학술논문
인간 제대정맥 내피세포에서 재조합 인간 HAPLN1 단백질의 노화 지연 효과 연구 / The Suppressive Effect of Recombinant Human HAPLN1 Protein on Senescence of Human Umbilical Vein Endothelial Cells
Document Type
Dissertation/ Thesis
Source
Subject
Language
English
Abstract
죽상동맥경화성 심혈관 질환은 심혈관계의 가장 흔한 질병으로 혈관 내막(endothelium)에 콜레스테롤 등과 같은 지방이 침착하고 내피세포의 증식이 일어나 죽종(atheroma)이 형성되며 노화에 따른 혈관의 섬유화로 혈관의 탄성이 줄어드는 질환이다. 세포 노화는 이러한 혈관 내피의 기능 장애를 유발하는 핵심 기전으로 알려져있다. 히알루론산과 프로테오글리칸 연결 단백질 1 (hyaluronan and proteoglycan link protein 1; HAPLN1)은 세포외기질(ECM)의 구성을 안정화시키는 콘드로넥신 및 ECM 성분으로, 일부 연구에서 HAPLN1은 노화와 관련이 있는 것으로 보고 되었다. 본 연구에서는 인간 제대정맥 내피세포에서 HAPLN1단백질의 내피세포 노화 억제효과를 연구하였다. 본 연구의 결과에 따르면 HAPLN1의 발현은 세포의 노화에 따라 감소되었으며, 노화 과정과 관련된 SIRT1 (NAD-dependent deacetylase sirtuin-1) 발현도 노화에 의해 감소되었다. 그러나 HAPLN1처리에 의해 SIRT1 단백질 발현이 증가하였고, SIRT1의 탈아세틸화 효소 활성은 세포 노화 바이오마커인 p53, p21 및 p16의 발현을 감소시켰다. 또한, HAPLN1은 노화 관련 β-갈락토시다아제 (SA-β-gal) 양성 세포와 IL-6, CCL2 및 IL-1β 등과 같은 노화 관련 분비 표현형(Senescence Associated Secretory Phenotype, SASP)을 감소시켰고, 혈관 내피 노화에 관여하는 IL-17 생성을 감소시켰다. HAPLN1 처리는 세포 활성산소종 생성을 감소시켜 산화 스트레스로부터 노화된 HUVECs를 보호하여 세포 생존과 기능을 향상시켰고, 노화 HUVEC의 증식, 이동 및 혈관 신생을 강화시켰다. 따라서 HAPLN1은 HUVEC에서 세포 복제와 IL-17A 유도에 의한 노화 및 산화 스트레스 유도 노화를 감소시킨다고 결론지을 수 있다. HAPLN1의 노화 방지 효과 기전에 대한 이해는 심혈관 질환(CVD) 및 죽상동맥경화증 치료에서 새로운 가능성 있는 전략으로 이어질 것이라 생각된다.
Cardiovascular diseases (CVDs) are the most common cardiovascular sys-tem disorders. Cellular senescence is a key mechanism associated with dys-function of aged vascular endothelium. Hyaluronic acid (HA) and proteogly-can (PG) link protein 1 (HAPLN1) has been known to non-covalently link HA and PGs, and forms and stabilizes HAPLN1-containing aggregates as a major component of extracellular matrix. Our previous study showed that serum lev-els of HAPLN1 decrease with aging. Here, we found that the HAPLN1 gene expression was reduced in senescent human umbilical vein endothelial cells (HUVECs). Moreover, a recombinant human HAPLN1 (rhHAPLN1) de-creased the activity of senescence-associated β-gal and inhibited the production of senescence-associated secretory phenotypes, including IL-1β, CCL2 and IL-6. rhHAPLN1 also down-regulated IL-17A levels, which is known play a key role in vascular endothelial senescence. In addition, rhHAPLN1 protected se-nescent HUVECs from oxidative stress by reducing cellular reactive oxygen species levels, thus promoting the function and survival of HUVECs and lead-ing to cellular proliferation, migration, and angiogenesis. We also found that rhHAPLN1 not only increases the sirtuin 1 (SIRT1) levels, but also reduces the cellular senescence markers levels, such as p53, p21, and p16. Taken together, our data indicate that rhHAPLN1 delays or inhibits the endothelial senescence induced by various aging factors, such as DNA replication, IL-17A, and oxida-tive stress, thus suggesting that rhHAPLN1 may be a promising therapeutic for CVD and atherosclerosis.
Cardiovascular diseases (CVDs) are the most common cardiovascular sys-tem disorders. Cellular senescence is a key mechanism associated with dys-function of aged vascular endothelium. Hyaluronic acid (HA) and proteogly-can (PG) link protein 1 (HAPLN1) has been known to non-covalently link HA and PGs, and forms and stabilizes HAPLN1-containing aggregates as a major component of extracellular matrix. Our previous study showed that serum lev-els of HAPLN1 decrease with aging. Here, we found that the HAPLN1 gene expression was reduced in senescent human umbilical vein endothelial cells (HUVECs). Moreover, a recombinant human HAPLN1 (rhHAPLN1) de-creased the activity of senescence-associated β-gal and inhibited the production of senescence-associated secretory phenotypes, including IL-1β, CCL2 and IL-6. rhHAPLN1 also down-regulated IL-17A levels, which is known play a key role in vascular endothelial senescence. In addition, rhHAPLN1 protected se-nescent HUVECs from oxidative stress by reducing cellular reactive oxygen species levels, thus promoting the function and survival of HUVECs and lead-ing to cellular proliferation, migration, and angiogenesis. We also found that rhHAPLN1 not only increases the sirtuin 1 (SIRT1) levels, but also reduces the cellular senescence markers levels, such as p53, p21, and p16. Taken together, our data indicate that rhHAPLN1 delays or inhibits the endothelial senescence induced by various aging factors, such as DNA replication, IL-17A, and oxida-tive stress, thus suggesting that rhHAPLN1 may be a promising therapeutic for CVD and atherosclerosis.