Expression and Role of KIP1 Ubiquitylation-promoting Complex 1 in Neointimal after Carotid Arteries Balloon Injury in Rats

doi:10.3969/j.issn.1674-5817.2016.03.001

Abstract

Abstract: Objective To study the expression of Kip1 ubiquitylation-promoting complex 1(KPC1) protein after carotid arteries balloon injury in SD rats and to explore its biological function in neointimal formation process. Methods The adult male SD rats (450-500 g) were randomly divided into sham group and carotid artery injury model group. The left damaged carotid arteries were considered as the experimental group after modelling, with the left normal carotid arteries of sham group being as control group. After 1, 3, 7 ,14 and 28 days of balloon injury, SD rats of the experimental group were sacrificed and harvested for histomorphology and molecular markers assay. Hematoxylin-eosin (HE)staining was used to assess the changes of neointimal formation in the carotid artery while Western blot and qRT-PCR were used to evaluate the expression of KPC1. What’s more, Western blot was also used to assess the levels of proliferating cell nuclear antigen (PCNA). The proliferation model was established by culturing vascular smooth muscle cells (VSMCs) in vitro, then detected the KPC1 protein, KPC1 mRNA expression by Western blot and qRT-PCR. Results Western blot and qRT-PCR showed that the KPC1 expression levels of the experimental group were significantly lower than that of control group (P<0.05), while the PCNA expression was significantly higher than that of control group (P<0.05). Compared to 1 day and 3 days after carotid arteries injury, for VSMC hyperplasia, neointimal generation and luminal stenosis were observed at 7 days by HE staining. With VSMCs hyperplasia, neointimal and luminal stenosis were getting more and more serious, the thickness of intimal exceeded medial at 14 days. At 28 days, the irregular intimal hyperplasia was continued and the degree of vascular stenosis was more than 50%. After stimulation of VSMCs proliferation by platelet-derived growth factor BB (PDGF-BB) in vitro, then Western blot and qRT-PCR data showed a gradual decreasing trend of KPC1 at protein and mRNA levels. Conclusion Inhibiting KPC1 expression can significantly decrease intima hyperplasia in this model, and its mechanism may be related to the promotion of VSMCs proliferation.

Key words: Kip1 ubiquitylation-promoting complex 1(KPC1), Neointima, Vascular smooth muscle cells (VSMCs), Hyperplasia

CLC Number:

Q95-33

YANG Bo, LIN Xin-duo, WANG Xing-ren, TANG Jun-ming, YANG Jian-ye, ZHANG Lei, CAO Teng, JIANG Feng-bo, WANG Jia-ning. Expression and Role of KIP1 Ubiquitylation-promoting Complex 1 in Neointimal after Carotid Arteries Balloon Injury in Rats[J]. Laboratory Animal and Comparative Medicine, 2016, 36(3): 161-167.

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