The Effect Of Eucommia Ulmoides Water-extraction On Liver Damage Of Filial Mice Injured By Heroin

Objective:To investigate the influences of Eucommia ulmoides water-extraction on liver damage of filial mice injured by heroin.120 filial mice are intraperitoneal injected escalation doses of heroin at(1~5 d,6~10 d,11~15 d).Meanwhile,Eucommia ulmoides water-extraction intragastric administration was injected into filial mice to protect the liver.And some indices were measured,including filial mice weight,liver weight,structure of hepatic tissues,blood biochemical index,the activities of thioredoxin reductas(TrxR)and myeloperoxidase(MPO),the content of malonaldehyde(MDA),expression of Caspase-3 and Survivin protein.Methods:120 1-week-old mice were chosen and randomly divided into 4 groups: control,heroin,EUO 1 and EUO 2 groups.Each group has 30 mice.The heroin and EUO groups were intraperitoneal injected with the different doses heroin solution for 15 days,twice a day(8:00 am,16:00 pm).The 1~5 days,6~10 days,11~15 days were respectively injected with 2 g/L,3 g/L,4 g/L of heroin.And physiological saline was injected to the control group with same amount doses.After 1 hour of heroin injection,EUO 1 and EUO 2 were intragastrically administered with 0.6 g/mL and 1.8 g/mL Eucommia ulmoides water-extraction.The control group and the heroin group were intragastrically administered with same amount doses of physiological saline.Ten mice were sacrificed for each group of 5,10 and 15 d,and the changes of body weight and liver weight were measured and detected.Colorimetric method was used to detectγ-GT,AKP,ALT,AST activity in plasma of filial mice and changes from TrxR,MPO activity and MDA content in liver tissues of filial mice,the changes of liver tissue structure of filial mice were observed by optical microscope.SABC-immunohistochemical method was used to detect the changes expression of Caspase-3 and Survivin protein in liver tissues of filial mice.Results:1.Changes in body weight and liver weight of the filial mice: Compared with the control group,the body weight and liver weight of the heroin group were decreased in different degrees after injection of heroin.Eucommia ulmoides water-extraction wasintragastrically administered to the filial mice to protect,the EUO 1,2 groups of body weight and liver weight increased,however,these groups were lower than the control group.2.The changes of γ-GT,AKP,ALT and AST activities in the plasma: compared with the control group,the activities of γ-GT,AKP,ALT and AST of plasma in the heroin group were significantly increased with the increase of heroin injection time.The levels of γ-GT,AKP,ALT,and AST in plasma of the EUO 1,2 groups were significantly lower than those in the heroin group,but higher than those in the control group.3.The changes of TrxR,MPO activity and MDA content in the liver tissues of filial mice: compared with the control group,the TrxR activity in the liver tissues of the heroin group was significantly lower than that in the control group,and the activity decreased with the increase of the injection time of heroin.MPO activity increased significantly and increased with the increase of heroin time.However,the TrxR activity in the liver tissues of the EUO 1 and 2 groups was increased in different degrees compared with the heroin group.The MPO activity was reduced compared with the heroin group.The MDA content in the liver tissue of the heroin group was increased compared with the control group in each period,and the MDA content increased with the increase of heroin time,but in the liver tissue of MDA content of EUO 1 and 2 groups was reduced compared with the heroin group.4.Changes in liver tissue structure of the filial mice: The liver tissue of the control group was intact,the staining was uniform,the hepatic cell cord was regular,the hepatic sinus was normal,the hepatic lobule was intact,and the hepatocytes morphological structure was clearly visible.Compared with the control group,the liver tissue of the heroin group had different degrees of damage,the hepatic sinus was significantly bigger,some endothelial cells fell off,the hepatocytes showed obvious swelling,the nucleus was obviously pyknosis,and the hepatocytes were loose,vacuolation,degeneration,necrosis and other phenomena appear in the hepatocytes.The liver tissue damage of the EUO 1,2 groups was significantly reduced,the liver cells