TG2抑制劑在減輕大鼠肺動脈高壓肺血管重構(gòu)中的作用機(jī)制
發(fā)布時間：2019-01-12 11:24
【摘要】：目的:探討TG2抑制劑在減輕大鼠肺動脈高壓肺血管重構(gòu)中的作用機(jī)制。方法:1、采用左肺切除加野百合堿注射建立大鼠肺動脈高壓模型,利用TG2抑制劑─胱胺二鹽酸,抑制肺動脈高壓肺血管重構(gòu)。2、將30只健康雄性SD大鼠隨機(jī)分為3組(每組10只),即空白對照組:不做任何處理,同其他兩組大鼠相同環(huán)境下飼養(yǎng);模型組:左肺切除術(shù)7d后野百合堿(MCT,60mg/kg)項(xiàng)背部皮下注射;干預(yù)組:在模型組基礎(chǔ)上,于左肺切除術(shù)5d后,每日腹腔注射胱胺二鹽酸(112mg/kg)持續(xù)至實(shí)驗(yàn)35d。3、各組大鼠在實(shí)驗(yàn)35d后,測定平均肺動脈壓力(mPAP),計(jì)算右心肥厚指數(shù)(RVHI),即右心室游離壁(RV)與左心室+室間隔(LV+S)之比,即RV/(LV+S)。4、將各組大鼠肺組織做HE染色、肺彈力纖維染色,觀察肺血管及肺組織的病理改變,計(jì)算肺小動脈中膜厚度百分比(WT%)、肺小動脈管壁面積與血管總面積比值(WA%)、肺小動脈新生內(nèi)膜增殖度等指標(biāo)。5、采用RT-PCR測定各組大鼠肺組織中Akt mRNA相對表達(dá)水平。6、采用蛋白質(zhì)免疫印跡法(Western blot)測定各組大鼠Akt和p-Akt蛋白表達(dá)水平。結(jié)果:1、與對照組比較,采用左肺切除加野百合堿注射的模型組大鼠形成了嚴(yán)重的肺動脈高壓及右心室肥大,并有新生內(nèi)膜形成,mPAP、右心肥厚指數(shù)百分比(RVHI%)、肺小動脈中膜厚度百分比(WT%)、肺小動脈管壁面積與血管總面積比值(WA%)及新生內(nèi)膜增殖度百分比均明顯高于對照組,差異均有統(tǒng)計(jì)學(xué)意義(p0.05)。2、采用胱胺二鹽酸干預(yù)后,干預(yù)組mPAP、右心肥厚指數(shù)百分比(RVHI%)、肺小動脈中膜厚度百分比(WT%)、肺小動脈管壁面積與血管總面積比值(WA%)及新生內(nèi)膜增殖度百分比較模型組均有不同程度的降低,差異均有統(tǒng)計(jì)學(xué)意義(p0.05)。3、RT-PCR結(jié)果顯示模型組Akt mRNA的表達(dá)水平較對照組有增高,差異有統(tǒng)計(jì)學(xué)意義(p0.05),Western blot結(jié)果顯示模型組肺組織Akt和p-Akt蛋白表達(dá)水平較對照組亦有增高,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。采用胱胺二鹽酸干預(yù)后,干預(yù)組Akt mRNA的表達(dá)水平、Akt和p-Akt蛋白表達(dá)量較模型組均有不同程度的下降,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。結(jié)論:1、左肺切除+野百合堿注射成功建立大鼠肺動脈高壓模型,并形成了新生內(nèi)膜及典型病理學(xué)特征─肺血管重構(gòu)。2、TG2抑制劑在一定程度上可以抑制肺動脈高壓的形成及阻止肺血管重構(gòu);3、PI3K/Akt信號通路可能在TG2抑制劑抑制肺動脈高壓大鼠肺血管重構(gòu)中發(fā)揮重要作用。
[Abstract]:Objective: to investigate the mechanism of TG2 inhibitor on pulmonary vascular remodeling in rats with pulmonary hypertension. Methods: 1. Pulmonary hypertension model in rats was established by left pneumonectomy and monocrotaline injection. TG2 inhibitor-cysteamine dihydrochloric acid was used to inhibit pulmonary vascular remodeling. Thirty healthy male SD rats were randomly divided into 3 groups (10 rats in each group). The model group was treated with subcutaneous injection of monocrotaline (MCT,60mg/kg) on the back 7 days after left pneumonectomy. Intervention group: on the basis of the model group, 5 days after left pneumonectomy, daily intraperitoneal injection of cysteamine dihydrochloric acid (112mg/kg) continued until 35d.3.The mean pulmonary artery pressure (mPAP),) was measured after 35 days in each group. The ratio of right ventricular free wall (RV) to left ventricular septal (LV S) (RV/ (LV S). 4) was calculated by calculating the right ventricular hypertrophy index (RVHI),). The lung tissues in each group were stained with HE and the lung elastic fibers were stained. The pathological changes of pulmonary vessels and tissues were observed, the percentage of pulmonary arterioles medial thickness (WT%), the ratio of pulmonary arteriole wall area to total area (WA%), the proliferative degree of neointima of pulmonary arterioles were calculated. RT-PCR was used to detect the relative expression of Akt mRNA in the lung tissue of rats in each group. 6. The expression of Akt and p-Akt protein was detected by Western blot (Western blot). Results: 1. Compared with the control group, the rats in the model group treated with left lung resection and monocrotaline injection developed severe pulmonary hypertension and right ventricular hypertrophy, and formed neointima, mPAP, right cardiac hypertrophy index (RVHI%). The percentage of pulmonary arterioles media thickness (WT%), the ratio of pulmonary arteriole wall area to total area (WA%) and the percentage of neointimal proliferation were significantly higher than those of the control group (p0.05). After the intervention of cysteamine dihydrochloric acid, the right cardiac hypertrophy index (RVHI%) and pulmonary arteriole media thickness (WT%) were measured in the intervention group. The ratio of pulmonary arteriole wall area to total vascular area (WA%) and the percentage of neointimal proliferation in the model group were significantly lower than those in the model group (p0.05). The results of RT-PCR showed that the expression of Akt mRNA in the model group was higher than that in the control group, and the difference was statistically significant (p0.05), Western blot result showed that the expression of Akt and p-Akt protein in the lung tissue of the model group was also higher than that in the control group. The difference was statistically significant (p0.05). After the intervention of cysteamine dihydrochloric acid, the expression level of Akt mRNA, Akt and p-Akt protein in the intervention group were lower than those in the model group, and the difference was statistically significant (p0.05). Conclusion: 1. The pulmonary hypertension model was successfully established by injection of monocrotaline into the left lung, and the neointima and typical pathological features, pulmonary vascular remodeling, were formed. To some extent, TG2 inhibitor can inhibit the formation of pulmonary hypertension and prevent pulmonary vascular remodeling. The PI3K / Akt signaling pathway may play an important role in the inhibition of pulmonary vascular remodeling by TG2 inhibitors in rats with pulmonary hypertension.
