诱导血红素氧合酶-1表达及雷帕霉素抑制PDGF诱导的大鼠平滑肌细胞表型转化的试验研究

发布时间：2018-05-21 01:34

  本文选题：平滑肌细胞 + 细胞表型　； 参考：《中国人民解放军军医进修学院》2007年硕士论文

【摘要】： 血管移植术、血管成形术后再狭窄、高血压病和动脉粥样硬化形成的一个共同病理过程是血管内膜增生，这种增生以血管中膜VSMCs向内膜浸润和增殖为主要特征。在VSMCs增殖之前，有一个明显的表型改变的过程。PDGF是VSMCs分化和增殖的有丝分裂原，能够抑制收缩蛋白SM-α-actin的表达，同时启动VSMCs的DNA合成，促进VSMCs由分化表型向收缩表型转化。而HO-1是热休克蛋白家族成员，，是体内重要的内源性保护蛋白之一，可以明显抑制平滑肌细胞增殖。雷帕霉素可以对内皮细胞的增殖和分化有抑制作用，并且可以明显抑制VSMCs的增殖。已被广泛应用于药物洗脱支架以预防血管成型术后再狭窄。为了观察HO-1和雷帕霉素对PDGF诱导的大鼠VSMCs表型转变的影响和对比HO-1与雷帕霉素作用结果的差别，设计了本试验研究。 目的：观察HO-1与雷帕霉素对PDGF诱导的大鼠VSMCs表型转化的作用。 方法：原代大鼠VSMCs分离培养，给予PDGF-BB诱导VSMCs表型转化。同时给予不同剂量的血红素氧合酶诱导剂氯血红素诱导HO-1的表达，和雷帕霉素共培养24小时。免疫荧光染色后应用激光共聚焦显微镜观察各组SM-α-actin和PCNA的表达，计算各组荧光强度，观察在不同处理条件下对VSMCs表型转化的影响。 结果：PDGF-BB20nmol／mL作用原代VSMCs 24h后，SM-a-actin与PCNA免疫荧光染色与对照组相比较发现SM-a-actin表达减弱，PCNA表达增强。氯血红素诱导HO-1后可以抑制PDGF-BB诱导的VSMCs表型改变，并且随剂量的增加，抑制强度增加。较低剂量的雷帕霉素可以抑制PDGF诱导的VSMCs表型转化。 结论：1、PDGF-BB亚型可以促进原代培养的大鼠VSMCs由收缩表型向 合成表型的转化。2、大鼠VSMCs可能在体内即存在部分合成表型，并且在体外培养的过程中有自发性去分化改变。3、给予氯血红素诱导HO-1表达可以对抗PDGF-BB诱导的大鼠VSMCs表型转化，并呈剂量相关。4、较低剂量的雷帕霉素可以抑制大鼠VSMCs的表型转化。
[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- 伪 -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- 伪 -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.
【学位授予单位】：中国人民解放军军医进修学院
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R363

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