低切應(yīng)力誘導(dǎo)血管細(xì)胞活化激酶C受體1的表達(dá)及其意義
發(fā)布時(shí)間：2018-07-31 18:36
【摘要】：血管重建（remodeling）是心血管疾病共有的發(fā)病基礎(chǔ)和基本的病理過(guò)程。血流切應(yīng)力變化是影響血管重建的重要因素，與動(dòng)脈粥樣硬化的發(fā)生發(fā)展密切相關(guān)。血管內(nèi)皮細(xì)胞(endothelial cells, ECs)直接承載血流切應(yīng)力刺激，并通過(guò)細(xì)胞間相互作用影響血管平滑肌細(xì)胞（vascular smooth muscle cells, VSMCs）。研究ECs與VSMCs間的相互交流，以及切應(yīng)力下的細(xì)胞功能變化，有助于闡明血管重建的力學(xué)生物學(xué)機(jī)制。 我們通過(guò)前期蛋白質(zhì)組學(xué)的工作，得到了正常切應(yīng)力（normalshear stress, NSS,15dyn/cm2）和低切應(yīng)力（low shear stress，LowSS，5dyn/cm2）條件下體外培養(yǎng)24h的血管組織差異表達(dá)的蛋白質(zhì)譜。對(duì)比發(fā)現(xiàn)，活化激酶C受體1（receptor for activated C kinase1，RACK1）在低切應(yīng)力作用下的血管組織中呈高表達(dá)，提示RACK1可能參與了LowSS誘導(dǎo)的血管重建。 本文應(yīng)用ECs/VSMCs聯(lián)合培養(yǎng)流動(dòng)腔系統(tǒng)，獲得靜態(tài)（static）、NSS和LowSS條件下聯(lián)合培養(yǎng)大鼠ECs和VSMCs表達(dá)RACK1及Akt磷酸化水平。同時(shí)，為探討應(yīng)力調(diào)控的上述變化是通過(guò)ECs與VSMCs的直接接觸或是通過(guò)旁分泌作用，本文設(shè)計(jì)了ECs//VSMCs隔開(kāi)培養(yǎng)的體系，同樣檢測(cè)靜態(tài)、NSS和LowSS條件下ECs和VSMCs的RACK1表達(dá)狀況及Akt磷酸化水平。此外，本文還在靜態(tài)條件下，應(yīng)用RNA干擾（RNA interference, RNAi）技術(shù)特異性抑制VSMCs內(nèi)RACK1表達(dá)，探討RACK1對(duì)細(xì)胞凋亡和Akt磷酸化水平的作用。 結(jié)果顯示：①切應(yīng)力與VSMCs對(duì)ECs的RACK1表達(dá)均無(wú)顯著影響；②LowSS與細(xì)胞直接接觸共同誘導(dǎo)VSMCs表達(dá)RACK1，NSS、LowSS與旁分泌共同阻遏VSMCs表達(dá)RACK1；③NSS和LowSS都可能抑制ECs的Akt磷酸化；④旁分泌參與NSS對(duì)VSMCs的Akt磷酸化的促進(jìn)作用；⑤抑制VSMCs的RACK1表達(dá)下調(diào)細(xì)胞凋亡和caspase-3表達(dá)，上調(diào)Akt磷酸化。 上述結(jié)果表明，LowSS條件下，ECs通過(guò)直接接觸作用誘導(dǎo)VSMCs的RACK1表達(dá)；切應(yīng)力對(duì)ECs與VSMCs的Akt磷酸化水平的誘導(dǎo)作用不同，NSS和LowSS均抑制ECs的Akt磷酸化，，但NSS的抑制作用更強(qiáng)，而NSS則促進(jìn)VSMCs的Akt磷酸化；RACK1可能通過(guò)PI3K/Akt信號(hào)通路參與切應(yīng)力對(duì)細(xì)胞凋亡的調(diào)控。結(jié)果提示，ECs與VSMCs之間的相互接觸和聯(lián)系，對(duì)維持血管的正常結(jié)構(gòu)與功能的穩(wěn)定有重要作用。
[Abstract]:Revascularization of (remodeling) is the common pathogenesis and basic pathological process of cardiovascular disease. The change of blood flow shear stress is an important factor affecting vascular remodeling and is closely related to the occurrence and development of atherosclerosis. Vascular endothelial cells (endothelial cells, ECs) directly carry shear stress stimulation and influence vascular smooth muscle cells (VSMC) (vascular smooth muscle cells, VSMCs). Through intercellular interaction. The study of the interaction between ECs and VSMCs, as well as the changes of cell function under shear stress, is helpful to elucidate the mechanics and biological mechanism of vascular remodeling. We obtained the differentially expressed protein profiles of vascular tissues cultured in vitro for 24 hours under normal shear stress (normalshear stress,) 15 dyn / cm 2) and low shear stress (5 dyn / cm 2). It was found that activated kinase C receptor 1 (receptor for activated C kinase1 rack1) was highly expressed in vascular tissues under low shear stress, suggesting that RACK1 might be involved in the vascular remodeling induced by LowSS. In this paper, the expression of RACK1 and Akt phosphorylation of ECs and VSMCs in rats co-cultured with ECs/VSMCs and LowSS were obtained. At the same time, in order to investigate whether the above changes in stress regulation are through the direct contact between ECs and VSMCs or through paracrine action, the system of isolated culture of ECs//VSMCs was designed in this paper. The expression of RACK1 and the level of Akt phosphorylation in ECs and VSMCs were also detected under the static condition of NSS and LowSS. In addition, under static conditions, RNA interference (RNA interference, RNAi) technique was used to specifically inhibit the expression of RACK1 in VSMCs, and to explore the effect of RACK1 on apoptosis and Akt phosphorylation. The results showed that the RACK1 expression of ECs was not significantly affected by the shear stress of 1: 1 and VSMCs. The direct contact between VSMCs and cells induced VSMCs expression of rack1, NSS, low SS and paracrine to inhibit VSMCs expression of rack1n3NSS and LowSS, both of which might inhibit Akt phosphorylation of ECs. 4 paracrine plays an important role in the promotion of Akt phosphorylation of VSMCs by NSS. 5 inhibits the expression of RACK1 in VSMCs and down-regulates the expression of caspase-3 and