were arranged neatly,the hepatic sinus were significantly smaller,and the hepatocytes nuclear pyknosis and hepatocyte vacuolation were reduced.5.Changes in Caspase-3 and Survivin protein expression in the liver tissue of filial mice: The histochemical results of the SABC-immunized showed that the expression of Caspase-3 and Survivin in the liver tissue of the control group was extremely low,and the number of hepatocytes was extremely small.And the staining of control group was shallow;the expression of Caspase-3 and Survivin protein in the heroin group was significantly higher than the control group,and the number of staining cells in the liver increased with the increase of heroin injection time.The staining was dark brown;The EUO 1 and 2 groups whose expression levels of Caspase-3 and Survivin in the liver tissues were lower than those in the heroin group,and the number of stained cells in the liver of the filial mice was significantly reduced.Conclusions:1.Eucommia ulmoides water-extraction affected the body weight and liver weight of heroin liver injured filial mice.Heroin enters the body to stimulate the central nervous system of the filial mice through the blood-brain barrier and opioid receptors,resulting in euphoria,vertical tail running,increasing activity,increasing body material consumption.And heroin can directly damage the gastrointestinal tract of filial mice.The reduction of ingestion leads to the reduction of filial mice body weight.In addition,when the heroin is metabolized in the liver,it causes serious damage to the liver,affecting the normal metabolic activity of the liver of the filial mice,resulting in the decrease in body weight and liver weight.However,Eucommia ulmoides water-extraction can supplement the energy consumption of the filial mice and reduce the damage of heroin to the liver,so that the body weight and liver weight of the filial mice increase.2.Eucommia ulmoides water-extraction affect γ-GT,AKP,ALT and AST activities in the plasma of filial mice with heroin injured.In the process of heroin metabolism,the liver has accumulated a large amount of active oxygen free radicals.Active oxygen free radicals affect the activity of cell antioxidant enzymes,and result in liver cell lipid peroxidation damaging to liver blood circulation,liver cell membrane permeability increased significantly,the γ-GT,ALT,AST and AKP released into the blood,the γ-GT,AKP,ALT and AST activity of plasma was increased.However,Eucommia ulmoides water-extraction can effectively inhibit cell lipid peroxidation andmaintain cell membrane integrity,thereby reducing γ-GT,AKP,ALT and AST activities in the plasma.3.Eucommia ulmoides water-extraction affect the antioxidant enzyme activities in liver tissues of filial mice with heroin injured.After the intrauterine injection of heroin in the filial mice,heroin destroyed the antioxidant system and reduced its ability to scavenge free radicals.The activity of TrxR in the liver tissue of the filial mice was decreased,the activity of MPO was significantly increased,and the content of MDA was increased.Eucommia ulmoides water-extraction has strong ability to scavenge reactive oxygen free radicals,inhibit lipid peroxidation of hepatocytes,effectively reduce hepatocyte damage,thereby increasing TrxR activity,significantly lowering MPO activity,and lowering MDA content.4.Eucommia ulmoides water-extraction affect the liver tissue structure of filial mice with heroin injured.After injection of heroin,the heroin was dissolved into morphine and morphine accumulated in the liver,causing different degrees of damage to the liver tissue structure.The damage is more serious with time flies,while Eucommia ulmoides water-extraction can inhibit hepatocytes apoptosis and reduce liver cell damage.5.Eucommia ulmoides water-extraction affect the expression of Caspase-3and Survivin protein in liver tissues of filial mice with heroin injured.The expression of Caspase-3 and Survivin protein in the liver of heroin group was significantly higher than the control group,indicating that heroin can increase the apoptosis of liver cells in the filial mice.Eucommia ulmoides water-extraction inhibited the expression of Caspase-3 and Survivin proteins in liver tissue,This indicated that Eucommia ulmoides water-extraction could inhibit hepatocytes apoptosis through caspase-3 mediated apoptosis pathway,thereby protecting the liver tissue.