宁夏无果枸杞芽水提物对老年小鼠心肌细胞抗氧化及凋亡蛋白表达的影响

摘要

目的 观察宁夏无果枸杞芽水提物对老年小鼠心肌细胞抗氧化能力及凋亡相关蛋白表达的影响.方法 将13个月龄自然衰老的C57BL/6J小鼠分为老年对照组和AEFLS低剂量组(AEFLS1组)、AEFLS中剂量组(AEFLS2组)、AEFLS高剂量组(AEFLS3组).AEFLS1组、AEFLS2组、AEFLS3组分别给予5、10、20 mg/kg的AEFLS灌胃,老年对照组灌胃生理盐水,连续8周.分别用黄嘌呤氧化酶法和硫代巴比妥酸法测定心脏组织SOD和MDA的水平;Western-blot技术和免疫组织化学方法检测小鼠心肌组织Bcl-2、Bax和Capase-3的表达.结果 与老年对照组比较,AEFLS2和AEFLS3组MDA水平降低,SOD活性升高,差异有统计学意义(P＜0.01).Western-blot结果:与老年对照组比较,AEFLS2和AEFLS3组的Bcl-2蛋白光密度值升高,Bax和Capase-3蛋白光密度值降低(P＜0.01);免疫组化结果:与老年对照组比较,AEFLS2和AEFLS3组Bcl-2蛋白免疫阳性表达均升高,Bax和Capase-3蛋白免疫阳性表达降低(P＜0.01).结论 中、高剂量的AEFLS能提高衰老小鼠的心肌组织抗氧化能力,上调Bcl-2的表达,下调Capase-3和Bax的表达,发挥抗心肌细胞凋亡的作用.%Objective To observe the effect of Ningxia aqueous extracts of fruitless lycium sprout (AEFLS) on cardiomyo cyte antioxidation and apoptosis-related protein expression in aging mice.Methods The natural aging C57BL/6J mice with 13 months old were randomly divided into aged control group,AEFLS low dose group (AEFLS1),AEFLS middle dose group (AEFLS2)and AEFLS high dose group(AEFLS3).The AEFLS1,AEFLS2 and AEFLS3 groups were respectively given with 5,10,20 mg/kg AEFLS gavage,while the aged control group was given with the normal saline gavage,for continuous8 weeks.The xan thine oxidase assay and thiobarbituric acid method were used for the determination of SOD and MDA in heart tissues.Western-blot and immunohistochemical method were used to detect the expressions of Bcl-2,Bax and Capase-3 in heart tissue.Results Compared with the aged control group,the MDA level in the AEFLS2 and AEFLS3 groups was decreased,while the SOD activity was increased,the difference was statistically significant (P＜0.01).The Western-blot result showed that compared with the aged control group,the optical density value of Bcl-2 in the AEFLS2 and AEFLS3 groups was increased,but the optical density values of Bax and Capase-3 were decreased (P＜0.01);the immunohistochemical results showed that compared with the aged control group,the immunopositive(IP) expressions of Bcl-2 protein in heart tissues in the AEFLS2 and AEFLS3 groups were increased (P＜0.01),while the IP expressions of Bax and Capase-3 were decreased (P＜0.01).Conclusion Middle and high doses of AEFLS can increase the antioxidative ability of myocardial tissue,up-regulates the Bcl-2 expression,down-regulates the Bax and Capase-3 expressions and plays anti-cardiomyocyte apoptotic role.