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浙江大学学报(医学版)  2022, Vol. 51 Issue (4): 430-437    DOI: 10.3724/zdxbyxb-2022-0178
专题报道     
激活红细胞2,3 -二磷酸甘油酸变位酶活性的中药组分筛选
郭茜文1,2,李文斌2,*,王荣2,李加忠3,张汝学2,*,张晓静2,王子晗1,2
1. 甘肃中医药大学药学院,甘肃 兰州 730000
2. 中国人民解放军联勤保障部队第九四〇医院药剂科 全军高原医学实验室,甘肃兰州 730050
3. 兰州大学药学院,甘肃 兰州 730000
Screening of activators of 2,3-diphosphoglycerate mutase from traditional Chinese herb medicines
GUO Qianwen1,2,LI Wenbin2,*,WANG Rong2,LI Jiazhong3,ZHANG Ruxue2,*,ZHANG Xiaojing2,WANG Zihan1,2
1. College of Pharmacy, Gansu University of Chinese Medicine, Lanzhou 730000, China;
2. Department of Pharmacy, the 940th Hospital of Joint Logistics Support Force of Chinese People’s Liberation Army, Key Laboratory of the Plateau Medicine, Lanzhou 730050, China;
3. School of Pharmacy, Lanzhou University, Lanzhou 730000, China
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摘要:

目的:从中药组分中筛选2,3-二磷酸甘油酸(BPG)变位酶(BPGM)的激活剂,以提高红细胞的缺氧耐受性。 方法:以红细胞胞浆蛋白BPGM为受体,常用中药成分数据库为配体,经过里宾斯基五规则类药性筛选后,采取刚性对接(LibDock)和半柔性对接(CDOCKER)进行虚拟筛选;提取红细胞胞浆蛋白,验证筛选出来的化合物对红细胞胞浆中BPGM亲和力的影响;最后,体外孵育红细胞,建立红细胞缺氧模型,并验证化合物对红细胞缺氧模型中BPGM活性的影响。 结果:通过刚性对接和半柔性对接优选出与BPGM结合能前十的化合物。用这十种化合物孵育胞浆蛋白,与空白对照组比较,迷迭香酸甲酯各剂量组、二氢姜黄素大剂量组、八氢姜黄素中剂量组和阿魏酸松柏酯大剂量组可以进一步激活BPGM,从而显著增加常氧红细胞中的2,3-BPG含量(均 P<0.05);四氢姜黄素小剂量组、橙黄胡椒酰胺大剂量和小剂量组、六氢姜黄素各剂量组和N-p-香豆酰-羟色胺中剂量组常氧红细胞2,3-BPG含量有增加的趋势(均 P>0.05)。缺氧红细胞在加入中剂量迷迭香酸甲酯、中剂量八氢姜黄素、大剂量六氢姜黄素和中剂量N-p-香豆酰-羟色胺后,2,3-BPG含量显著增加(均 P<0.05)。结论:迷迭香酸甲酯、八氢姜黄素、六氢姜黄素和 N-p-香豆酰-羟色胺可以激活BPGM从而提高缺氧红细胞中2,3-BPG含量。

关键词: 红细胞2,3-二磷酸甘油酸2,3-二磷酸甘油酸变位酶分子对接中药 组分    
Abstract:

Objective: To screen activators of 2,3-diphosphoglycerate (BPG) mutase (BPGM) from Chinese herb medicines, so as to improve the hypoxia tolerance of erythrocytes. Methods: BPGM was used as the receptor and Chinese medicine ingredients database was used as the ligand in the study. After Lipinski rule of five screening, LibDock and CDOCKER docking were used for virtual screening. The effect of the screened compounds on the affinity of BPGM in erythrocytes was verified. Finally, the erythrocytes were incubated in vitro to establish the erythrocyte hypoxia model, and the effect of the compound on the activity of BPGM in the erythrocyte hypoxia model was verified. Results: Ten compounds with highest binding affinity to BPGM were selected by LibDock and CDOCKER, and the cytoplasm protein was incubated with the ten compounds. Compared with blank control group, methyl rosmarinate group, dihydrocurcumin high dose group, octahydrocurcumin medium dose group and coniferyl ferulate high dose group were able to further activate the BPGM, significantly increase the levels of 2, 3-BPG in normal erythrocytes (all P<0.05); while the low dose of tetrahydrocurcumin, high dose and low dose of aurantiamide, hexahydrocurcumin and medium dose ofN- (p-coumaroyl) serotonin had a tendency to increase the contents of 2,3-BPG in normal erythrocytes (all P>0.05). In the hypoxic red blood cells, the medium dose methyl rosmarinate, medium dose octahydrocurcumin, high dose hexahydrocurcumin and medium doseN-(p-coumaroyl) serotonin could significantly increase the contents of 2,3-BPG (all P<0.05).Conclusion: The methyl rosmarinate, octahydrocurcumin, hexahydrocurcumin and N-(p-coumaroyl) serotonin could activate BPGM and increase the contents of 2,3-BPG in hypoxic erythrocytes.

Key words: Erythrocyte    2,3-Diphosphoglycerate    2,3-Diphosphoglycerate mutase    Molecular docking    Components of traditional Chinese medicine
收稿日期: 2022-04-18 出版日期: 2022-11-16
CLC:  R965.1  
基金资助: 国家自然科学基金(82173738)
通讯作者: 李文斌,张汝学   
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引用本文:

郭茜文,李文斌,王荣,李加忠,张汝学,张晓静,王子晗. 激活红细胞2,3 -二磷酸甘油酸变位酶活性的中药组分筛选[J]. 浙江大学学报(医学版), 2022, 51(4): 430-437.

