载硫化氢气体超声微泡的制备及其性能评价

摘要/Abstract

摘要： 目的制备一种新型的载硫化氢气体的脂质微泡（HSMB）并评价其理化特性和超声显影能力。方法硫化氢和全氟丙烷
按4∶0、3∶1、2∶2、1∶3和0∶4混合后，机械震荡法制备5种载相应混合气体的HSMB，即HSMB4∶0、HSMB3∶1、HSMB2∶2、HSMB1∶3和
HSMB0∶4。检测微泡的粒径及浓度并评价其稳定性，优化气体混合比例。选取优化的混合气体制备的HSMB，观察超声辐照对
微泡浓度及下清液中硫化氢浓度的影响；观察大鼠心肌及肾脏的超声显影效果。结果HSMB外观呈乳白色，镜下呈球形结构，无
聚集现象。HSMB4∶0和HSMB3∶1微泡初始浓度较HSMB2∶2明显减少（P<0.05），且均随时间的延长逐渐降低。HSMB2∶2、HSMB1∶3
和HSMB0∶4微泡初始浓度无差别（P>0.05），72 h内浓度无变化（P>0.05）。超声辐照后，HSMB2∶2微泡浓度显著减少（P<0.05），而
下清液中的硫化氢浓度明显增加（P<0.05）。HSMB2∶2静脉注射后获得良好的心脏及肾脏超声显影效果。结论硫化氢和全氟丙
烷以2∶2混合制备的HSMB浓度高、稳定性良好，可在超声辐照下释放硫化氢，具有良好的超声显像效果，有望实现硫化氢气体
可视化靶向传输。

Abstract: Objective To prepare a phospholipid-coated microbubble loaded with hydrogen sulfide (HSMB) and evaluate its
physicochemical and acoustic properties. Methods Hydrogen sulfide and perfluoropropane were mixed at the ratios of 4:0, 3:
1, 2:2, 1:3, and 0:4 to prepare hydrogen sulfide-loaded microbubbles (termed HSMB4:0, HSMB3:1, HSMB2:2, HSMB1:3, and HSMB0:4,
respectively). The microbubble concentration and diameter were investigated and their stability were evaluated. The optimal
ratio of hydrogen sulfide and perfluoropropane was determined according to the changes of microbubble concentration. The
changes of dissolved hydrogen sulfide and concentration of the microbubbles were investigated after exposure to ultrasound,
and their acoustic enhancement effects in the myocardium and kidney were observed after intravenous injection in rats.
Results HSMBs were milky in color and spherical in shape without aggregations. The concentrations of HSMB4:0 and HSMB3:1
were lower than that of HSMB2:2 and decreased with time. HSMB2:2, HSMB1:3 and HSMB0:4 showed comparable concentrations
and were stable within 72 h. After exposure to ultrasound, the concentration of HSMB2:2 decreased while the dissolved
hydrogen sulfide increased significantly. Intravenous injection of HSMB2:2 produced a satisfactory contrast-enhancing effect in
the myocardium and kidney of rats. Conclusion HSMB prepared with the hydrogen sulfide to perfluoropropane ratio of 2:2
has excellent contrast-enhancing effect and is capable of carrying and releasing hydrogen sulfide upon ultrasound exposure to
potentially allow visual site-specific delivery of hydrogen sulfide.

陈钢彬，杨莉，李新忠，钟林涛，吴爵非，宾建平. 载硫化氢气体超声微泡的制备及其性能评价[J]. 南方医科大学学报, 2015, 35(12): 1672-.

CHEN Gangbin1, YANG Li2, LI Xinzhong1, ZHONG Lintao1, WU Juefei1, BIN Jianping1. Preparation and characterization of microbubbles loaded with hydrogen sulfide[J]. Journal of Southern Medical University, 2015, 35(12): 1672-.

