降低脂筏内integrin β1减轻HPS大鼠血清诱导PMVECs增殖和迁移

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (10): 1417-1422.

降低脂筏内integrin β1减轻HPS大鼠血清诱导PMVECs增殖和迁移

高静¹, 周家奇^2*

1.浙江大学医学院附属儿童医院麻醉科,浙江杭州 310000;
2.浙江大学医学院第一附属医院重症医学科,浙江杭州 310000

收稿日期:2021-04-02 修回日期:2021-07-07 发布日期:2021-09-29
通讯作者: *695783334@qq.com
基金资助:
国家自然科学基金(81901937)

Decreased integrin β1 in lipid raft inhibits proliferation and migration of HPS rat serum-induced PMVECs

GAO Jing¹, ZHOU Jia-qi^2*

1. Department of Anesthesiology, the Children Hospital, Zhejiang University School of Medicine, Hangzhou 310000;
2. Department of ICU, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310000, China

Received:2021-04-02 Revised:2021-07-07 Published:2021-09-29
Contact: *695783334@qq.com

摘要/Abstract

摘要： 目的探讨脂筏内整合素β1(integrin β1)在肝肺综合征(HPS)大鼠血清诱导肺微血管内皮细胞(PMVECs)增殖和迁移中的作用机制。方法将PMVECs分为4组:对照组:健康大鼠血清刺激细胞;HPS组:HPS大鼠血清刺激细胞;MβCD组:事先用0.01 mol/L MβCD预处理细胞1 h,再用HPS大鼠血清刺激细胞;si-integrin β1组:利用siRNA技术构建integrin β1低表达细胞,再用HPS大鼠血清刺激细胞。采用Western blot分别检测integrin β1、FAK和pY397FAK在PMVECs脂筏内外的表达情况;采用CCK-8法检测细胞增殖改变;细胞划痕实验检测细胞迁移改变;采用ELISA检测血管内皮生长因子(VEGF)的分泌情况。结果与对照组比较,HPS组脂筏内integrin β1表达明显增加(P＜0.05),黏着斑激酶(FAK)活化水平显著增加,细胞增殖、迁移和VEGF分泌显著增加(P＜0.05);与HPS组比较,HPS+ MβCD组在给予MβCD扰乱脂筏完整结构后,脂筏内integrin β1表达受到明显抑制(P＜0.05);与HPS组比较,在给予MβCD或小干扰RNA抑制integrin β1表达后,FAK活化水平受到显著抑制,细胞增殖、迁移和VEGF分泌显著降低(P＜0.05)。结论 HPS大鼠血清通过促进PMVECs 脂筏内integrin β1表达上调从而激活integrin β1/FAK通路进而促进细胞增殖、迁移和VEGF分泌,介导了HPS肺血管重塑效应。

关键词: 肝肺综合征(HPS), 整合素β1, 肺微血管内皮细胞(PMVECs), 血管内皮生长因子(VEGF)

Abstract: Objective To investigate the effects of integrin β1 within lipid raft on PMVECs. Methods PMVECs were divided into four groups: control group; HPS group: HPS; group and MβCD group: the cells were pretreated with 0.01 mol/L MβCD for 1 hour, and then cultured with HPS rat serum; si-integrin β1 group: construction of integrin β1 low expression cells using siRNA technology, and then cultured with HPS rat serum. The expression of integrin β1,FAK and pY397FAK in lipid rafts of PMVECs in each group was detected by Western blot; cell pro- liferation, migration were observed by CCK-8 and wound healing assay; the secretion of angiogenic factors (VEGF) was detected by ELISA. Results Compared with the control group, the expression of integrin β1 and the level of activated FAK in the lipid rafts of HPS group were higher than that of the control group(P＜0.05), PMVECs proliferation, migration and the level of secreted VEGF were significantly increased(P＜0.05). Compared with the HPS group, the expression of integrin β1 and the level of activated FAK in the lipid rafts were significantly reduced in the MβCD group or in siRNA integrin β1 group(P＜0.05), PMVECs proliferation, migration and secretion of VEGF were also significantly reduced(P＜0.05). Conclusions The up-regulation of integrin β1 within lipid raft activates integrin β1/FAK pathway thereby promotes cell proliferation, migration and secretion of VEGF, which may mediate pulmonary vascular remodeling of HPS.

