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)至實驗35d。3、各組大鼠在實驗35d后,測定平均肺動脈壓力(mPAP),計算右心肥厚指數(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。5、采用RT-PCR測定各組大鼠肺組織中Akt mRNA相對表達水平。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)計學意義(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)計學意義(p0.05)。3、RT-PCR結(jié)果顯示模型組Akt mRNA的表達水平較對照組有增高,差異有統(tǒng)計學意義(p0.05),Western blot結(jié)果顯示模型組肺組織Akt和p-Akt蛋白表達水平較對照組亦有增高,差異有統(tǒng)計學意義(p0.05)。采用胱胺二鹽酸干預(yù)后,干預(yù)組Akt mRNA的表達水平、Akt和p-Akt蛋白表達量較模型組均有不同程度的下降,差異有統(tǒng)計學意義(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.
【學位授予單位】：西南醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：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.
【學位授予單位】：西南醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R544.1
【參考文獻】
相關(guān)期刊論文 前10條
1 王朝輝;安永;;干細胞治療肺動脈高壓的進展[J];中國胸心血管外科臨床雜志;2016年03期
2 劉桂園;宋揚;魯曉晴;趙倩倩;;肌醇六磷酸對PI3K/Akt通路中Akt蛋白和基因表達影響[J];青島大學醫(yī)學院學報;2015年05期
3 劉迎春;孫曉紅;;TG2在宮頸癌及卵巢癌中的作用[J];中國腫瘤生物治療雜志;2015年02期
4 沈凱;袁國裕;陳士良;;血管緊張素Ⅱ通過下調(diào)血管外膜過氧化氫酶促進血管重塑的研究[J];浙江醫(yī)學;2014年11期
5 胡煜;李剛;賈鵬;孫玉琴;付杰;盧翠俠;劉斌;;ERK1/2和PI3K/PKB信號通路在調(diào)節(jié)大鼠肺動脈平滑肌細胞外基質(zhì)基因表達中的作用[J];臨床兒科雜志;2013年12期
6 鄭曉宇;覃家錦;李波;何巍;馮旭;;大鼠肺動脈高壓動物模型的建立及磷酸二酯酶1C的表達[J];廣西醫(yī)學;2013年04期
7 謝婧雯;劉曉燕;;Akt信號通路在紫紺型先天性心臟病患兒心肌慢性缺氧適應(yīng)中的作用[J];臨床心血管病雜志;2012年05期
8 羅鋒;向小勇;趙興吉;;大鼠左全肺切除致右心室重塑的研究[J];第三軍醫(yī)大學學報;2010年15期
9 張清友;杜軍保;;肺動脈高壓的治療現(xiàn)狀與進展[J];臨床兒科雜志;2010年07期
10 劉斌;王獻民;魏麗;周同甫;劉瀚e，
本文編號：2407719
本文鏈接：http://sikaile.net/yixuelunwen/xxg/2407719.html
最近更新
教材專著
