不同分子量海蚌类肝素理化性质分析及抗血栓活性评价

doi:10.11978/2022015

热带海洋学报 ›› 2023, Vol. 42 ›› Issue (1): 152-160.doi: 10.11978/2022015CSTR: 32234.14.2022015

不同分子量海蚌类肝素理化性质分析及抗血栓活性评价

陈观兰¹(), 陈建平¹, 李瑞¹, 贾学静¹, 刘晓菲¹, 宋兵兵¹, 钟赛意¹^,²^,³()

1.广东省水产品加工与安全重点实验室, 广东省海洋生物制品工程实验室, 广东省海洋食品工程技术研究中心(广东海洋大学食品科技学院), 广东湛江 524008
2.广东海洋大学深圳研究院, 广东深圳 518108
3.海洋食品精深加工关键技术省部共建协同创新中心(大连工业大学), 辽宁大连 116034

收稿日期:2022-01-26 修回日期:2022-03-29 出版日期:2023-01-10 发布日期:2022-03-25
通讯作者: 钟赛意。email: zsylxc@126.com
作者简介:
陈观兰(1994—), 女, 广东省惠州市人, 硕士研究生, 从事食品科学与工程。email: cgl202112@163.com
基金资助:
广东省重点领域研发计划项目(2020B1111030004); 国家重点研发计划重点专项(2019YFD0902005); 深圳市科技计划项目(JCYJ20170818111335796); 深圳市科技计划项目(PT202001-17); 湛江市科技计划项目(2019A01015); 广东省高校科技创新团队项目(2021KCXTD021)

Physicochemical analysis and evaluation of the antithrombotic activity of different molecular weights of heparin from clam Coelomactra antiquata

CHEN Guanlan¹(), CHEN Jianping¹, LI Rui¹, JIA Xuejing¹, LIU Xiaofei¹, SONG Bingbing¹, ZHONG Saiyi¹^,²^,³()

1. Guangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Province Engineering Laboratory for Marine Biological Products, Guangdong Provincial Engineering Technology Research Center of Seafood (College of Food Science and Technology, Guangdong Ocean University), Zhanjiang 524008, China
2. Shenzhen Research Institute, Guangdong Ocean University, Shenzhen 518108, China
3. Collaborative Innovation Center of Seafood Deep Processing, Dalian Polytechnic University, Dalian 116034, China

Received:2022-01-26 Revised:2022-03-29 Online:2023-01-10 Published:2022-03-25
Contact: ZHONG Saiyi. email: zsylxc@126.com
Supported by:
Guangdong Key Area Research and Development Program Project(2020B1111030004); National Key Research and Development Program Key Special Project(2019YFD0902005); Shenzhen Science and Technology Program Project(PT202001-17); Zhanjiang Science and Technology Program Project(2019A01015); Guangdong Province High School Science and Technology Innovation Team Project(2021KCXTD021); Shenzhen Science and Technology Program Project(JCYJ2017081818111335796)

摘要/Abstract

摘要：

为探究不同分子量海蚌类肝素的理化性质、体内抗血栓活性及对出血时间的影响, 本文以海蚌类肝素G2(60.25kDa) 及其 2 种不同分子量的降解产物DG1(24.48kDa)、DG2(6.75kDa) 为研究对象, 首先采用改良的硫酸-咔唑法和BaCl₂-gel 比浊法测定糖醛酸及硫酸基的含量, 利用纳米粒径仪测定粒径、Zeta电位, 并同步利用热分析仪分析热稳定性; 然后通过小鼠黑尾模型和小鼠断尾模型分别研究抗血栓活性及对出血时间的影响。研究结果表明, 随着分子量的降低, 海蚌类肝素硫酸基含量升高, 糖醛酸含量降低, 初始降解温度先升高后降低, 粒径降低, Zeta 电位升高。相对于海蚌类肝素G2, 其适度降解产物DG1具有更高的抑制尾部血栓形成的能力, 同时具有较低的出血风险。

