Fang Kuai-Fa, Chen Zhu-Jun, Liu Meng, Wu Ping-Sheng, Yu Da-Zhi
Department of Cardiology, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong Province 510515, China; Department of Cardiology, Affiliated Huiyang Hospital of Southern Medical University, Huizhou, Guangdong Province 516200, China; Department of Cardiology, Huizhou No.6 People's Hospital, Huizhou, Guangdong Province 516200, China.
Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong Province 510080, China.
Asian Pac J Trop Med. 2015 Oct;8(10):864-9. doi: 10.1016/j.apjtm.2015.09.015. Epub 2015 Sep 25.
To study the mechanism and significance of pH change in the coronary artery microthrombosis of rats.
After the sodium laurate-induced model of coronary artery microthrombosis of rats was constructed, the vascular endothelial cells were separated and then cultured in the mediums with different pH values for 24 h. Enzyme linked immunosorbent assay was used to detect the content of von Willebrand factor (vWF) in the medium; while the real-time PCR and western blot assay were used to detect the expression of fibrinogen-like protein 2 (FGL2) at the mRNA and protein level. The comprehensive evaluation was performed to discuss the effect of pH change on the coronary artery microthrombosis of rats.
The expression level of vWF detected by enzyme linked immunosorbent assay was 336.67 ± 24.95, 311.33 ± 14.98, 359.67 ± 39.63, 354.67 ± 49.01 and 332.00 ± 33.42 (pg/mL) respectively; while the expression of vWF in the model group was 570.00 ± 57.94, 524.67 ± 57.94, 437.00 ± 95.38, 415.33 ± 44.38 and 444.67 ± 74.31 respectively. Being cultured under the different pH values, the relative expression level of FGL2 mRNA in the model group was 7.93 ± 0.93, 6.70 ± 0.70, 5.03 ± 0.32, 5.13 ± 0.40 and 5.57 ± 0.83 respectively.
The coronary artery microthrombosis of rats can cause the high expression and secretion of vWF. Meanwhile, FGL2 is also up-regulated in the thrombosis and such up-regulation is more significant in the condition with low pH, which indicates that the low pH condition may be one of factors that contribute to the cardiovascular diseases.
研究大鼠冠状动脉微血栓形成过程中pH变化的机制及意义。
构建月桂酸钠诱导的大鼠冠状动脉微血栓形成模型后，分离血管内皮细胞，然后在不同pH值的培养基中培养24小时。采用酶联免疫吸附测定法检测培养基中血管性血友病因子（vWF）的含量；同时采用实时荧光定量聚合酶链反应（PCR）和蛋白质免疫印迹法检测纤维蛋白原样蛋白2（FGL2）在mRNA和蛋白水平的表达。进行综合评价，探讨pH变化对大鼠冠状动脉微血栓形成的影响。
酶联免疫吸附测定法检测的vWF表达水平分别为336.67±24.95、311.33±14.98、359.67±39.63、354.67±49.01和332.00±33.42（pg/mL）；而模型组vWF的表达分别为570.00±57.94、524.67±57.94、437.00±95.38、415.33±44.38和444.67±74.31。在不同pH值条件下培养，模型组FGL2 mRNA的相对表达水平分别为7.93±0.93、6.70±0.70、5.03±0.32、5.13±0.40和5.57±0.83。
大鼠冠状动脉微血栓形成可导致vWF的高表达和分泌。同时，FGL2在血栓形成中也上调，且在低pH条件下这种上调更显著，这表明低pH条件可能是导致心血管疾病的因素之一。
