【摘要】：目的既往研究結(jié)果顯示維生素D缺乏與心室重構(gòu)的發(fā)生密切相關(guān),但具體機(jī)制尚未明確。氧化應(yīng)激增加在心室重構(gòu)發(fā)生與進(jìn)展過程中發(fā)揮重要的作用,文中擬探討維生素D缺乏對(duì)小鼠心肌氧化應(yīng)激水平的影響及可能相關(guān)的信號(hào)通路。方法實(shí)驗(yàn)采用3周齡的C57小鼠,隨機(jī)數(shù)字表法分成3組:維生素D缺乏組(維生素D缺乏飼料喂養(yǎng)10周),維生素D充足組(維生素D充足飼料喂養(yǎng)10周)及維生素D缺乏后補(bǔ)充組(維生素D缺乏飼料喂養(yǎng)10周后,轉(zhuǎn)為維生素D充足飼料喂養(yǎng)+骨化三醇治療10周)。建模成功后,檢測小鼠血液中25羥維生素D3水平及相關(guān)生化指標(biāo),心臟超聲檢查小鼠心臟腔室內(nèi)徑,8-OHDG染色測定小鼠心肌陽性細(xì)胞數(shù),分別提取心肌線粒體蛋白、細(xì)胞核蛋白及總蛋白,Western blot檢查TXNIP、ASK-1、P-ASK-1、細(xì)胞色素C等蛋白表達(dá)量。結(jié)果心臟超聲結(jié)果顯示維生素D充足組小鼠與維生素D缺乏組小鼠相比較,心肌的左室舒張末期直徑及左室質(zhì)量指數(shù)均顯著增高[(3.820±0.125)mm vs(3.748±0.092)mm,(119.30±8.54)vs(97.60±3.65),P0.05)。在對(duì)小鼠心肌切片進(jìn)行8-OHDG染色后發(fā)現(xiàn),維生素D充足組小鼠心肌中8-OHDG染色陽性的心肌細(xì)胞數(shù)(65.4±2.3)顯著高于維生素D缺乏組(21.8±1.6),而補(bǔ)充骨化三醇后,可使8-OHDG染色陽性的心肌細(xì)胞數(shù)明顯回落(36.4±1.5)。Western blot檢查結(jié)果提示,維生素D充足組小鼠心肌中TXNIP蛋白的表達(dá)顯著上調(diào),并伴隨有P-ASK1/ASK-1的比值增加,Cyt C的釋放及Cleaved caspase3的增加。結(jié)論維生素D缺乏可以引起心肌的氧化應(yīng)激損傷,而損傷發(fā)生的機(jī)制可能與TXNIP蛋白表達(dá)的上調(diào)及ASK-1相關(guān)的細(xì)胞凋亡通路的激活有關(guān)。
[Abstract]:Objective previous studies have shown that vitamin D deficiency is closely related to ventricular remodeling, but the mechanism is not clear. The increase of oxidative stress plays an important role in the development and progression of ventricular remodeling. This paper aims to explore the effects of vitamin D deficiency on the level of oxidative stress in myocardium of mice and the related signaling pathways. Methods three week-old C57 mice were randomly divided into three groups: vitamin D deficiency group (fed with vitamin D deficiency for10 weeks). Vitamin D group (vitamin D sufficient feed feeding for 10 weeks) and vitamin D deficiency supplementation group (vitamin D deficiency feed feeding for 10 weeks, fed with vitamin D sufficient feed for 10 weeks to feed ossifying triol treatment for 10 weeks). After the establishment of the model, the levels of 25 hydroxyvitamin D _ 3 in blood and related biochemical indexes were detected, the heart cavity diameter was examined by echocardiography, the number of positive cells in myocardium was determined by 8-OHDG staining, and the myocardial mitochondrial protein was extracted, respectively. Nuclear protein and total protein, Western blot were used to detect the expression of TXNIP,ASK-1,P-ASK-1, cytochrome C and other proteins. Results the results of echocardiography showed that the left ventricular end-diastolic diameter and left ventricular mass index in the vitamin D group were significantly higher than those in the vitamin D deficiency group [(3.820 鹵0.125) mm vs (3.748 鹵0.092) mm,]. (119.30 鹵8.54) vs (97.60 鹵3.65, P0.05). After 8-OHDG staining in myocardial sections of mice, it was found that the number of myocardial cells positive for 8-OHDG staining in sufficient vitamin D group (65.4 鹵2.3) was significantly higher than that in vitamin D deficient group (21.8 鹵1.6). However, the number of myocardial cells positive for 8-OHDG staining was significantly decreased after supplementation of calcitriol (36.4 鹵1.5). Western blot). The results showed that the expression of TXNIP protein was significantly up-regulated in the myocardium of mice with sufficient vitamin D. The ratio of P-ASK1/ASK-1 increased the release of, Cyt C and the increase of Cleaved caspase3. Conclusion Vitamin D deficiency can induce oxidative stress injury in myocardium, and the mechanism of injury may be related to the up-regulation of TXNIP protein expression and the activation of apoptosis pathway associated with ASK-1.
【作者單位】： 南京軍區(qū)南京總醫(yī)院心內(nèi)科;
【基金】：南京軍區(qū)總醫(yī)院科研基金(2013048)
【分類號(hào)】：R591.44

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