摘要
一、颈动-静脉分流肺动脉高压模型的建立及二氯乙酸盐对该模型的作用
【目的】探讨二氯乙酸盐(DCA)对颈动-静脉分流诱导大鼠的肺动脉高压中肺血管平滑肌细胞增殖和凋亡的影响。
【方法】将雄性SD大鼠随机分为四组:正常对照组、动物模型组、单纯结扎组、DCA+结扎联合治疗组,每组各10只。后面三组分离颈部左颈总动脉和左颈外静脉,将两者端端吻合以诱导肺动脉高压产生。从第8周开始,正常组和模型组予以等量的生理盐水喂养2周;单纯结扎组将动-静分流的血管进行结扎,再以等量的生理盐水喂养2周;DCA+结扎联合治疗组将动-静分流的血管进行结扎,再以DCA(0.75g/L,pH7.0)喂养2周,剂量为80mg/kg/d。各组分别于第10周进行开胸平均肺动脉压力(mPAP)的测量。测量后处死,取肺组织进行H-E染色、中膜厚度百分比和右心室肥厚指数测量计算、细胞增殖指标(增殖细胞核抗原PCNA)和凋亡指标(Caspase-3)的免疫组织化学染色。
【结果】10周后,平均肺动脉:与模型组相比,单纯结扎组、DCA+结扎联合治疗组压明显降低(P<0.05),但联合组与单纯结扎组相比差别无统计学意义(P>0.05)。中膜厚度百分比和右心室肥厚指数:单纯结扎组、联合组均比模型组显著降低(P<0.05),但联合组与单纯结扎组相比差别无统计学意义(P>0.05)。PCNA增殖度:单纯结扎组与模型组差异无统计学意义(P>0.05),联合组比模型组和单纯结扎组要低,差异有统计学意义(P<0.05);而Caspases-3阳性率:单纯结扎组与模型组差异无统计学意义(P>0.05),联合组比模型组和单纯结扎组要高,差异有统计学意义(P<0.05)。
【结论】DCA对早期肺血容量增加所致的动力性肺动脉高压无明显效果,但可以抑制肺动脉平滑肌细胞增殖并促进其凋亡。
二、野百合碱所致肺动脉高压模型的建立及二氯乙酸盐对该模型的作用
【目的】探讨二氯乙酸盐(DCA)对野百合碱(MCT)诱导大鼠的肺动脉高压中肺血管平滑肌增殖和凋亡的影响。
【方法】将雄性SD大鼠随机分为三组:正常组,模型组和治疗组,每组各10只。模型组及治疗组一次性皮下注射野百合碱(MCT)60mg/kg,正常组注射等量生理盐水。于注射野百合碱的第7天,治疗组予以DCA(0.75g/L,pH7.0)喂养,剂量为80mg/kg/d。正常组和模型组予以等量的生理盐水喂养。各组分别于第7、14、21、28天进行平均肺动脉压力(mPAP)的测量。28天后处死,取肺组织进行细胞增殖指标(增殖细胞核抗原PCNA)和凋亡指标(Caspase-3)的免疫组化染色和H-E染色。
【结果】平均肺动脉压测量:同时间点模型组和治疗组均高于正常组,差异均有统计学意义(P<0.05)。第7天,治疗组与模型组相比,差异无统计学意义(P>0.05)。自第14天后开始,治疗组的mPAP均低于模型组,且差异均有统计学意义(P<0.05)。中膜厚度百分比:模型组高于正常组(P<0.05),治疗组高于正常组,低于模型组(P<0.05),差异均有统计学意义。右心室肥厚指数(right ventricular hypertrophy index RVHI):模型组高于正常组(P<0.05),治疗组RVHI高于正常组,低于模型组(P<0.05),差异也均有统计学意义。PCNA增殖度:模型组高于正常组(P<0.05),治疗组高于正常组,低于模型组(P<0.05),差异均有统计学意义。Caspase-3染色阳性率:模型组低于正常组,(P<0.05),治疗组低于正常组,高于模型组(P<0.05),差异均有统计学意义。
【结论】DCA可以抑制肺动脉平滑肌细胞增殖并促进其凋亡,从而逆转肺血管重构。
PartⅠEstablishment of Pulmonary Hypertension Model by Carotid Artery to Jugular Vein Shunt, and Effect of Dichloroacetate on the Model
【Objective】To discuss the effect of Dichloroacetate (DCA) on proliferation and apoptosis of pulmonary arterial smooth muscle cells (PASMCs) in pulmonary arterial hypertension (PAH),which is induced by carotid artery to jugular vein shunt.
