基于斑马鱼模型的紫河车冻干粉水提物抗衰老功效研究  

Research on anti-senescence effect of Placenta Hominis freeze-dried power aqueous extract based on zebrafish

作　　者：孙晨 洪凡真[2] 李艳胧 巴帅康 郑红宝 张云 臧传宝 付开玲 张虎 侯海荣 任季禹 SUN Chen;HONG Fanzhen;LI Yanlong;BA Shuaikang;ZHENG Hongbao;ZHANG Yun;ZANG Chuanbao;FU Kailing;ZHANG Hu;HOU Hairong;REN Jiyu(Qilu University of Technology(Shandong Academy of Sciences)Biology Institute,Jinan 250103,China;Second Hospital of Shandong University,Jinan 250033,China;Shandong Electric Power Central Hospital,Jinan 250001,China;Yinfeng Cryogenic Medical Technology Co.,Ltd.,Jinan 250000,China)

机构地区：[1]齐鲁工业大学(山东省科学院)生物研究所,山东济南250103 [2]山东大学第二医院,山东济南250033 [3]山东电力中心医院,山东济南250001 [4]银丰低温医学科技有限公司,山东济南250000

出　　处：《药物评价研究》2023年第11期2374-2379,共6页Drug Evaluation Research

基　　金：山东省科技型中小企业创新能力提升工程项目(2022TSGC1036);山东省自然科学基金面上项目(ZR2022MD102);齐鲁工业大学(山东省科学院)科教产融合创新试点工程项目(2022PX061,2022PX021)。

摘　　要：目的应用斑马鱼模型评价紫河车冻干粉水提物(PHFAE)抗衰老的功效。方法将健康产妇新鲜胎盘冷冻干燥后碾磨为粉,并使用蒸馏水进行提取,制成紫河车冻干粉水提物(PHFAE)。发育8 h(8 hpf)的AB系斑马鱼使用PHFAE(0、1、10、25μg·mL^(-1))连续作用72 h,对其体内β-半乳糖苷酶进行染色。随后将AB系与皮肤荧光转基因斑马鱼均随机分成对照组、模型组和PHFAE(1、10、25μg·mL^(-1))组,对照组给予培养水,模型组给予0.5 mmol·L^(-1)H_(2)O_(2)溶液制备氧化损伤模型,PHFAE组同时给予H_(2)O_(2)与不同浓度PHFAE,连续作用72 h后,测定皮肤表面荧光斑点的数量;试剂盒法检测鱼体内活性氧(ROS)含量;实时荧光定量PCR(qRT-PCR)法检测ampk、mdm2、Cu/Zn-sod和Mn-sod的mRNA表达变化。结果与对照组比较,1、10、25μg·mL^(-1)的PHFAE组斑马鱼体内β-半乳糖苷酶的活性均显著降低(P<0.01);与模型组比较,1、10、25μg·mL^(-1)的PHFAE显著抑制H_(2)O_(2)诱导的角质细胞的死亡(P<0.01),明显抑制ROS的积聚,显著升高ampk、mdm2、Cu/Zn-sod和Mn-sod的mRNA表达(P<0.01)。结论PHFAE通过调控ampk、mdm2、Cu/Zn-sod和Mnsod基因的表达,提高了机体的抗氧化能力,进而发挥抗衰老之功效。Objective To explore the anti-senescence effect of Placenta Hominis freeze-dried power aqueous extract(PHFAE)in vivo.Methods The fresh placentas from healthy puerperae were freeze-dried.Then,they were ground into powder,and extracted by water to prepare the water extracts from Placenta Hominis freeze-dried power.8 hpf AB strain zebrafish were treated with the different concentrations of PHFAE(0,1,10 and 25μg·mL^(-1))for 72 h.Followingly,zebrafishβ-galactosidase was stained.Then both AB and Tg(krt4:NTR-hKikGR)strain zebrafish were divided into control,model and PHFAE(0,1,10 and 25μg·mL^(-1))groups.The control group was given zebrafish culture water,the model group was given 0.5 mmol·L^(-1) H_(2)O_(2) solution to prepare oxidative damage model,and the administration group was given H_(2)O_(2) solution and different concentrations of PHFAE at the same time.After 72 h of treatment,the number of fluorescent spots on the skin surface were detected,content of reactive oxygen species were detected by kits,and the expressions of ampk,mdm2,Cu/Zn-sod and Mn-sod were detected by real-time fluorescence quantitative PCR(qRT PCR).Results Compared with control group,the activity ofβ-galactosidase in PHFAE group of 1,10,25μg·mL^(-1) was significantly reduced(P<0.01).Compared to model group,PHFAE of 1,10,25μg·mL^(-1) significantly inhibited H_(2)O_(2) induced keratinocyte death(P<0.01),significantly inhibited ROS accumulation,and significantly increased mRNA expression of ampk,mdm2,Cu/Zn-sod,and Mn-sod(P<0.01).Conclusion PHFAE can improve the antioxidant capacity of the body and play the anti-aging effect by regulating the expressions of ampk,mdm2,Cu/Zn-sod and Mn-sod genes.

关 键 词：紫河车冻干粉 水提取 抗衰老 抗氧化 斑马鱼 

分 类 号：R285.5[医药卫生—中药学]