Key words: hepatopulmonary syndrome(HPS), integrin β1, pulmonary microvascular endothelial cells (PMVECs), vascular endothelial growth factor (VEGF)

中图分类号:

R73.3

高静, 周家奇. 降低脂筏内integrin β1减轻HPS大鼠血清诱导PMVECs增殖和迁移[J]. 基础医学与临床, 2021, 41(10): 1417-1422.

GAO Jing, ZHOU Jia-qi. Decreased integrin β1 in lipid raft inhibits proliferation and migration of HPS rat serum-induced PMVECs[J]. Basic & Clinical Medicine, 2021, 41(10): 1417-1422.

参考文献

[1]Lv Y, Fan D. Hepatopulmonary Syndrome[J]. Dig Dis Sci, 2015, 60: 1914-1923.
[2]Ondhia M, Mohamed S, Agarwal S. Persistent hypoxae-mia following pneumonia unmasking hepatopulmonary syndrome[J]. BMJ Case Rep, 2020,13:e235626. doi:10.1136/bcr-2020-235626.
[3]Pedeli X, Varin C. Pairwise likelihood estimation of latent autoregressive count models[J]. Stat Methods Med Res,2020, 29:3278-3293.
[4]Shen CC, Chen B, Gu JT, et al. AMD3100 treatment attenuates pulmonary angiogenesis by reducing the c-kit (+) cells and its pro-angiogenic activity in CBDL rat lungs[J]. Biochim Biophys Acta Mol Basis Dis,2018, 1864:676-684.
[5]Raevens S, Geerts A, Devisscher L, et al. Recent advances in the approach to hepatopulmonary syndrome and portopulmonary hypertension[J]. Acta Gastroenterol Belg,2021, 84:95-99.
[6]Rose SCP, Cunha DV, Medeiros SBC, et al. Correlation between hepatopulmonary syndrome and oxygen saturation pulse oximetry in cirrhotic patients[J]. Rev Assoc Med Bras,2020, 66:1577-1582.
[7]Gao WZ, Yang YH, Dan L, et al. Splenic tyrosine kinase promotes pulmonary angiogenesis in rats with hepatopulmonary syndrome[J]. 生理学报,2020, 72:785-792.
[8]Gao J, Yu H, Bai X, et al. Loss of cell polarity regulated by PTEN/Cdc42 enrolled in the process of Hepatopulmonary Syndrome[J]. J Cell Mol Med,2019,23:5542-5552.
[9]Yang Y, Yu H, Yang C, et al. Kruppel-like factor 6 mediates pulmonary angiogenesis in rat experimental hepatopulmonary syndrome and is aggravated by bone morphogenetic protein 9[J]. Biol Open,2019,8.doi: 10.1242/bio.040121.
[10]Ratcliffe CD, Sahgal P, Parachoniak CA,et al. Regula-tion of cell migration and beta1 integrin trafficking by the endosomal adaptor GGA3[J]. Traffic,2016, 17:670-688.
[11]Peng J, Li X, Zhang Y,et al. Par3/Integrin beta1 regulates embryo adhesion via changing endometrial luminal epithelium polarity[J]. Biol Reprod,2021, ioab033.doi: 10.1093/biolre/ioab033.
[12]Babel L, Kruse L, Bump S, et al. Lipid-rafts remain stable even after ionizing radiation induced disintegration of beta1 integrin containing focal adhesions[J]. BMC Res Notes,2017, 10:697. doi: 10.1186/s13104-017-3032-8.
[13]Antoniades I, Kyriakou M, Charalambous A, et al. FAK displacement from focal adhesions: a promising strategy to target processes implicated in cancer progression and metastasis[J]. Cell Commun Signal,2021, 19:3. doi: 10.1186/s12964-020-00671-1.
[14]郝礼森,张家琪,刘博,等. PTEN表达下调促进体外活化大鼠肝星状细胞黏附[J]. 基础医学与临床,2017,37:364-368.
[15]Yuh DY, Maekawa T, Li X,et al. The secreted protein DEL-1 activates a beta3 integrin-FAK-ERK1/2-RUNX2 pathway and promotes osteogenic differentiation and bone regeneration[J]. J Biol Chem,2020, 295:7261-7273.