In Vivo Antithrombotic Activity of Constituents from the Leaves of Craraegus pinnatifida Bge

Crataegus pinnatifida Bge (Hawthorn), well-known as both a medicine and food material, is the common name of the plants in the genus Crataegus of the Rosaceae family, which are distributed in East Asia, Europe and North America [1]. The greatest clinical use of hawthorn was in the treatment of various heart and cardiovascular ailments in Europe during the late 19th through to the early 20th centuries. Currently, hawthorn leaves, flowers, and both green (unripe) and red (ripe) berries are used to make herbal preparations to treat patients with severe heart disease: such as congestive heart failure, high blood pressure, hypoxia and hyperlipemia [2]. In order to evaluate the health protection value of hawthorn, the leaves of hawthorn were subjected to phytochemical investigation. By means of various chromatographic methods such as silica gel, sephadex LH-20, ODS column chromatography and semi-preparative HPLC, 98 compounds were isolated from the 80% EtOH extracts of the leaves of hawthorn, and their structures were elucidated on the basis of spectroscopic data and physico-chemical methods, such as 1D, 2D NMR, CD, HRESI-MS and chemical analysis. These compounds include 33 monoterpenes, 6 sesquiterpenes, 2 diterpenes, 11 triterpenes, 1 amide, 23 phenols, 8 lignans, 9 flavones, 3 hexenes, 2 sterols. Among them, 17 new compounds were named as pinnatifidanoside A-Q (1-17), 41 known compounds (18–59) were obtained from genus Crataegus for the first time. The antithrombotic activity of the isolated 34 compounds was investigated in a transgenic zebra fish system.

Table 1: Antithrombotic activity of 34 compounds on transgenic zebrafish system.

Number	Thrombosis (min)	Number	Thrombosis (min)	Number	Thrombosis (min)
Control 1	11.39±0.12	Control 2	15.12±0.07	Control 3	12.74±0.07
9	13.19±0.23	62	15.39±0.14	8	10.13±0.07
22	12.77±0.05	16	8.43±0.16	1	8.17±0.15
15	11.35±0.07	12	8.20±0.17	18	10.75±0.12
5	10.71±0.12	13	9.62±0.06	14	11.37±0.15
42	9.71±0.07	17	9.60±0.44	11	10.60±0.13
38	10.57±0.12	3	7.95±0.29	20	10.06±0.13
57	9.82±0.15	41	10.12±0.10	4	8.39±0.26
55	19.04±0.10	10	9.80±0.16	23	9.16±0.28
19	15.55±0.13	2	7.32±0.24	25	8.22±0.21
		21	7.65±0.13	39	7.63±0.23
		40	11.68±0.13	26	9.48±0.27
		24	13.47±0.20
		46	14.54±0.12

Acknowledgements: Thanks go to the Professor Qi-Shi Sun (Shenyang Pharmaceutical University, Shenyang, People's Republic of China) for identification of the plant material. References: [1] Zhang ZS, Ho WKK, et al. (2002) Food Res. International 35(9): 885–891. [2] Ding XP, Wang XT, et al. (2010) Food Chem 120: 929.