【學(xué)位授予單位】：西南醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R544.1
[Abstract]:Objective: to investigate the mechanism of TG2 inhibitor on pulmonary vascular remodeling in rats with pulmonary hypertension. Methods: 1. Pulmonary hypertension model in rats was established by left pneumonectomy and monocrotaline injection. TG2 inhibitor-cysteamine dihydrochloric acid was used to inhibit pulmonary vascular remodeling. Thirty healthy male SD rats were randomly divided into 3 groups (10 rats in each group). The model group was treated with subcutaneous injection of monocrotaline (MCT,60mg/kg) on the back 7 days after left pneumonectomy. Intervention group: on the basis of the model group, 5 days after left pneumonectomy, daily intraperitoneal injection of cysteamine dihydrochloric acid (112mg/kg) continued until 35d.3.The mean pulmonary artery pressure (mPAP),) was measured after 35 days in each group. The ratio of right ventricular free wall (RV) to left ventricular septal (LV S) (RV/ (LV S). 4) was calculated by calculating the right ventricular hypertrophy index (RVHI),). The lung tissues in each group were stained with HE and the lung elastic fibers were stained. The pathological changes of pulmonary vessels and tissues were observed, the percentage of pulmonary arterioles medial thickness (WT%), the ratio of pulmonary arteriole wall area to total area (WA%), the proliferative degree of neointima of pulmonary arterioles were calculated. RT-PCR was used to detect the relative expression of Akt mRNA in the lung tissue of rats in each group. 6. The expression of Akt and p-Akt protein was detected by Western blot (Western blot). Results: 1. Compared with the control group, the rats in the model group treated with left lung resection and monocrotaline injection developed severe pulmonary hypertension and right ventricular hypertrophy, and formed neointima, mPAP, right cardiac hypertrophy index (RVHI%). The percentage of pulmonary arterioles media thickness (WT%), the ratio of pulmonary arteriole wall area to total area (WA%) and the percentage of neointimal proliferation were significantly higher than those of the control group (p0.05). After the intervention of cysteamine dihydrochloric acid, the right cardiac hypertrophy index (RVHI%) and pulmonary arteriole media thickness (WT%) were measured in the intervention group. The ratio of pulmonary arteriole wall area to total vascular area (WA%) and the percentage of neointimal proliferation in the model group were significantly lower than those in the model group (p0.05). The results of RT-PCR showed that the expression of Akt mRNA in the model group was higher than that in the control group, and the difference was statistically significant (p0.05), Western blot result showed that the expression of Akt and p-Akt protein in the lung tissue of the model group was also higher than that in the control group. The difference was statistically significant (p0.05). After the intervention of cysteamine dihydrochloric acid, the expression level of Akt mRNA, Akt and p-Akt protein in the intervention group were lower than those in the model group, and the difference was statistically significant (p0.05). Conclusion: 1. The pulmonary hypertension model was successfully established by injection of monocrotaline into the left lung, and the neointima and typical pathological features, pulmonary vascular remodeling, were formed. To some extent, TG2 inhibitor can inhibit the formation of pulmonary hypertension and prevent pulmonary vascular remodeling. The PI3K / Akt signaling pathway may play an important role in the inhibition of pulmonary vascular remodeling by TG2 inhibitors in rats with pulmonary hypertension.
【學(xué)位授予單位】：西南醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R544.1
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