| Research objectives:Based on previous studies Erzhi Pill,this experiment further separated and extracted Ligustrum ligustrum and Ecliptae Herba in Erzhi Pill,and identified their active Parts by ultraviolet spectrophotometry.By observing the effects on behavior and related indexes of aging model rats,statistics and analysis of various data,to study the anti-aging mechanism of each active part of Erzhi Pill.Research methods:1.70 SPF male SD rat were selected as experimental animals and randomly divided into 7 groups with 10 rats in each group.Blank control group,model control group,flavonoid active parts group,polysaccharide active parts group,polysaccharide active parts+flavonoid active parts+triterpenoid active parts group,Erzhi Pill decoction group.2.Drug separation and extraction: By reflux method with ethanol and water extract the active parts of triterpenes,flavonoids and polysaccharides in Erzhi Pill,and by macroporous resin adsorption method separated it.3.Modeling method: Subacute aging rat model induced by intraperitoneal injection of D-galactose.5g D-galactose was dissolved in 500 m L of 9%sodium chloride solution at an injection dose of 100 mg/(kg·d).The blank group is injected with an equal volume of normal saline.Weighed once a weekand,adjust the injection dose according to the last body weight.The modeling time for 56 days.4.Administration method: After the modeling was completed,administration group were given intragastric administration.The blank group and model group were given intragastric administration with equal volume of normal saline at the dose of 1ml/100g/d.The dose of active parts of Erzhi Pill was converted into the human dose and the body surface area of rats by intragastric administration,that is,10ml/kg/d,equivalent to 3g/kg of raw medicinal materials;the intragastric dose of Erzhi Pill decoction group was3.24g/kg/d,weighed once a week,and the intragastric dose was adjusted according to the last weighing,and administered continuously for 42 days.5.Index detection: Observe and record rats evading latency and crossing the platform.Used Elisa to detect the content of AMPK in liver and kidney tissue,the content of FOXO3 and SIRT1 in heart tissue,and the content of MLT in serum.6.The data is processed using SPSS22.0 statistical software and Excel table.Research results:1.The rat behavior experiment: Compared with the blank group,the escape latency time of rats in the model group was significantly increased,and the times of crossing the platform was significantly decreased,with significant difference(P<0.01);compared with model group,the escape latency time of rats in each active site administration group was significantly reduced,and The Times of crossing the platform was significantly increased,with significant differences(P<0.01).2.AMPK content in liver of rats: Compared with blank group,AMPK content in liver of rats in model group was significantly decreased(P<0.01);compared with model group,AMPK content in liver of triterpene group was significantly increased(P<0.01),and AMPK content in liver of flavonoid group,polysaccharide group and Erzhi Pill decoction group was significantly increased(P<0.05).Compared with Erzhi Pill decoction group,there was no difference in the administration of active parts of Erzhi Pill group(P>0.05).3.AMPK content in kidney of rats: Compared with blank group,AMPK content in kidney of rats in model group decreased significantly(P<0.01);compared with model group,the expression of AMPK in kidney of Erzhi Pill decoction group,flavonoid group,polysaccharide group and triterpene group was significantly increased(P<0.01),and the expression of AMPK in kidney of combined administration group was significantly increased(P<0.05).Compared with Erzhi Pill decoction group,there was no difference in the administration of active parts of Erzhi Pill group(P>0.05).4.FOXO3 protein expression in rats: Compared with blank group,FOXO3 protein expression in model group was significantly decreased(P<0.01);compared with model group,the expression level of FOXO3 protein in Erzhi Pill decoction group and active site administration groups increased significantly(P<0.01);compared with Erzhi Pill decoction group,there was significant difference in the active site combined administration group(P<0.01),but no difference in the other administration groups(P>0.05).5.SIRT1 protein expression in rat heart: Compared with blank group,SIRT1 protein expression in model group was significantly decreased(P<0.01);compared with model group,SIRT1 expression was increased in triterpene group(P<0.05),and SIRT1 expression was significantly increased in other active parts administration groups and water decoction administration groups(P<0.01).Compared with Erzhi Pill decoction group,flavonoid group and polysaccharide group had differences(P<0.05),combined administration group had significant differences(P<0.01),triterpenoid part administration group had no differences(P>0.05).6.Serum MLT content of rats: Compared with blank group,the serum MLT content of rats in model group was significantly decreased,with significant difference(P<0.01);compared with model group,serum MLT content in triterpene group,flavonoid group and Erzhi Pill decoction group was significantly increased(P<0.01),while serum MLT content in polysaccharide group and combined parts group was significantly increased(P<0.05).Compared with Erzhi Pill decoction group,there was no difference in the results of different active parts of Erzhi Pill group(P>0.05).Conclusion:1.The active parts of triterpenes,flavonoids and polysaccharides were extracted from Ligustrum ligustrum and Myrtaceum mongolicum in Erzhi Pill.2.The decoction of Erzhi Pill and its active components can improve the general state and learning ability of aging rats,regulate the activity of AMPK in liver and kidney,the expression levels of FOXO3 and SIRT1 in heart tissue,and regulate the content of serum MLT.3.It was found that in terms of affecting AMPK activity in liver and kidney and serum MLT content,the efficacy of each active part of Erzhi pill was equivalent to that of water decoction group;In terms of affecting FOXO3 protein expression in the heart,the active site administration group was better than the water administration group.In terms of affecting SIRT1 protein expression in heart,flavonoid group and polysaccharide group were superior to water decoction group,and combined site administration group was significantly superior to water decoction group.4.The active parts of Erzhi Pill can improve the activity of antioxidant enzymes,reduce the content of free radicals,inhibit oxidative stress,and delay aging. |
PDF Full Text Request