The Protection Of Angelica Sinensis Polysaccharide On Liver Histological Structure In Mice Injured By Ephedrine

Objective:To explore the protection of Anglica Sinensis polysaccharide against mice hepatic tissues injured by ephedrine,ephedrine solution was injected into abdominal cavity of mice and ephedrine-hepatic-injury models were built.Meanwhile,Angelica Sinensis polysaccharide solution was injected into mice to protect the liver and some indices were measured,including mice weight,liver weight,structure of hepatic tissues,blood biochemical index,the activities of antioxidant enzymes and expression of specific protein related.Methods:72 2-week-old mice were chose and randomly divided into 4 groups:natural control,experiment control,ASP1,and ASP2.The experiment control group and ASP group were injected with 0.2 ml ephedrine with concentration 0.4 g/L twice per day for 15 days.The natural control group was injected with saline,the amount of which was the same as the ephedrine solution.After ephedrine solution was injected into mice as the experiment control group and ASP groups, 0.3 ml Angelica Sinensis polysaccharide solution with concentration with 12.5g/L and 25.0g/L was irrigated injected into the stomach of mice as ASP groups,respectively, while the same saline was irrigated into the stomach of mice as the natural control group and experiment control group.6 mice in every group were injected for 5,10,and15 days and then were killed to observe the change of weight of body and liver.Colorimetry was used to check the activity of γ-GT,AKP,ALT and AST in mice plasma and SOD,GSH-PX and T-AOC in mice liver and the change of MDA amount in the liver.Light-microscope and immunohistochemistry were used to observe the change of liver organization and the expressing change of NF-κB,TNF-α,Bax,Caspase-3and TGF-β1 in the hepatic tissues,respectively.Results:1. The changes of the body weight and liver weight and the radio(Wliver/Wbody)of mice:compared with normal contrast group,the body weight and liver weight of mice as experiment control groups was reduce. But after Angelica Sinensis polysaccharide solution were injected into these mice, compared with experimentcontrol group,the weight of mice as ASP1 and ASP2 was rising, but lower natural control group;in the different stages,the radio(Wliver/Wbody) of mice as experiment control group was higher than that of normal contrast group. Compared with the experiment control group,the radio of mice as ASP groups were reduced.2. The change of the structure of mice liver:the structure of mice as natural control group was complete and their lobular and liver cells were seen clearly,while hepatic tissues mice as experiment control group was injured,their cell was chaos and loose,liver sinusoid was expanded and endotheliocyte was falling.In every stage,the damage of hepatic tissues of mice as ASP groups was lighter than that of experiment control groups.For mice in the experiment control contrast group,their liver lobules were intact,liver sinusoid expanded lightly,liver cells was slightly swollen, inflammatory cells infiltration significantly reduced and central vein and hepatic sinusoidal decreased. For mice in ASP2 groups, lobular contours were observed clearly,but liver cell damage couldn’t be seen.3. The change of activities of plasma γ-GT,AKP,ALT and AST for mice liver:compared with natural control group,the activities of plasma γ-GT,AKP,ALT,AST for experiment control group mice were rising with the increase of ephedrine-injected time,while those of groups ASP1,ASP2 were reduced.4. The change of activities of antioxidant enzymes of mice liver:compared with natural control group,the activities of SOD,GSH-PX and T-AOC were significantly reduced and the content of MDA in liver tissue is increased significantly with the increase of ephedrine-injected time as the experiment control group, while the activities of SOD,GSH-PX and T-AOC of groups ASP1 and ASP2 were reduced and the content of MDA is significantly decreased.5. The change of expression of protein(including NF-κB, TNF-α, Bax,Caspase-3,TGF-β1)for mice liver:the results of immunohistochemistry represented that the positive expressing amount of NF-κB,TNF-α,Bax,Caspase-3 and TGF-β1for experiment control group was extremely low and stained color was light,while that of natural control group was rising with the increase of ephedrine-injected time and stained color was dark.Furthermore,the expression amount of NF-κB,TNF-α,Bax,Caspase-3 and TGF-β1 for groups ASP1 and ASP2 was low and their positiveexpressing amount was reduced significantly.Conclusion:1. The Angelica Sinensis polysaccharide had an effect on the weight and ratio(Wbody/Wliver) of mice liver injured by ephedrine. Ephedrine was a medicine which can mediate central nervous system.The medicine could excite hypothalamic satiety center,raise the sense of satiety,prevent the activity of feeding center,and give rise to nerve excitability and increase the activity. This process consumed a mounting deal of energy, decreasing the mice weight and speeding up the decomposition of glycogen.In addition,with the increasing of ephedrine-injected time,mice liver was damaged and its weight was decreased.But the ratio(Wbody/Wliver)of mice was increased. However, the Angelica polysaccharide could replenish consuming energy,reduced the damage to mice liver by ephedrine,which increased the weight of mice body and liver,the ratio(Wbody/Wliver) of mice was decreased.2. The Angelica Sinensis polysaccharide had an effect on mice hepatic tissues injured by ephedrine.After ephedrine was injected into mice,the accumulation of its metabolic remains in the liver did damage to hepatic tissues to different extent.As time increasing,the damage of hepatic tissues became increasingly strong,while Angelica Sinensis polysaccharides could make macrophage activated to remove the damaged cells,which contributed to multiplication of new liver cells and updating liver protein.This alleviated damage to the mice liver and liver cell apoptosis was significantly reduced.3. The Angelica Sinensis polysaccharide had an effect on the activities of plasma(including γ-GT,AKP,ALT and AST) injured by ephedrine.After ephedrine was injected into mice,the activities of the plasma were rising.The reason for it might be that ephedrine absorbed by liver cells gave rise to the inflammation of liver cells and liver sinusoid eclasis.This led to some damage to blood circulation in the liver.As ephedrine-absorbed time increasing, the activities of the plasma were rising significantly.However,the Angelica Sinensis polysaccharide could contribute to substance metabolism in the liver cell,increase the blood flow in the liver,mediate blood-sugar balances,effectively inhibit the peroxidation of cytolipin and keep the integrity of cell membrane.Therefore,the process reduced the activities of γ-GT,AKP,ALT and AST.4. The Angelica Sinensis polysaccharide solution affected activities of antioxidant enzymes in liver of mice injured by ephedrine.After ephedrine was injected into mice, it could be oxidized to activated oxygen species by cytochrome P450,producing multiple free radicals. Lipid peroxidation occurred. The activities of antioxidant enzymes SOD,GSH-PX and T-AOC were reduced significantly,while the content of MDA was rising. As ephedrine-absorbed time increasing,time effect occurred.However,Angelica Sinensis polysaccharide had a strong ability to remove reactive oxygen free radicals,inhibit lipid peroxidation of liver cells and effectively reduce liver cell damage.The activities of antioxidant enzymes SOD,GSH-PX and T-AOC activity were rising and the content of MDA was decreased.5. The Angelica Sinensis polysaccharide solution affected the expression of NF-κB, TNF-α, Bax, Caspase-3 and TGF-β1 in liver of mice injured by ephedrine.Compared with normal control group,the expression of proteins NF-κB、TNF-α,Bax,Caspase-3 and TGF-β1 in the experience control group was significantly rising,which indicated that ephedrine could make liver cells inflammatory,and that the apoptosis of liver cells was increased. However, the Angelica Sinensis polysaccharide could alleviate inflammation in the liver cells, and inhibit the expression of proteins NF-κB,TNF-α,Bax,Caspase-3,TGF-β1 and hepatocyte apoptosis,namely,the Angelica Sinensis polysaccharide protected mice liver.