目的 探討腦苷肌肽對(duì)缺氧缺血性腦病（HIE）新生大鼠沉默信息調(diào)節(jié)因子2相關(guān)酶1（SIRT1）/哺乳動(dòng)物雷帕霉素靶蛋白（mTOR）/p70核糖體蛋白S6激酶（p70S6K）通路及神經(jīng)元凋亡的影響。方法 采用結(jié)扎左側(cè)頸總動(dòng)脈及缺氧玻璃倉(cāng)的方法建立新生大鼠HIE模型，造模大鼠隨機(jī)分為5組：模型組、腦苷肌肽低劑量組（0.8 mg/kg）、腦苷肌肽中劑量組（1.6 mg/kg）、腦苷肌肽高劑量組（3.2 mg/kg）、尼莫地平組（10 mg/kg），每組12只，另取12只設(shè)為假手術(shù)組。分組處理后，觀察各組大鼠神經(jīng)行為活動(dòng)并進(jìn)行評(píng)分；測(cè)定各組大鼠腦組織含水量；TUNEL染色檢測(cè)各組大鼠腦皮質(zhì)神經(jīng)元凋亡情況；酶聯(lián)免疫吸附法（ELISA）檢測(cè)各組大鼠血清腫瘤壞死因子-α（TNF-α）、白細(xì)胞介素-1β（IL-1β）水平；免疫印跡法檢測(cè)各組大鼠腦組織SIRT1/mTOR/p70S6K通路相關(guān)蛋白表達(dá)情況。結(jié)果 與假手術(shù)組相比，模型組大鼠神經(jīng)功能缺損評(píng)分、腦組織含水量、腦皮質(zhì)神經(jīng)元凋亡率、血清TNF-α及IL-1β水平、腦組織SIRT1/mTOR/p70S6K通路相關(guān)蛋白p-mTOR/mTOR及p-p70S6K/p70S6K明顯升高，腦組織SIRT1表達(dá)降低（P<0.001）；與模型組相比，腦苷肌肽低、中、高劑量組及尼莫地平組大鼠神經(jīng)功能缺損評(píng)分、腦組織含水量、腦皮質(zhì)神經(jīng)元凋亡率、血清TNF-α及IL-1β水平、腦組織SIRT1/mTOR/p70S6K通路蛋白p-mTOR/mTOR及p-p70S6K/p70S6K降低，腦組織SIRT1表達(dá)升高（P<0.001），且腦苷肌肽各組呈劑量依賴性（P<0.001），腦苷肌肽高劑量組與尼莫地平組相比，大鼠各指標(biāo)差異無(wú)統(tǒng)計(jì)學(xué)意義。結(jié)論 腦苷肌肽可促使HIE新生大鼠腦組織SIRT1表達(dá)，降低mTOR、p70S6K磷酸化水平，抑制炎癥，減輕腦水腫及神經(jīng)元凋亡，修復(fù)神經(jīng)功能，改善其臨床癥狀。

Objective To investigate the effect of cattle encephalon glycoside and ignotin on sirtuin 1 (SIRT1)/mammalian target of rapamycin (mTOR)/p70 ribosomal protein S6 kinase 1 (p70S6K) pathway and neuronal apoptosis in neonatal rats with hypoxic-ischemic encephalopathy (HIE).Methods neonatal rats HIE model were established by ligating left common carotid artery and hypoxic glass chamber, they were randomly divided into 5 groups: model group, low dose (0.8 mg/kg), medium dose (1.6 mg/kg), high dose (3.2 mg/kg) groups of cattle encephalon glycoside and ignotin, and nimodipine group (10 mg/kg), with 12 in each group, another 12 rats were set as sham operation group. After grouping, the neurobehavioral activities of rats in each group were observed and scored, the water content of brain tissue was measured, the apoptosis of cerebral cortex neurons was detected by TUNEL staining, the serum levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were detected by enzyme-linked immunosorbent assay (ELISA), the expression of SIRT1/mTOR/p70S6K pathway related proteins was detected by Western blotting. Results Compared with those in the sham operation group, the neurological deficit score, water content of brain tissue, apoptosis rate of cortical neurons, levels of serum TNF-α and IL-1β, SIRT1/mTOR/p70S6K pathway related proteins p-mTOR/mTOR and p-p70S6k/p70S6K pathway were significantly increased in the model group, the expression of SIRT1 in brain tissue was decreased (P < 0.001). Compared with those in the model group, the neurological deficit score, water content of brain tissue, apoptosis rate of cortical neurons, levels of serum TNF-α and IL-1β, SIRT1/mTOR/p70S6K pathway related proteins p-mTOR/mTOR and p-p70S6k/p70S6K pathway were significantly decreased in the low, medium and high dose groups of cattle encephalon glycoside and ignotin, and nimodipine group. The expression of SIRT1 in brain tissue was increased (P< 0.001), there was a dose-dependent effect in groups of cattle encephalon glycoside and ignotin (P < 0.001). There was no significant difference in each index between high-dose group of cattle encephalon glycoside and ignotin and nimodipine group. Conclusion Cattle encephalon glycoside and ignotin can promote the expression of SIRT1 in brain tissue of neonatal rats with HIE, reduce the phosphorylation levels of mTOR and p70S6K, inhibit inflammation, relieve brain edema and neuronal apoptosis, and repair nerve function and improve clinical symptoms.

R965.1

河南省科技攻關(guān)項(xiàng)目（LHGJ20191385）