参考文献

［1］ Polhemus DJ, Lefer DJ. Emergence of Hydrogen sulfide as an endogenous gaseous signaling molecule in cardiovascular disease ［J］. Circ Res, 2014, 114(4): 730-7.
［2］ Beauchamp Ro Jr, Bus JS, Popp J A, et al. A critical review of the literature on hydrogen sulfide toxicity［J］. Crit Rev Toxicol, 1984, 13 (1): 25-97.
［3］ Yoo D, Jupiter RC, Pankey EA, et al. Analysis of cardiovascular responses to the H2S donors Na2S and NaHS in the rat［J］. Am J Physiol Heart Circ Physiol, 2015, 309(4): H605-14.
［4］ Almeida AF, Guidotti TL. Differential sensitivity of lung and brain to sulfide exposure:a peripheral mechanism for apnea［J］. Toxicol Sci, 1999, 50(2): 287-93.
［5］ Fix SM, Borden MA, Dayton PA. Therapeutic gas delivery via microbubbles and liposomes［J］. J Control Release, 2015, 209: 139-49.
［6］ Tong JY, Ding JD, Shen XB, et al. Mesenchymal stem cell transplantation enhancement in myocardial infarction rat model under ultrasound combined with nitric oxide microbubbles［J］. PLoS One, 2013, 8(11): e80186.
［7］ Mcewan C, Owen J, Stride E, et al. Oxygen carrying microbubbles for enhanced sonodynamic therapy of hypoxic tumours［J］. J Control Release, 2015, 203: 51-6.
［8］谭开彬, 高云华, 刘平, 等. 机械振荡法制备脂膜超声造影剂的初步实验研究［J］. 中国超声医学杂志, 2006(8): 561-3.
［9］ Fujii H, Li SH, Wu J, et al. Repeated and targeted transfer of angiogenic plasmids into the infarcted rat heart via ultrasound targeted microbubble destruction enhances cardiac repair［J］. Eur Heart J, 2011, 32(16): 2075-84.
［10］Sato T, Mori S, Sakamoto M, et al. Direct delivery of a cytotoxic anticancer agent into the metastatic lymph node using nano/ microbubbles and ultrasound［J］. PLoS One, 2015, 10(4): e123619.
［11］Eisenbrey JR, Albala L, Kramer MR, et al. Development of an ultrasound sensitive Oxygen carrier for Oxygen delivery to hypoxic tissue［J］. Int J Pharm, 2015, 478(1): 361-7.
［12］Britton GL, Kim H, Kee PH, et al. In vivo therapeutic gas delivery for neuroprotection with echogenic liposomes［J］. Circulation, 2010, 122(16): 1578-87.
［13］Mcewan C, Owen J, Stride E, et al. Oxygen carrying microbubbles for enhanced sonodynamic therapy of hypoxic tumours［J］. J Control Release, 2015, 203: 51-6.
［14］Kwan JJ, Borden MA. Lipid monolayer collapse and microbubble stability［J］. Adv Colloid Interface Sci, 2012, 183: 82-99.
［15］Szijjarto C, Rossi S, Waton G, et al. Effects of perfluorocarbongases on the size and stability characteristics of Phospholipid- Coated microbubbles: osmotic effect versus interfacial film stabilization［J］. Langmuir, 2012, 28(2): 1182-9.
［16］Sutton JT, Raymond JL, Verleye MC, et al. Pulsed ultrasound enhances the delivery of nitric oxide from bubble liposomes to ex vivo porcine carotid tissue［J］. Int J Nanomedicine, 2014, 9: 4671-83.
［17］Kwan JJ, Kaya M, Borden MA, et al. Theranostic Oxygen delivery using ultrasound and microbubbles［J］. Theranostics, 2012, 2(12):1174-84.
［18］郑扬, 廖锋, 杜军保, 等. 改良亚甲基蓝法测定血浆硫化氢［J］. 生理学报, 2012(6): 681-6.
［19］Wang Y, Zhao X, Jin H, et al. Role of Hydrogen sulfide in the development of atherosclerotic lesions in apolipoprotein E knockout mice［J］. Arterioscler Thromb Vasc Biol, 2009, 29(2): 173-9.
［20］李芳, 曾欧, 罗健, 等. 硫化氢对糖尿病大鼠心肌纤维化及MAPK1/3和MMP-8表达的影响［J］. 南方医科大学学报, 2015(4): 549-52.
［21］Kondo K, Bhushan S, King AL, et al. H(2)S protects against pressure overload-induced heart failure via upregulation of endothelial nitric oxide synthase［J］. Circulation, 2013, 127(10): 1116-27.
［22］ Elrod JW, Calvert JW, Morrison J, et al. Hydrogen sulfide attenuates myocardial ischemia-reperfusion injury by preservation of mitochondrial function［J］. Proc Natl Acad Sci USA, 2007, 104 (39): 15560-5.