目的 應(yīng)用斑馬魚模型評價紫河車凍干粉水提物（PHFAE）抗衰老的功效。方法 將健康產(chǎn)婦新鮮胎盤冷凍干燥后碾磨為粉，并使用蒸餾水進行提取，制成紫河車凍干粉水提物（PHFAE）。發(fā)育8 h （8 hpf）的AB系斑馬魚使用PHFAE（0、1、10、25 μg·mL^-1）連續(xù)作用72 h，對其體內(nèi)β-半乳糖苷酶進行染色。隨后將AB系與皮膚熒光轉(zhuǎn)基因斑馬魚均隨機分成對照組、模型組和PHFAE （1、10、25 μg·mL^-1）組，對照組給予培養(yǎng)水，模型組給予0.5 mmol·L^-1 H₂O₂溶液制備氧化損傷模型，PHFAE組同時給予H₂O₂與不同濃度PHFAE，連續(xù)作用72 h后，測定皮膚表面熒光斑點的數(shù)量；試劑盒法檢測魚體內(nèi)活性氧（ROS）含量；實時熒光定量PCR（qRT-PCR）法檢測ampk、mdm2、Cu/Zn-sod和Mn-sod的mRNA表達變化。結(jié)果 與對照組比較，1、10、25 μg·mL^-1的PHFAE組斑馬魚體內(nèi)β-半乳糖苷酶的活性均顯著降低（P＜0.01）；與模型組比較，1、10、25 μg·mL^-1的PHFAE顯著抑制H₂O₂誘導的角質(zhì)細胞的死亡（P＜0.01），明顯抑制ROS的積聚，顯著升高ampk、mdm2、Cu/Zn-sod和Mn-sod的mRNA表達（P＜0.01）。結(jié)論 PHFAE通過調(diào)控ampk、mdm2、Cu/Zn-sod和Mn-sod基因的表達，提高了機體的抗氧化能力，進而發(fā)揮抗衰老之功效。

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 25 μg·mL^-1) for 72 h. Followingly, zebrafish β-galactosidase was stained. Then both AB and Tg (krt4: NTR-hKikGR) strain zebrafish were divided into control, model and PHFAE (0, 1, 10 and 25 μg·mL^-1) groups. The control group was given zebrafish culture water, the model group was given 0.5 mmol·L^-1 H₂O₂ solution to prepare oxidative damage model, and the administration group was given H₂O₂ 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 of β-galactosidase in PHFAE group of 1, 10, 25 μg·mL^-1 was significantly reduced (P < 0.01). Compared to model group, PHFAE of 1, 10, 25 μg·mL^-1 significantly inhibited H₂O₂ 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.

R285.5

山東省科技型中小企業(yè)創(chuàng)新能力提升工程項目（2022TSGC1036）；山東省自然科學基金面上項目（ZR2022MD102）；齊魯工業(yè)大學（山東省科學院）科教產(chǎn)融合創(chuàng)新試點工程項目（2022PX061，2022PX021）。