生物医学工程学杂志

生物医学工程学杂志

血根碱对大鼠气道平滑肌细胞生物力学特性的影响

查看全文

本研究拟探究血根碱对大鼠气道平滑肌细胞(rASMCs)的刚度、牵张力、应力纤维分布等细胞生物力学特性的影响。体外培养的 rASMCs 经过血根碱溶液在不同浓度(0.005~5 μmol/L)条件下分别处理 12 h、24 h、36 h 和 48 h 后,采用噻唑盐比色法、光学磁粒扭转细胞测量仪、傅里叶变换牵张力显微术、划痕实验和免疫荧光显微技术等检测其活性、刚度、牵张力、迁移速度和微丝骨架分布等细胞生物力学特性的变化。实验结果显示,在浓度低于 0.5 μmol/L 时,血根碱对细胞活性没有明显影响,但对细胞生物力学特性呈现浓度和时间依赖性,具体表现为 rAMSCs 经 0.05 μmol/L 和 0.5 μmol/L 浓度的血根碱处理 12 h 和 24 h 后细胞刚度、细胞牵张力和细胞迁移速度均明显降低、细胞骨架应力纤维出现解聚。鉴于气道平滑肌细胞(ASMCs)生物力学特性在哮喘气道高反应性(AHR)中的关键作用,上述的实验结果提示,血根碱有可能通过改变气道平滑肌细胞生物力学特性而改善 AHR,进而为开发基于血根碱的气道松弛剂等哮喘治疗药物奠定基础。

This study aimed to evaluate the effect of sanguinarine on biomechanical properties of rat airway smooth muscle cells (rASMCs) including stiffness, traction force and cytoskeletal stress fiber organization. To do so, rASMCs cultured in vitro were treated with sanguinarine solution at different concentrations (0.005~5 μmol/L) for 12 h, 24 h, 36 h, and 48 h, respectively. Subsequently, the cells were tested for their viability, stiffness, traction force, migration and microfilament distribution by using methylthiazolyldiphenyl-tetrazolium bromide assay, optical magnetic twisting cytometry, Fourier transform traction microscopy, scratch wound healing method, and immunofluorescence microscopy, respectively. The results showed that at concentration below 0.5 μmol/L sanguinarine had no effect on cell viability, but caused dose and time dependent effect on cell biomechanics. Specifically, rASMCs treated with sanguinarine at 0.05 μmol/L and 0.5 μmol/L for 12 and 24 h exhibited significant reduction in stiffness, traction force and migration speed, together with disorganization of the cytoskeletal stress fibers. Considering the essential role of airway smooth muscle cells (ASMCs) biomechanics in the airway hyperresponsiveness (AHR) of asthma, these findings suggest that sanguinarine may ameliorate AHR via alteration of ASMCs biomechanical properties, thus providing a novel approach for asthma drug development.

关键词: 气道平滑肌细胞; 血根碱; 生物力学; 细胞刚度; 细胞牵张力

Key words: airway smooth muscle cells; sanguinarine; biomechanics; cell stiffness; cell traction force

引用本文: 罗明志, 余培丽, 金阳, 刘磊, 李晶晶, 潘艳, 邓林红. 血根碱对大鼠气道平滑肌细胞生物力学特性的影响. 生物医学工程学杂志, 2018, 35(4): 583-591. doi: 10.7507/1001-5515.201708025 复制

