| Research purposes:Based on the previous researches on Erzhi Pill,Ligustrum lucidum and Eclipta chinensis in Erzhi Pill were further separated and extracted,and the active parts were preliminarily clarified.By observing the effect of each active part of Erzhi Pill on the pathological state and related indexes of aging model rats,the anti-aging effect of the active parts of Erzhi Pill was discussed.Research methods:1.70 SPF male SD rats were selected as experimental animals and randomly divided into 7 groups with 10 rats in each group.Flavonoid active parts group(hereinafter referred to as flavonoid group),polysaccharide active parts group(hereinafter referred to as polysaccharide group),polysaccharide active parts+flavonoid active parts+triterpenoid active parts group(hereinafter referred to as combined group),Erzhi Pill decoction group(hereinafter referred to as the decoction group).2.Drug separation and extraction:The triterpenes(glycosides),phenols(flavonoids)and polysaccharides in Erzhi Pill(Ligustrum lucidum and Eclipta chinensis)were extracted by ethanol and water double-solvent reflux method and macroporous resin adsorption method respectively.active parts.3.Modeling method:A rat subacute aging model was induced by intraperitoneal injection of D-galactose.5g of D-galactose was dissolved in500 m L of 0.9% sodium chloride solution,and the injection dose was 100mg/(kg·d)(ie,0.01 m L per gram of body weight).The blank group was injected with an equal volume of normal saline.Weigh once a week,and adjust the injection dose according to the last body weight.The modeling time lasted 56days(8 weeks).4.Administration method:After successful modeling,each administration group was given intragastric administration from the 9th week,and the blank group and the model group were respectively intragastrically administered with an equal volume of normal saline at a dose of 1ml/100g/d.Each active parts group of Erzhi Pill was administered by intragastric administration according to the human dose and the body surface area of rats,that is,10ml/kg/d,and the administered dose was equivalent to 6g/kg of raw medicinal materials.The intragastric administration of Erzhi Pill decoction group was 3.24g/kg/d,weighed once a week,adjusted the intragastric dose according to the last weighing,and administered continuously for 42 days(6weeks).5.Index detection:The escape latency,average swimming speed and the number of crossing the platform of rats were observed.Enzyme-linked immunosorbent assay(ELISA)was used to detect the contents of SOD and MDA in liver and spleen tissues,HYP content in skin tissue,Klotho protein expression and 8-OHd G content in serum.6.The data were processed using SPSS23.0 statistical software and Excel table data.Research result:1.Behavioral expression of rats:Compared with the blank group,the escape latency of rats in the model group was significantly increased,the average swimming speed was significantly decreased,and the number of crossing platforms was significantly decreased,with significant differences(P<0.01);In comparison,the escape latency of the rats in Erzhi Pill and each active parts administration group was significantly reduced,with a significant difference(P<0.01);the average swimming speed was significantly increased,and the difference in the flavonoid group was not significant(P>0.05);The number of platforms increased significantly,with a significant difference(P<0.01).2.SOD content in rats:Compared with the blank group,the SOD activity of the rats in the model group was significantly reduced,and there was a significant difference(P<0.01).The vitality was significantly improved,with significant difference(P<0.01),and there was no difference in the flavonoid group(P>0.05).3.MDA content in rats:Compared with the blank group,the MDA activity of the rats in the model group was significantly increased,and there was a significant difference(P<0.01).The activities of MDA were significantly decreased,with a significant difference(P<0.01).4.8-OHd G content in rats:Compared with the blank group,the content of8-hydroxydeoxyguanosine(8-OHd G)in the model group was significantly increased,with a significant difference(P<0.01);The content of 8-OHd G in Erzhi Pill and each active parts administration group was significantly decreased,with difference(P<0.05).5.The content of HYP in rats:Compared with the blank group,the content of hydroxyproline(HYP)in the model group was significantly reduced(P<0.01);compared with the model group,the content of hydroxyproline(Hyp)in Erzhi Pill and its active parts increased significantly(P<0.01).6.Klotho protein expression in rats:Compared with the blank group,the expression level of Klotho protein in the model group was significantly decreased(P<0.01);compared with the model group,the Klotho expression level in the combined group was increased,with a difference The expression level of Klotho was significantly different between the other active partss and the decoction group(P<0.01).Inconclusion:1.Erzhi Pill are separated and extracted to obtain active parts of triterpenes,flavonoids and polysaccharides,and their identification and designation are carried out.2.Erzhi Pill and various active parts can improve the general state and learning and memory ability of aging model rats,regulate the content of SOD and MDA in liver and spleen tissue,the content of HYP in skin tissue,and the activity of 8-OHd G in serum and Klotho.protein expression.3.Through the analysis of the experimental data and the comprehensive evaluation,it is found that the active parts of Erzhi Pill have the same anti-aging effect as the decoction.Among them,the effect of the active site combination group was better than that of the decoction group in terms of Klotho protein expression and 8-OHd G activity.4.The various active parts of Erzhi Pill may play the purpose of delaying aging by inhibiting the oxidative stress of the body,increasing the activity of the body’s antioxidant enzymes,and reducing the degree of free radicals and DNA oxidative damage. |
PDF Full Text Request