目的 明確人源胎盤凍干粉是否具有抗利血平誘發(fā)斑馬魚抑郁癥的活性，比較不同提取方法胎盤凍干粉抗抑郁活性并探索其作用機制。方法 將6月齡野生型斑馬魚AB隨機分為8組：對照組、模型組、丁螺環(huán)酮（陽性藥，3mg·L^-1）組和胎盤凍干粉酶解、水提、50%醇提、醇提、二氯甲烷提（均為5mg·L^-1）組，每組12條。除對照組外，各組以利血平處理20min造模，而后更換相應(yīng)含藥養(yǎng)魚用水，至14d，每天換水。14d時用Zebrabox斑馬魚行為分析儀對各組斑馬魚進行新型水槽測試，分析其行為變化。收集魚腦，實時熒光定量PCR（qRT-PCR）法檢測抑郁、神經(jīng)調(diào)控相關(guān)基因（bdnf、pomc、hcrt、mao、nf-κb、pparγ）的mRNA表達；試劑盒法檢測氧化應(yīng)激相關(guān)超氧化物歧化酶（SOD）和過氧化氫酶（CAT）活性。結(jié)果 與對照組比較，模型組斑馬魚進入新型水槽上部次數(shù)顯著減少（P＜0.001），上部游動距離及時間比例均顯著減少（P＜0.001）；與模型組比較，丁螺環(huán)酮、胎盤凍干粉酶解物組斑馬魚進入上部次數(shù)顯著增加（P＜0.05），上部游動距離及時間比例均顯著增加（P＜0.05）。與對照組比較，模型組中的bdnf、pomc、hcrt和mao基因表達顯著上調(diào)（P＜0.01、0.001），nf-κb和pparγ基因表達顯著下調(diào)（P＜0.05、0.01）；與模型組比較，經(jīng)不同提取方法胎盤凍干粉處理后bdnf、pomc和mao基因表達顯著下調(diào)（P＜0.05、0.01、0.001），酶解、水提、醇提組hcrt mRNA水平顯著降低（P＜0.05、0.001），酶解、水提組nf-κb mRNA水平和酶解組pparγ mRNA水平顯著上調(diào)（P＜0.01）。與對照組比較，模型組CAT活性顯著升高（P＜0.001），SOD活性升高，但沒有統(tǒng)計學意義；與模型組比較，經(jīng)不同提取方法胎盤凍干粉處理后CAT活性顯著降低（P＜0.001），SOD活性有不同程度的降低。結(jié)論 人源胎盤凍干粉具有抗利血平誘發(fā)斑馬魚抑郁癥的活性，且胎盤凍干粉酶解抗抑郁癥活性最好，作用機制可能與改善氧化應(yīng)激狀態(tài)有關(guān)。

Objective To determine whether human placenta freeze-dried powder has the activity of anti-depressant in zebrafish depression model induced by reserpine, as well as compare the anti-depressant activity of dried human placenta extracted by different methods and explore the underlying mechanism. Methods The 6-month-old wild-type zebrafish AB were divided into eight groups including blank control group, model group, buspirone (positive control, 3 mg·L^-1）group, dried human placenta enzymatic hydrolysate, water extract, alcohol extract, 50% alcohol water extract and dichloroethane extract (5 mg·L^-1）group, with 12 fish in each group. Among them, the model group was treated with reserpine for 20 min, and the other groups were subsequently treated with samples for 14 days. The medium was changed every day. After 14 days treatment, the novel tank assay was conducted in adult zebrafish to analyze the behavior changes of each group. The zebrafish brains were collected after behavior assays, which were used to test the expression of depression and neuromodulation related genes (bdnf、 pomc、 hcrt、 mao、 nf-κb、 pparγ) by real-time fluorescence quantitative PCR (qRT-PCR) method. In addition, the activities of anti-oxidative enzymes superoxide dismutase (SOD) and catalase (CAT) were detected by kits. Results Compared with control group, the number of zebrafish entering the upper part of the new tank was significantly reduced (P<0.001), and the proportion of swimming distance and time in the upper part was significantly decreased (P<0.001) in model group. Compared with model group, the number of zebrafish entering the upper part in buspiroone and placenta lyophilized powder enzymolysis groups was significantly increased (P<0.05), and the proportion of upper swimming distance and time was significantly increased (P<0.05). Compared with control group, bdnf, pomc, hcrt and mao gene expressions were significantly up-regulated (P<0.01, 0.001), while nf-κb and pparγ gene expressions were significantly decreased (P<0.05, 0.01) in model group. Compared with model group, the expressions of bdnf, pomc and mao genes were significantly down-regulated (P<0.05, 0.01, 0.001) after human placenta freeze-dried powder treatment with different extraction methods, and hcrt mRNA levels were significantly decreased in hydrolysate, water extract and alcohol extract groups (P<0.05, 0.001). The mRNA levels of nf-κb and pparγ of hydrolysate and water extract were significantly up-regulated (P<0.01). Compared with the control group, CAT activity in model group was significantly increased with statistical significance (P<0.001), while SOD activity was increased with no statistical significance. Compared with model group, CAT activity decreased significantly (P<0.001) and SOD activity decreased to different degrees after human placenta freeze-dried powder treatment with different extraction methods. Conclusion human placenta freeze-dried powder had the activity against depression induced by reserpine in zebrafish, among which the enzymatic hydrolysate of dried human placenta has the best anti-depressant activity. Mechanism investigation suggested that antidepressant activity may be related to the anti-oxidative action.

R285.5

濟南市“新高校20條”項目（2021GXRC106、2021GXRC111）；山東省重大科技創(chuàng)新工程（2021CXGC010507）