动脉内膜损伤后血管平滑肌细胞表型转化及MKP-1表达的变化

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目的:观察动脉内膜损伤后血管平滑肌细胞(vascu lar smooth musc le cells,VSMC)表型转化和丝裂原激活蛋白激酶磷酸酶-1(m itogen-activated prote in k inase phosphatase-1,MKP-1)表达的动态变化。方法:分别用HE染色、免疫组化和逆转录-聚合酶链(RT-PCR)方法检测假损伤组(S组)和损伤后不同时间点血管形态学改变及血管壁中增殖细胞核抗原(PCNA)、平滑肌α肌动蛋白(SMα-actin)和MKP-1 mRNA及蛋白表达的变化。结果:①损伤后1 d中膜腔侧、3 d管腔内表面可见增殖的VSMC,5~7 d新生内膜(neointim a,NI)形成并逐渐增厚,14~35 d NI进行性增厚;各组中膜均有增殖的VSMC向腔面集聚。②S组中膜VSMC及内皮细胞PCNA为阴性;中膜于损伤后1~14d,NI于5~14 d PCNA阳性细胞率逐渐增多,14 d达高峰,28 d后开始逐渐减少,但NI阳性率多于中膜。③S组中膜SMα-actin表达为阳性,内皮为阴性;中膜阳性面积于损伤后1 d开始减少,3 d最为明显,5 d后开始逐渐增加,NI阳性表达弱于中膜。④S组中膜MKP-1呈弱阳性或阳性表达,损伤后1d即开始下降,5~7 d达最低,14 d稍有回升,至35 d仍未回到假损伤组水平;NI阳性表达弱于中膜。MKP-1表达变化与PCNA表达变化呈负相关。结论:VSMC增殖能力与其表型转化密切相关,MKP-1参与了损伤后VSMC表型转化的调节。 OBJECTIVE: To observe the phenotypic changes of vascu-lar smooth muscle cells (VSMCs) and the expression of mitogen-activated prote in kinases phosphatase-1 (MKP- 1) the dynamic changes of expression. Methods: The morphological changes of blood vessels and the expression of proliferating cell nuclear antigen (PCNA) in blood vessel wall were detected by HE staining, immunohistochemistry and reverse transcriptase-polymerase chain reaction (RT-PCR) ), The changes of SMα-actin and MKP-1 mRNA and protein expression. RESULTS: On day 1 after injury, proliferating VSMCs were seen on the inner side of the lumen and on the 3rd day. Neointila (NI) formed on the 5th to 7th day and gradually increased. On the 14th to 35th day, Thick; all groups of proliferating VSMC in the media to the cavity surface accumulation. (2) The mesangial VSMC and endothelial cell PCNA were negative in group S; on day 1 to 14 after injury, the rate of PCNA positive cells gradually increased from 5 to 14 days in NI and peaked on day 14, and gradually decreased after 28 days, but NI positive rate More than the membrane. ③ The expression of SMα-actin was positive in S group and negative in endothelium. The area of ​​medial membrane was decreased on the 1st day after injury, the most obvious was at 3 days, and gradually increased after 5 days. The positive expression of NI was weaker than that in the media. ④ The expression of MKP-1 in S group was weakly positive or positive, and began to decline on the 1st day after injury. It reached the lowest level on the 5th ~ 7th day and rose slightly on the 14th day, but still not returned to the level of the sham injury group on the 35th day. The NI positive expression was weak In the membrane. The change of MKP-1 expression was negatively correlated with the change of PCNA expression. CONCLUSION: VSMC proliferative ability is closely related to phenotypic transformation. MKP-1 is involved in the regulation of VSMC phenotype after injury.
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