鼠尾草酸对H2O2诱导的早熟性细胞衰老和D-半乳糖诱导的衰老模型小鼠的保护作用  

Protective effects of carnosic acid against aging in a premature cellular senescence model and in a D-galactose induced mouse model

作　　者：苏慧丽[1] 暴一众 张婧[1] 徐小刚 张忠山[2] 万晓青[1] 毛根祥[1] SU Hui-li;BAO Yi-zhong;ZHANG Jing;XU Xiao-gang;ZHANG Zhong-shan;WAN Xiao-qing;MAO Gen-xiang(Zhejiang Hospital,Hangzhou 310013,China;School of Life Sciences,Huzhou University,Huzhou 313000,China)

机构地区：[1]浙江医院,浙江杭州310013 [2]湖州师范学院生命科学学院,浙江湖州313000

出　　处：《药学学报》2020年第5期915-921,共7页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81771520,31700307);浙江省医药卫生平台计划(学科带头人)(2015DTA001);浙江省中医药科学研究基金项目(2015ZA002);浙江省中医药优秀青年人才基金项目(2020ZQ002)。

摘　　要：研究鼠尾草酸(carnosic acid, CA)延缓早熟性细胞衰老和对D-半乳糖(D-gal)诱导小鼠衰老的保护作用和相关机制。采用人胚肺二倍体成纤维2BS细胞系,应用200μmol·L^-1H2O2建立早熟性细胞衰老模型,观察CA对衰老相关β-半乳糖苷酶(SA-β-Gal)活性和衰老相关蛋白p53、p21和p16的影响。动物实验获得浙江医院伦理委员会批准。采用雄性C57 BL/6J小鼠连续8周皮下注射D-gal (100 mg·kg^-1·d^-1)构建小鼠衰老模型,给予5和10 mg·kg^-1·d^-1CA进行干预, Morris水迷宫实验观察小鼠空间记忆能力变化、测定小鼠血清及海马组织中脂质过氧化产物丙二醛(MDA)、总超氧化物歧化酶(T-SOD)水平、晚期糖基化终末产物(AGEs)、炎症因子白介素-6 (IL-6)和肿瘤坏死因子α(TNFα)水平,并检测海马内衰老相关分子的蛋白表达变化。结果显示, H2O2处理的2BS细胞SA-β-Gal染色阳性率接近95%,而CA干预组为35%左右,并显著降低H2O2诱导的衰老相关分子p53、p21和p16蛋白水平。在D-gal小鼠模型中, CA干预明显改善了小鼠空间记忆能力,并显著抑制AGEs生成、氧化应激和炎症因子水平,同时减少小鼠海马中衰老相关p53、p21和p16蛋白表达。研究表明, CA可改善氧化应激诱导的2BS细胞早熟型衰老,并可以明显改善小鼠空间记忆能力,该作用与其抗氧化、抗炎及抑制非酶糖基化作用相关。This study aimed to investigate the effect and possible mechanism of carnosic acid(CA) on delaying aging. The effects of CA on senescence-related β-galactosidase(SA-β-Gal) activity and expressions of p53, p21 and p16 were evaluated by an oxidative challenge induced premature 2 BS cell senescence model. Meanwhile,the animal experiment was approved by the Ethics Committee of Zhejiang Hospital. Male C57 BL/6 J mice were injected with 100 mg·kg^-1·d^-1 D-galactose(D-gal) for 8 weeks to establish an aging model in vivo, and CA at 5 and10 mg·kg^-1·d^-1 were given ig administration at the same time. Morris water maze test was used to test the spatial memory ability. Then the serum and tissue samples were collected for the detections of malondialdehyde(MDA),total superoxide dismutase(T-SOD), interleukin-6(IL-6), tumor necrosis factor α(TNFα) and advanced glycation end products(AGEs) as well as the protein expression of p53, p21 and p16 in hippocampus of brain. The results showed that H2O2 induced increment of SA-β-Gal activity(95%) was prevented by CA treatment(35%) and the enhanced protein expressions of p53, p21 and p16 in H2O2 exposed 2 BS cells were alleviated by CA treatment,suggesting a potent protective role of CA against premature senescence induced by oxidative challenge. For in vivo study, D-gal induced declined spatial memory ability was partly reversed by CA administration. Besides, the serum and cerebral levels of MDA, IL-6, TNFα and AGEs were attenuated by CA treatment when compared to those in model mice. And the protein expressions of p53, p21 and p16 in mice hippocampus were suppressed by CA in D-gal treated mice. Taken together, our results showed that CA protects premature senescence induced by oxidative stress and D-gal, which is related to its antioxidative, antiinflammatory roles and inhibition on non-enzymatic glycosylation.

关 键 词：鼠尾草酸 衰老 D-半乳糖 非酶糖基化 氧化应激 

分 类 号：R965[医药卫生—药理学]