登录后 ,请手动点击刷新查看全文内容。 没有账号,
登录后 ,请手动点击刷新查看图表内容。 没有账号,
1. Mccracken J L, Veeranki S P, Ameredes B T, et al. Diagnosis and management of asthma in adults: a review. JAMA, 2017, 318(3): 279-290.
2. Rydell-Törmänen K, Risse P A, Kanabar V, et al. Smooth muscle in tissue remodeling and hyper-reactivity: airways and arteries. Pulm Pharmacol Ther, 2013, 26(1): 13-23.
3. Chapman D G, Irvin C G. Mechanisms of airway hyper-responsiveness in asthma: the past, present and yet to come. Clinical and Experimental Allergy, 2015, 45(4): 706-719.
4. Bates J H, Maksym G N. Mechanical determinants of airways hyperresponsiveness. Crit Rev Biomed Eng, 2011, 39(4): 281-296.
5. Ozier A, Allard B, Bara I, et al. The pivotal role of airway smooth muscle in asthma pathophysiology. J Allergy (Cairo), 2011: 742710.
6. Affonce D A, Lutchen K R. New perspectives on the mechanical basis for airway hyperreactivity and airway hypersensitivity in asthma. J Appl Physiol (1985), 2006, 101(6): 1710-1719.
7. Janssen L J. Airway smooth muscle as a target in asthma and the beneficial effects of bronchial thermoplasty. J Allergy (Cairo), 2012: 593784.
8. Camoretti-Mercado B. Targeting the airway smooth muscle for asthma treatment. Transl Res, 2009, 154(4): 165-174.
9. Zuyderduyn S, Sukkar M B, Fust A, et al. Treating asthma means treating airway smooth muscle cells. Eur Respir J, 2008, 32(2): 265-274.
10. Stephens N L, Li Weilong, Jiang He, et al. The biophysics of asthmatic airway smooth muscle. Respir Physiol Neurobiol, 2003, 137(2/3): 125-140.
11. Black J L, Panettieri R A, Banerjee A, et al. Airway smooth muscle in asthma: just a target for bronchodilation?. Clin Chest Med, 2012, 33(3): 543-558.
12. Oria R, Wiegand T, Escribano J, et al. Force loading explains spatial sensing of ligands by cells. Nature, 2017, 552(7684): 219-224.
13. Dinardo C L, Santos H C, Vaquero A R, et al. Smoking and female sex: independent predictors of human vascular smooth muscle cells stiffening. PLoS One, 2015, 10(12): e0145062.
14. Trepat X, DENG Linhong, An S S, et al. Universal physical responses to stretch in the living cell. Nature, 2007, 447(7144): 592-595.
15. Rosner S R, Pascoe C D, Blankman E, et al. The actin regulator zyxin reinforces airway smooth muscle and accumulates in airways of fatal asthmatics. PLoS One, 2017, 12(3): e0171728.
16. Deshpande D A, Wang W C H, Mcilmoyle E L, et al. Bitter taste receptors on airway smooth muscle bronchodilate by a localized Calcium flux and reverse obstruction. Nat Med, 2010, 16(11): 1299-1304.
17. Park C Y, Zhou E H, Tambe D, et al. High-throughput screening for modulators of cellular contractile force. Integr Biol (Camb), 2015, 7(10): 1318-1324.
18. Hu C M, Cheng H W, Cheng Y W, et al. Mechanisms underlying the induction of vasorelaxation in rat thoracic aorta by sanguinarine. Jpn J Pharmacol, 2001, 85(1): 47-53.
19. 王慧, 王悦尚, 刘兆颖, 等. 血根碱对大鼠肠平滑肌细胞收缩的抑制作用. 湖南农业大学学报:自然科学版, 2012, 38(05): 519-525.
20. Wang Hui, Yin G, Yu Chunhong, et al. Inhibitory effect of sanguinarine on PKC-CPI-17 pathway mediating by muscarinic receptors in dispersed intestinal smooth muscle cells. Res Vet Sci, 2013, 95(3): 1125-1133.
21. Wang Yue, Lu Yun, Luo Mingzhi, et al. Evaluation of pharmacological relaxation effect of the natural product naringin on in vitro cultured airway smooth muscle cells and in vivo ovalbumin-induced asthma Balb/c mice. Biomedical reports, 2016, 5(6): 715-722.
22. Zhang Ying, Ng S S, Wang Yilei, et al. Collective cell traction force analysis on aligned smooth muscle cell sheet between three-dimensional microwalls. Interface Focus, 2014, 4(2): 20130056.
23. An S S, Fabry B, Trepat X, et al. Do biophysical properties of the airway smooth muscle in culture predict airway hyperresponsiveness?. Am J Respir Cell Mol Biol, 2006, 35(1): 55-64.
24. Seow C Y. Passive stiffness of airway smooth muscle: the next target for improving airway distensibility and treatment for asthma?. Pulm Pharmacol Ther, 2013, 26(1): 37-41.
25. 贾海燕, 徐婉晴, 张庭华, 等. 血根碱对大鼠离体子宫平滑肌收缩的影响及其机制研究. 中国畜牧兽医, 2017, 44(07): 2209-2213.