| Objective : To explore the protection of chlorogenic acid from Eucommia ulmoides Oliver on hepatic ischemia-reperfusion injury in rats and mechanism,to provide theoretical foundation for clinical application in hepatic ischemia reperfusion injury.Methods: 48 male SD mice(160g-200g)were randomly divided into 6 group including control group(SHAM),ischemia reperfusion group(IR)and ulinastain group,chlorogenic acid low does group,medium dose group and high dose group.After each group of mice were adapted to the new environment for a week,lavage for 10 days,where the dosage was20ml/kg.d.The mice in the operation group,the normal group and ulinastatin group,were given normal saline to fill the stomach.In addition,the ulinastatin group was given normal saline to fill the stomach.In addition,the ulinastatin group was given 30000U/kg.ulinastin intraperitioneal injection 30 minutes before surgery.The concentrations of the chlorogenic acid low dose group,medium dose group and high dose group were blocked,causing 70%of hepatic tissue ischemia in the remaining groups.After 1 hour of ischemia in each group,the venous blood and liver tissue were removed after 4 hour of perfusion.The serum transaminase level was detected and HE staining was used to evaluate its protective effect pathologically.the activity of SOD in liver tissue TNF-?,IL-? levels.The expression of HMGB1 in liver tissue was detected by ELISA and Western Blot,and the protective mechanism of chorogenic acid from Eucommia ulmoides Oliver was discussed.Results:(1)Liver histopathological examination.The pathological section HE staining was visible under the light microscope.In SHAM group,liver cells were normal and liver cells were arranged without obvious changes.In IR group,the hepatic cord in liver tissue was disorganized,hepatocyte edema was obvious,the hepatic sinus narrowed and a lot of inflammatory cells infiltrated and so on.Compared with the operation group,the damage of chlorogenic acid pretreatment group and ulinastatin group was significantly reduced and the effect was the most significant in chlorogenic acid high dose group.(2)Serum transaminase detection.After the pretreatment of chlorogenic acid and ustodin from eucommia ulmoides,the levels of aminotransferase of mice were lower than that in IR group(P<0.05),among them,the effect was the most significant in the chlorogenic acid high dose group.(3)MDA level of liver homogenate.The MDA content of IR group increased significantly.The MDA concentration in each group was lower than that in IR group(P<0.05),in which the effect in the chlorogenic acid high dose group was more obvious.The MDA content in rats after pretreatment of ulinastatin was lower than that in IR group.However,the effect was weaker than that of chlorogenic acid high dose group(P< 0.05).(4)SOD activity of liver homogenate.The SOD activity in SHAM group was significantly higher than that in IR group(P<0.05).The SOD content in the chlorogenic acid groups was also higher than that of IR group(P<0.05),among that chlorogenic acid high group the most.The activity of SOD in ulinastatin group was higher than that of the IR group,but was weaker than that of chlorogenic acid high dose group(P <0.05).(5)Hepatic homogenate IL beta level.The level of IL-1? in IR group was higher than that of other groups.In the chlorogenic acid group of pretreatment and the ulinastatin group,the expression of IL-1? was lower than that in the IR group but higher than that in the SHAM group(P < 0.05).Among them,the decrease of chlorogenic acid high dose group was more significant.(6)liver homogenate TNF-?level,The TNF-?level in IR group was higher than the other groups.The expression of TNF-?in the pretreatment group of chlorogenic acid and ulinastatin group was lower than that in the IR group but higher than that in the SHAM group(P < 0.05),and the decrease was more significant in the chlorogenic acid high dose group.(7)Liver homogenate HMGB1 expression level.ELISA and Western Blot results showed that HMGB-1 expression in IR group was higher than that in other groups.Chlorogenic acid and ulinastatin can reduced HMGB-1 expression(P< 0.05),and chlorogenic acid high dose group effect was superior to Ulinastatin group.Conclusion:(1)The chlorogenic acid from eucommia ulmoides could protect HIRI in rats by anti-inflammatory and anti-oxidation effects.chlorogenic acid high dose group the protective effect is better than that of ulinastatin.(2)The protective mechanism of chlorogenic acid from eucommia ulmoides on HIRI in rats may be related to the release and expression of HMGB1,and the specific molecular mechanism needs research in depth. |
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