【摘要】：目的:研究激活TRPV1在高同型半胱氨酸促內皮細胞凋亡中的作用。方法:培養(yǎng)人臍靜脈內皮細胞,用不同濃度同型半胱氨酸(0 mmol/L、0.05 mmol/L、0.1 mmol/L、0.2 mmol/L、0.5 mmol/L、1 mmol/L、2mmol/L、4mmol/L)及不同時間(6h、12h、24h、36h、48h)進行干預后CCK-8檢測細胞增殖及活性情況,選擇合適的干預濃度及干預時間;用免疫學方法檢測TRPV1是否在人臍靜脈內皮細胞上表達;Western blotting檢測各組TRPV1蛋白的表達,TUNEL染色了解各組細胞凋亡情況。結果:(1)同型半胱氨酸對人臍靜脈內皮細胞的增殖能力有明顯的抑制作用,OD值從(1.03±0.12)漸下降至(0.42±0.05),且呈明顯的劑量效應關系(P0.01);同型半胱氨酸在0.5mmol/L開始、干預24h后對人臍靜脈內皮細胞增殖有抑制作用(P0.05);(2)細胞免疫熒光檢測人臍靜脈內皮細胞上有TRPV1表達,而對照組無表達;(3)不同濃度同型半胱氨酸干預細胞后TPRV1蛋白表達及細胞凋亡有差異(P0.05);(4)辣椒素干預后TRPV1表達及細胞凋亡較單純同型半胱氨酸干預組有統(tǒng)計學差異(P0.05)。結論:高同型半胱氨酸可通過對TRPV1蛋白表達水平的下調,促進內皮細胞凋亡;激活TRPV1可抑制高同型半胱氨酸促內皮細胞凋亡。
[Abstract]:Aim: to study the effect of activating TRPV1 on the apoptosis of endothelial cells induced by homocysteine. Methods: human umbilical vein endothelial cells were cultured with different concentrations of homocysteine (0 mmol/L,0.05 mmol/L,0.1 mmol/L,0.2 mmol/L,0.5 mmol/L,1 mmol/L,2mmol/L,). 4mmol/L) and different time (6 h ~ 12 h ~ 24 h ~ 36 h ~ 48 h) were used to detect the proliferation and activity of CCK-8, and the appropriate intervention concentration and intervention time were selected. The expression of TRPV1 on human umbilical vein endothelial cells (HUVECs) was detected by immunological method. The expression of TRPV1 protein in each group was detected by TUNEL staining and apoptosis was detected by TUNEL staining. Results: (1) homocysteine significantly inhibited the proliferation of human umbilical vein endothelial cells, and the OD value decreased from (1.03 鹵0.12) to (0.42 鹵0. 05) in a dose-dependent manner (P0.01). Homocysteine inhibited the proliferation of human umbilical vein endothelial cells (P0.05); (2) at the beginning of 0.5mmol/L and 24 h after intervention (P0.05); (2). The expression of TRPV1 in human umbilical vein endothelial cells was detected by immunofluorescence, but no expression was found in the control group. (3) the expression of TPRV1 protein and cell apoptosis were different in different concentrations of homocysteine (P0.05). (4) the expression of TRPV1 and apoptosis after capsaicin intervention were significantly higher than those in homocysteine group (P0.05). Conclusion: high homocysteine can promote the apoptosis of endothelial cells by down-regulating the expression of TRPV1 protein, and activating TRPV1 can inhibit the apoptosis of endothelial cells induced by high homocysteine.
【作者單位】： 寧夏醫(yī)科大學總醫(yī)院心臟中心;寧夏醫(yī)科大學心腦血管病醫(yī)院;寧夏醫(yī)科大學;寧夏醫(yī)科大學總醫(yī)院心研所;
【基金】：寧夏醫(yī)科大學優(yōu)勢學科群建設科研項目(No:XY201501) 寧夏自然科學基金(No:NZ14113)
【分類號】：R54

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