目的 探討孕酮對(duì)皮質(zhì)酮誘導(dǎo)的PC12抑郁癥細(xì)胞模型的保護(hù)作用及其潛在機(jī)制。方法 CCK-8法檢測(cè)細(xì)胞存活率篩選皮質(zhì)酮(100、200、300、400、500、600μmol·L^-1)作用于PC12細(xì)胞構(gòu)建抑郁癥細(xì)胞模型；造模后，CCK-8法篩選孕酮(1、5、10、20、40、60、80μmol·L^-1)的治療濃度；同時(shí)設(shè)定孕酮核受體（n PR）特異性抑制劑RU486和孕酮受體膜組分1（PGRMC1）特異性抑制劑AG205聯(lián)合孕酮(10μmol·L^-1)處理組，加藥后孵育36 h。流式細(xì)胞術(shù)檢測(cè)凋亡情況；DCFHDA探針法測(cè)定細(xì)胞內(nèi)活性氧（ROS）水平；鈣離子熒光探針Fluo-4 AM法測(cè)定鈣離子濃度；試劑盒法檢測(cè)乳酸脫氫酶（LDH）釋放情況；JC-1探針法檢測(cè)線粒體膜電位，并采用Western blotting法分析凋亡相關(guān)蛋白的表達(dá)。結(jié)果 PC12抑郁癥細(xì)胞模型采用皮質(zhì)酮300μmol·L^-1制備，孕酮10μmol·L^-1作用效果較好；與模型組相比，孕酮顯著降低細(xì)胞內(nèi)ROS水平和鈣離子濃度（P<0.01），減少LDH釋放以及穩(wěn)定線粒體膜電位（P<0.01），提高Bcl-2蛋白的表達(dá)（P<0.01），降低Bax、Cyt C、Caspase-3蛋白的表達(dá)（P<0.05），從而減少細(xì)胞凋亡（P<0.01），提升細(xì)胞活力（P<0.01）；且RU486和AG205均能抑制孕酮對(duì)抑郁癥細(xì)胞模型的保護(hù)作用（P<0.05、0.01）。結(jié)論 孕酮能通過n PR以及PGRMC1減輕皮質(zhì)酮誘導(dǎo)的PC12抑郁癥細(xì)胞模型氧化應(yīng)激反應(yīng)以及細(xì)胞內(nèi)鈣超載，進(jìn)而發(fā)揮神經(jīng)保護(hù)作用。

Objective To explore the protective effect of progesterone on the corticosterone-induced PC12 depression cell model and its potential mechanism. Methods The CCK8 method was used to detect cell viability to screen the concentration of corticosterone(100, 200, 300, 400, 500, 600 μmol·L^-1) for the construction of the depression cell model in PC12 cells; After modeling, the CCK8 method was used to screen the therapeutic concentration of progesterone(1, 5, 10, 20, 40, 60, 80 μmol·L^-1); At the same time, the progesterone nuclear receptor（nPR） specific inhibitor RU486 and the progesterone receptor membrane component 1（PGRMC1） specific inhibitor AG205 combined with progesterone(10 μmol·L^-1) treatment groups were set up, and the cells were incubated for 36 h after drug addition. Apoptosis was detected by flow cytometry; intracellular reactive oxygen species（ROS） levels were detected by the DCFH-DA probe method, intracellular calcium ion concentration was detected by the calcium ion fluorescent probe Fluo-4 AM method; lactate dehydrogenase（LDH） release was detected by the kit method, and mitochondrial membrane potential was detected by the JC-1 probe method, and the expression of apoptosis-related proteins was analyzed by Western blotting. Results The PC12 depression cell model was prepared with 300 μmol·L^-1 corticosterone, and 10 μmol·L^-1 progesterone had a better effect; Compared with the model group, progesterone significantly reduced intracellular ROS levels and calcium ion concentration（P＜0.01）, reduced LDH release and stabilized mitochondrial membrane potential（P＜0.01）, increased the expression of Bcl-2 protein（P＜0.01）, decreased the expression of Bax, Cyt C, and Caspase-3 proteins（P＜0.05）, thereby reducing cell apoptosis（P＜0.01） and enhancing cell viability（P＜0.01）; and both RU486 and AG205 could inhibit the protective effect of progesterone on the depression cell model（P＜0.05, 0.01）. Conclusion Progesterone exerts neuroprotective effects by alleviating CORT-induced oxidative stress and intracellular calcium overload in the PC12 depression cell model through nPR and PGRMC1 pathways.

R965

河北省省屬高校基本科研業(yè)務(wù)費(fèi)自然科學(xué)研究計(jì)劃項(xiàng)目(JYT2021001); 河北省研究生創(chuàng)新資助項(xiàng)目(CXZZSS2024129); 河北北方學(xué)院校級(jí)科研項(xiàng)目(XJ2023034)