/kg,5.08±0.06%),velvet powder(5.37±0.09%) and BIF(5.18±0.07%) had significant differences(P＜0.01),and EAV(1.0g/kg) was worse(P＜0.05),.(2) HE section showed that in model group hepatielobule's boundaries was unclear, cell swollen and the worst displayed bolloning generation,there were scattered punctate necrosis around central vein,fatty degeneration of hepatocytes and inflammatory around area of necrosis or in sinusoid;while the drug-treated groups relieved the necrosis,swelling, steatosis,inflammatory of hepatocytes.In term of it the effects of BIF was slightly superior to EAV(2.0g/kg),and EAV(1.0g/kg) and velvet powder were a little worse.Statistical analysis showed that compared with the model group the effects of EAV(2.0g/kg) and BIF were better(p＜0.01),EAV(1.0g/kg) was a little worse(p＜0.05),and velvet powder was the worst(p＞0.05) but liver injury was relieved in every treatment group.(3) Ultrathin section of electron microscopy showed that in model group hepatic cells loose,organelle scattered,mitochondrial seriously swollen,membrane fusion,cristae reduced and giant mitochondria generated,rough endoplasmic reticulum did not change significantly with only slight expansion and degranulation.Nuclear membrane shrunk,chromatin marginated,there were a lot of fat droplets in liver cell and inflammatory,capillary bile duct closed up,there were collagen fibers hyperplasia in dieldrin gap.Treatment groups had relieved the above-mentioned pathological changes to different extents.For protecting liver's injury,the effects of EAV(1.0g/kg) were better than the velvet powder but worse than the EAV (2.0g/kg) and BIF(worked as well as EAV(2.0g/kg)).In addition velvet promoted hyperplasia of mitochondria,to speed up the metabolism of ethanol.(4) Velvet could inhibit fatty degeneration and ruduced the number of fat droplets,thus improved liver function of lipid metabolism.The effects of EAV(2.0g/kg) and BIF were the best,while EAV(1.0g/kg) and velvet powder were a little worse.(5) Velvet could alleviate cholestasis of liver in every treatment group obviously,the effects of velvet powder was the less worse.EAV and BIF could be seen improve effectively the the damage of bile duct and microvilli to restore the secretion and excretion function of liver.4.The study on microstructure and ultrastructure of velvet protective effects on the immunological liver injury:(1) Velvet could decreased the mice's liver coefficient obviously and reduce acute liver injury induced by BCG+LPS.Compared with model group(6.09±0.12%),EAV(2.0g/kg, 5.59±0.08%) and BIF(5.60±0.11%) had significant differences(P＜0.01),EAV (1.0g/kg,5.77±0.09%) and velvet powder(5.84±0.09%) was worse(P＞0.05).(2) HE section showed that in model group liver cell cord disorders,there were the existence of a large number of punctate necrosis with inflammatory and hemorrhage,nuclear size were different and showed shrinkage of different degrees.Eosinophilic enhanced,and there were mild fatty degeneration,there was siltation of blood in sinusoid,while the drugtreated groups relieved the necrosis,swlling,steatosis,inflammatory of hepatocytes.The results of statistical analysis showed that compared with the model group the effects of BIF and EAV(2.0g/kg) were significant different(p＜0.01),velvet powder(p＜0.05),because of dose-effect relationship in EAV(1.0g/kg) the difference was not significant(p＞0.05), meanwhile there was no difference between EAV(2.0g/kg) and BIF.(3) Ultrathin section of electron microscopy showed that in model group the number of small lipid droplets,glycogen,neutrophils and lymphocytes increased,microvilli of hepatocyte swollen,sinusoidal narrowed and sinus endothelial structure was unclear.The membrane structure of cell and organelle fused,mitochondrial swollen,the number of endoplasmic reticulum reduced,some endoplasmic reticulum had mild degranulation and swelling.Treatment groups had different relief of the above-mentioned pathological changes. The effects of EAV(2.0g/kg) and BIF surpassed that of EAV(1.0g/kg) and velvet powder,and EAV(2.0g/kg) group had been near blank group.(4) In terms of improving lipid metabolism velvet could inhibit fatty degeneration,and reduce accumulation of TG in the liver.The effects of EAV(2.0g/kg) was better than other treatment group and there were few fat droplets.(5) Velvet could increase the secretion and excretion function of liver,and alleviate cholestasis of liver,the every treatment group had a better role,and the mice treated with velvet agents had the lightst injury.EAV(2.0g/kg) were the best and close to the blank group, followed by the BIF group,the effects of EAV(1.0g/kg) and velvet powder was also evident,but less than the former two groups.