关键词: 海蚌类肝素, 理化性质, 抗血栓, 出血副作用

Abstract:

To investigate the physicochemical properties in vivo antithrombotic activity and their effects on the bleeding time of different molecular weights of clam heparinoid, native clam heparinoid G2 (60.25 kDa), and its two different degradation products, DG1 (24.48 kDa) and DG2 (6.75 kDa), were taken as the research objects in this study. The content of glucuronic acid and the sulfate group was respectively determined by the modified sulfuric acid-carbazole method and BaCl₂-gel turbidimetric method, the particle size and zeta potential were determined by nanoparticle sizer, and the thermal stability was analyzed by the simultaneous thermal analyzer. Then the antithrombotic activity and the effect on bleeding time were investigated by the mice black tail model and mice broken tail model, respectively. The results showed that as the molecular weight of clam heparinoid decreased, the sulfate content increased, the glucuronic acid content decreased, the initial degradation temperature increased and then decreased, the particle size decreased, and the Zeta potential increased. The moderate degradation product of G2, DG1, has a higher ability to inhibit tail thrombosis with a lower risk of hemorrhage compared to the natural clam heparinoid G2.

Key words: clam heparinoid, physicochemical analysis, antithrombotic activity, bleeding time

陈观兰, 陈建平, 李瑞, 贾学静, 刘晓菲, 宋兵兵, 钟赛意. 不同分子量海蚌类肝素理化性质分析及抗血栓活性评价[J]. 热带海洋学报, 2023, 42(1): 152-160.

CHEN Guanlan, CHEN Jianping, LI Rui, JIA Xuejing, LIU Xiaofei, SONG Bingbing, ZHONG Saiyi. Physicochemical analysis and evaluation of the antithrombotic activity of different molecular weights of heparin from clam Coelomactra antiquata[J]. Journal of Tropical Oceanography, 2023, 42(1): 152-160.

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样品名称	硫酸基含量/%	糖醛酸含量/%
G2	18.07±1.28	27.17±1.57
DG1	24.74±0.64	22.38±0.93
DG2	27.33±2.22	15.32±3.23

样品名称	粒径/nm	PDI	Zeta电位/mV
G2	262.3±5.3	0.467±0.0218	-31.8±1.6
DG1	183.9±6.2	0.429±0.0339	-25.1±1.0
DG2	152.8±10.6	0.444±0.0169	-17.9±0.6

组别	W₀/g	W₇/g
B	25.3±3.4	32.8±2.5*
M	23.8±3.0	32.9±2.6*
Y	24.6±2.5	34.6±3.5*
G2L	24.6±3.5	33.4±1.9*
G2M	25.6±3.1	34.0±2.6*
G2H	24.5±2.9	33.6±1.9*
DG1L	24.0±3.0	34.8±2.2*
DG1M	23.6±2.2	35.6±2.7*
DG1H	24.4±3.3	33.0±2.4*
DG2L	24.4±3.5	33.6±3.0*
DG2M	23.9±3.8	32.1±1.8*
DG2H	23.9±3.2	31.6±3.4*

组别	黑尾比率/%
组别	24h	48h
B	0±0	0±0
M	52.03±5.46##	57.73±8.89##
Y	28.92±0.56**	33.01±1.61**
G2L	48.67±2.78	45.39±4.43
G2M	35.01±1.41**	33.32±2.09**
G2H	31.30±3.23**	30.83±4.34**
DG1L	46.91±5.27	45.05±3.88
DG1M	36.96±3.19*	34.31±1.68**
DG1H	19.94±5.26**	18.70±2.82**
DG2L	47.82±5.67	49.16±4.71
DG2M	47.30±4.70	47.25±6.61
DG2H	43.95±6.54	45.00±5.42

不同分子量海蚌类肝素理化性质分析及抗血栓活性评价

Physicochemical analysis and evaluation of the antithrombotic activity of different molecular weights of heparin from clam Coelomactra antiquata

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