GUO Qianwen,LI Wenbin,WANG Rong,LI Jiazhong,ZHANG Ruxue,ZHANG Xiaojing,WANG Zihan. Screening of activators of 2,3-diphosphoglycerate mutase from traditional Chinese herb medicines. J Zhejiang Univ (Med Sci), 2022, 51(4): 430-437.

链接本文:

https://www.zjujournals.com/med/CN/10.3724/zdxbyxb-2022-0178        https://www.zjujournals.com/med/CN/Y2022/V51/I4/430

图1  去水加氢后的BPGM蛋白(PDB ID: 3NFY)结构三维图 BPGM:2,3-二磷酸甘油酸变位酶.

序号

英文名称

中文名称

CAS编号

1

Trifolirhizin

三叶草素

6807-83-6

2

Yinyanghuo B

淫羊藿B

174286-24-9

3

Methylnissolin-3- O-glucoside

美迪紫檀苷

94367-42-7

4

Sophoraflavanone G

槐黄烷酮

97938-30-2

5

Deoxyrhapontin

去氧土大黄苷

30197-14-9

6

Sesamolin

芝麻林素

526-07-8

7

Octahydrocurcumin

八氢姜黄素

36062-07-4

8

Malabaricone C

马拉巴酮C

63335-25-1

9

Sesamin

芝麻素

607-80-7

10

paulownin

泡桐素

13040-46-5

11

Aurantiamide

橙黄胡椒酰胺

58115-31-4

12

Methyl rosmarinate

迷迭香酸甲酯

99353-00-1

13

Hexahydrocurcumin

六氢姜黄素

36062-05-2

14

Asarinin

细辛素

133-04-0

15

Malabaricone B

马拉巴酮B

63335-24-0

16

L-asarinin

L-细辛脂素

133-05-1

17

Malabaricone A

马拉巴酮A

63335-23-9

18

Neoglycyrol

新甘草酚

23013-84-5

19

Raspberry ketone glucoside

树莓苷

38963-94-9

20

Coniferyl ferulate

阿魏酸松柏酯

63644-62-2

21

Tetrahydrocurcumin

四氢姜黄素

36062-04-1

22

Curcumin

姜黄素

458-37-7

23

Drotaverine hydrochloride

盐酸屈他维林

985-12-6

24

N-(p-coumaroyl) serotonin

N-p-香豆酰-羟色胺

68573-24-0

25

Fargesin

辛夷脂素

31008-19-2

26

Coniferin

松柏醇葡萄糖苷

531-29-3

27

Epibrassinolide

表油菜素内酯

78821-43-9

28

Secoisolariciresinol

开环异落叶松树脂酚

29388-59-8

29

Dihydrocurcumin

二氢姜黄素

76474-56-1

30

Cabergoline

卡麦角林

81409-90-7

表1  与2,3-二磷酸甘油酸变位酶刚性对接LibDock 140分以上的化合物
图2  化合物与2,3-二磷酸甘油酸变位酶的分子对接模式图

组别

n

迷迭香酸甲酯

四氢姜黄素

橙黄胡椒酰胺

六氢姜黄素

二氢姜黄素

空白对照组

3

203.4±1.4

203.4±1.4

203.4±1.4

203.4±1.4

203.4±1.4

药物大剂量组

3

251.3±2.0 *

195.4±9.7

236.0±10.8

243.0±26.8

274.3±60.9 *

药物中剂量组

3

288.3±28.3 *

196.8±6.4

199.2±17.6

228.3±27.9

227.3±52.1

药物小剂量组

3

260.5±4.0 *

235.2±14.8

220.3±2.5

207.6±0.5

206.6±8.6

组别

n

N-p-香豆酰-羟色胺

姜黄素

八氢姜黄素

开环异落叶松树脂酚

阿魏酸松柏酯

空白对照组

3

203.4±1.4

203.4±1.4

203.4±1.4

203.4±1.4

203.4±1.4

药物大剂量组

3

199.5±11.0

193.8±25.1

210.5±24.1

193.8±25.1

262.7±59.6 *

药物中剂量组

3

217.1±12.0

189.4±10.1

274.5±12.9 *

189.4±10.1

207.65±57.8

药物小剂量组

3

190.0±18.5

177.8±7.8

214.2±33.1

177.8±7.8

211.9±32.2

表2  各组红细胞中2,3-二磷酸甘油酸含量

组别

谷胱甘肽含量(mg/gHb)

过氧化氢酶活(U/mgHb)

丙二醛含量(nmol/mgHb)

空白对照组

25.27±0.42

10.41±1.88

0.20±0.04

缺氧 15?min

17.17±2.88 *

7.59±1.28 *

0.28±0.02 *

缺氧 45?min

15.36±1.45 *

4.79±0.38 * #

0.29±0.01 *

缺氧 90?min

18.06±3.09 *

6.99±1.38 *△

0.25±0.02 *

缺氧 180?min

15.04±0.47 *

5.37±0.28 *#

0.27±0.03 *

表3  不同缺氧时间对红细胞谷胱甘肽含量、过氧化氢酶活性和丙二醛含量的影响
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