本文選題：平滑肌細(xì)胞 + 細(xì)胞表型��； 參考：《中國人民解放軍軍醫(yī)進(jìn)修學(xué)院》2007年碩士論文

【摘要】： 血管移植術(shù)、血管成形術(shù)后再狹窄、高血壓病和動脈粥樣硬化形成的一個(gè)共同病理過程是血管內(nèi)膜增生，這種增生以血管中膜VSMCs向內(nèi)膜浸潤和增殖為主要特征。在VSMCs增殖之前，有一個(gè)明顯的表型改變的過程。PDGF是VSMCs分化和增殖的有絲分裂原，能夠抑制收縮蛋白SM-α-actin的表達(dá)，同時(shí)啟動VSMCs的DNA合成，促進(jìn)VSMCs由分化表型向收縮表型轉(zhuǎn)化。而HO-1是熱休克蛋白家族成員，，是體內(nèi)重要的內(nèi)源性保護(hù)蛋白之一，可以明顯抑制平滑肌細(xì)胞增殖。雷帕霉素可以對內(nèi)皮細(xì)胞的增殖和分化有抑制作用，并且可以明顯抑制VSMCs的增殖。已被廣泛應(yīng)用于藥物洗脫支架以預(yù)防血管成型術(shù)后再狹窄。為了觀察HO-1和雷帕霉素對PDGF誘導(dǎo)的大鼠VSMCs表型轉(zhuǎn)變的影響和對比HO-1與雷帕霉素作用結(jié)果的差別，設(shè)計(jì)了本試驗(yàn)研究。 目的：觀察HO-1與雷帕霉素對PDGF誘導(dǎo)的大鼠VSMCs表型轉(zhuǎn)化的作用。 方法：原代大鼠VSMCs分離培養(yǎng)，給予PDGF-BB誘導(dǎo)VSMCs表型轉(zhuǎn)化。同時(shí)給予不同劑量的血紅素氧合酶誘導(dǎo)劑氯血紅素誘導(dǎo)HO-1的表達(dá)，和雷帕霉素共培養(yǎng)24小時(shí)。免疫熒光染色后應(yīng)用激光共聚焦顯微鏡觀察各組SM-α-actin和PCNA的表達(dá)，計(jì)算各組熒光強(qiáng)度，觀察在不同處理?xiàng)l件下對VSMCs表型轉(zhuǎn)化的影響。 結(jié)果：PDGF-BB20nmol／mL作用原代VSMCs 24h后，SM-a-actin與PCNA免疫熒光染色與對照組相比較發(fā)現(xiàn)SM-a-actin表達(dá)減弱，PCNA表達(dá)增強(qiáng)。氯血紅素誘導(dǎo)HO-1后可以抑制PDGF-BB誘導(dǎo)的VSMCs表型改變，并且隨劑量的增加，抑制強(qiáng)度增加。較低劑量的雷帕霉素可以抑制PDGF誘導(dǎo)的VSMCs表型轉(zhuǎn)化。 結(jié)論：1、PDGF-BB亞型可以促進(jìn)原代培養(yǎng)的大鼠VSMCs由收縮表型向 合成表型的轉(zhuǎn)化。2、大鼠VSMCs可能在體內(nèi)即存在部分合成表型，并且在體外培養(yǎng)的過程中有自發(fā)性去分化改變。3、給予氯血紅素誘導(dǎo)HO-1表達(dá)可以對抗PDGF-BB誘導(dǎo)的大鼠VSMCs表型轉(zhuǎn)化，并呈劑量相關(guān)。4、較低劑量的雷帕霉素可以抑制大鼠VSMCs的表型轉(zhuǎn)化。
[Abstract]:A common pathological process of vascular transplantation, restenosis after angioplasty, hypertension and atherosclerosis is intimal hyperplasia, which is characterized by the infiltration and proliferation of VSMCs into the intima. Before the proliferation of VSMCs, there was an obvious phenotypic change process. PDGF was a mitogen for differentiation and proliferation of VSMCs. PDGF could inhibit the expression of SM- 偽 -actin, activate the DNA synthesis of VSMCs, and promote the transformation of VSMCs from differentiation phenotype to contractile phenotype. HO-1, a member of heat shock protein family, is one of the important endogenous protective proteins in vivo, which can inhibit the proliferation of smooth muscle cells. Rapamycin could inhibit the proliferation and differentiation of endothelial cells and inhibit the proliferation of VSMCs. It has been widely used in drug-eluting stents to prevent restenosis after angioplasty. In order to observe the effect of HO-1 and rapamycin on the phenotypic transformation of VSMCs induced by PDGF in rats and to compare the results of HO-1 and rapamycin, this experiment was designed. Aim: to observe the effects of HO-1 and rapamycin on VSMCs phenotypic transformation induced by PDGF in rats. Methods: primary rat VSMCs was isolated and cultured. PDGF-BB was used to induce VSMCs phenotypic transformation. The expression of HO-1 was induced by different doses of heme oxygenase inducer and co-cultured with rapamycin for 24 hours. After immunofluorescence staining, the expressions of SM- 偽 -actin and PCNA were observed by laser confocal microscopy, the fluorescence intensity of each group was calculated, and the effect of different treatments on the phenotypic transformation of VSMCs was observed. Results the expression of SM-a-actin and PCNA in primary VSMCs treated with 20 nmol / mL of w PDGF-BB20 nmol / mL for 24 hours was lower than that in control group. Hemin induced HO-1 could inhibit VSMCs phenotype changes induced by PDGF-BB, and the inhibition intensity increased with the increase of dosage. Lower doses of rapamycin inhibited PDGF induced phenotypic transformation of VSMCs. Conclusion the PDGF-BB subtype can promote the contractile phenotype of rat VSMCs in primary culture. The transformation of synthetic phenotype. 2. The partial synthetic phenotype of rat VSMCs may exist in vivo, and there is spontaneous dedifferentiation in the process of culture in vitro. HO-1 expression induced by hemin can antagonize the phenotypic transformation of rat VSMCs induced by PDGF-BB. Low dose rapamycin inhibited the phenotypic transformation of VSMCs in rats.
【學(xué)位授予單位】：中國人民解放軍軍醫(yī)進(jìn)修學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2007
【分類號】：R363

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相關(guān)期刊論文 前2條

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2 周俊,陸國平,戚文航;核因子κB的反義和誘騙性寡核苷酸對大鼠球囊損傷后血管狹窄和新生內(nèi)膜形成的影響[J];中國動脈硬化雜志;2003年04期

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