apoptosis, and up-regulates the phosphorylation of Akt. The results indicated that the RACK1 expression of VSMCs was induced by direct contact between ECs and VSMCs under LowSS condition, and that shear stress induced Akt phosphorylation level of ECs and VSMCs was different. Both LowSS and ECs inhibited Akt phosphorylation of ECs, but NSS inhibited ECs phosphorylation more strongly. NSS promotes the Akt phosphorylation of VSMCs, RACK1, which may be involved in the regulation of apoptosis by shear stress through PI3K/Akt signaling pathway. The results suggest that the contact and connection between ECs and VSMCs play an important role in maintaining the normal structure and function of blood vessels.
【學(xué)位授予單位】：上海交通大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R363
本文編號(hào)：2156474
[Abstract]:Revascularization of (remodeling) is the common pathogenesis and basic pathological process of cardiovascular disease. The change of blood flow shear stress is an important factor affecting vascular remodeling and is closely related to the occurrence and development of atherosclerosis. Vascular endothelial cells (endothelial cells, ECs) directly carry shear stress stimulation and influence vascular smooth muscle cells (VSMC) (vascular smooth muscle cells, VSMCs). Through intercellular interaction. The study of the interaction between ECs and VSMCs, as well as the changes of cell function under shear stress, is helpful to elucidate the mechanics and biological mechanism of vascular remodeling. We obtained the differentially expressed protein profiles of vascular tissues cultured in vitro for 24 hours under normal shear stress (normalshear stress,) 15 dyn / cm 2) and low shear stress (5 dyn / cm 2). It was found that activated kinase C receptor 1 (receptor for activated C kinase1 rack1) was highly expressed in vascular tissues under low shear stress, suggesting that RACK1 might be involved in the vascular remodeling induced by LowSS. In this paper, the expression of RACK1 and Akt phosphorylation of ECs and VSMCs in rats co-cultured with ECs/VSMCs and LowSS were obtained. At the same time, in order to investigate whether the above changes in stress regulation are through the direct contact between ECs and VSMCs or through paracrine action, the system of isolated culture of ECs//VSMCs was designed in this paper. The expression of RACK1 and the level of Akt phosphorylation in ECs and VSMCs were also detected under the static condition of NSS and LowSS. In addition, under static conditions, RNA interference (RNA interference, RNAi) technique was used to specifically inhibit the expression of RACK1 in VSMCs, and to explore the effect of RACK1 on apoptosis and Akt phosphorylation. The results showed that the RACK1 expression of ECs was not significantly affected by the shear stress of 1: 1 and VSMCs. The direct contact between VSMCs and cells induced VSMCs expression of rack1, NSS, low SS and paracrine to inhibit VSMCs expression of rack1n3NSS and LowSS, both of which might inhibit Akt phosphorylation of ECs. 4 paracrine plays an important role in the promotion of Akt phosphorylation of VSMCs by NSS. 5 inhibits the expression of RACK1 in VSMCs and down-regulates the expression of caspase-3 and apoptosis, and up-regulates the phosphorylation of Akt. The results indicated that the RACK1 expression of VSMCs was induced by direct contact between ECs and VSMCs under LowSS condition, and that shear stress induced Akt phosphorylation level of ECs and VSMCs was different. Both LowSS and ECs inhibited Akt phosphorylation of ECs, but NSS inhibited ECs phosphorylation more strongly. NSS promotes the Akt phosphorylation of VSMCs, RACK1, which may be involved in the regulation of apoptosis by shear stress through PI3K/Akt signaling pathway. The results suggest that the contact and connection between ECs and VSMCs play an important role in maintaining the normal structure and function of blood vessels.
【學(xué)位授予單位】：上海交通大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R363
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相關(guān)期刊論文 前2條
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