【Methods】It was divided into four groups randomly: Control group, model group, deligation group, and DCA+deligation group, each of which had ten rats. In latter three groups, we separated the left common carotid artery and left external jugular vein, and then induced pulmonary hypertension by end-to-end anastomosis of two vessels. 8 weeks later, the equivalent volume of isotonic Na chloride was given to the control and model groups for 2 weeks. In deligation group, we had ligated the vessel of artery to vein shunt after 8 weeks, the equivalent volume of isotonic Na chloride was given them for 2 weeks. In DCA+deligation group, we had ligated the vessel of artery to vein shunt after 8 weeks, then give DCA(0.75g/L,pH7.0) to them for 2 weeks, the dose was 80mg/kg/d. On week 10, the mean pulmonary arterial pressure (mPAP) of each group had been measured; the media tunica thickness percentage and the right ventricular hypertrophy index were measured; the lung tissue was detected with Hematoxylin-Eosin stain and immunohistochemistry stain. The latter stain was used to show the proliferation (PCNA) and apoptosis (Caspase-3) of PASMCs.
【Results】Compared to model group, the mPAP of deligation group and DCA+deligation group had decreased significantly(P < 0.05), but there was no differences between deligation group and DCA+deligation group (P>0.05). Meanwhile, Compared to model group, the media tunica thickness percentage and the right ventricular hypertrophy index (RVHI) of deligation group and DCA+deligation group reduced significantly (P<0.05), but there was no differences between deligation group and DCA+deligation group (P>0.05). PCNA of DCA+deligation group was lower than model group and deligation group (P<0.05), while there was no differences between deligation group and model group(P>0.05). Caspases-3 of DCA+deligation group was higher than model group and deligation group (P<0.05), while there was no differences between deligation group and model group(P>0.05).
【Conclusion】DCA has no significant effect on the pulmonary hypertension induced by high lung blood volume, while can restrain proliferation and promote apoptosis.
PartⅡEffect of Dichloroacetate on Monocrotaline-Induced Pulmonary Hypertension in Rat
【Objective】To discuss the effect of Dichloroacetate (DCA) on proliferation and apoptosis of pulmonary arterial smooth muscle cells (PASMCs) in Monocrotaline-induced pulmonary arterial hypertension (PAH).
【Methods】It was divided into three groups randomly: Control group, model group and DCA group, each of which had ten rats. In the model group and DCA group, the rats were subjected to single subcutaneous injection of MCT (the dose is 60mg/kg) to induce the pulmonary arterial hypertension (PAH). DCA (0.75g/L,pH7.0) was given to DCA group form day 7, the dose of which is 80mg/kg/d. Form the same time on, the equivalent volume of isotonic Na chloride was given to the other two groups. The mean pulmonary arterial pressure (mPAP) of each group had been measured on day 7, 14, 21 and 28; on day 28, the media tunica thickness percentage and the right ventricular hypertrophy index (RVHI) were measured; on day 28, the lung tissue was detected with Hematoxylin-Eosin stain and immunohistochemistry stain. The latter stain was used to show the proliferation (proliferating cell nuclear antigen, PCNA) and apoptosis (Caspase-3) of PASMCs.
【Results】The mPAP of DCA group is higher than control group and lower than model group (P<0.05). Meanwhile, The mPAP of model group is higher than control group(P<0.05). Compared with the model group, the mPAP of DCA group had decreased from day 14, which was almost half of the model group on day 28 (P<0.05). Meanwhile, Compared with the model group, the media tunica thickness percentage and the right ventricular hypertrophy index(RVHI) of DCA group reduced significantly (P<0.05). PCNA of DCA group was lower than model group (P<0.05), while Caspase-3 of DCA group was much higher than model group (P<0.05).
【Conclusion】Dichloroacetate can reverse pulmonary vascular remodeling through anti-proliferation and pro-apoptosis.
引文
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