摘要：
BACKGROUND: Angelica sinensis and Aurantii fructu used in a pair, named Danggui-Zhiqiao herb-pair (DZHP), which was rich in essential oil and has been adopted to promote blood circulation, dispel blood stasis, and relieve pain in traditional Chinese medicine (TCM). OBJECTIVE: To analyze the composition and pharmacological effects of essential oil from DZHP. MATERIALS AND METHODS: The composition of the essential oil from DZHP was analyzed by gas chromatography/mass spectrometry (GC/MS). Its analgesic activity was evaluated by acetic acid-induced writhing test and hot plate test. The hemorheology test was carried out to evaluate the effect on hemorheology in rats with blood stasis syndrome. RESULTS: Twenty-eight components were identified and the main components were alpha-pinene (3.07%), beta-pinene (2.0%), beta-myrcene (3.71%), D-limonene (49.28%), gamma-terpinen (9.53%), alpha-terpinolene (1.80%), alpha-terpineol (2.02%), beta-bisabolene (1.13%), butylidenephthalide (1.43%), and Z-ligustilide (16.08%). The pharmacology test showed that the essential oil significantly inhibited the number of writhes induced by acetic acid with inhibition rate of 44.64% and significantly increased hot-plate latency compared with control group from 30 to 90 min after oral administration of drugs in mice. It could significantly decrease plasma viscosity, whole blood relative index at high and low shear rate, whole blood reduced viscosity at high and low shear rate, and erythrocyte rigidity index in hemorheology test. CONCLUSION: The composition of the essential oil of DZHP was determined successfully and it had analgesic and promoting blood circulation activities. SUMMARY: Angelica sinensis and Aurantii fructu used in a pair, named Danggui-Zhiqiao herb-pair (DZHP), which was rich in Essential oil and has been adopted to promote blood circulation, dispel blood stasis and relieve pain in traditional Chinese medicine (TCM).Twenty-eight components were identified and the main components were alpha-pinene (3.07%), beta-pinene (2.0%), beta-myrcene (3.71%), D-limonene (49.28%), gamma-terpinen (9.53%), alpha-terpinolene (1.80%), alpha-terpineol (2.02%), beta-bisabolene (1.13%), butylidenephthalide (1.43%), and Z-ligustilide (16.08%).The essential oil significantly inhibited the number of writhes induced by acetic acid and increased hot-plate latency compared with control group from 30 min to 90 min after oral administration of drugs in mice.The essential oil could significantly decrease plasma viscosity (PV), whole blood relative index (WBRI) at high and low shear rate, whole blood reduced viscosity (WBRV) at high and low shear rate, and erythrocyte rigidity index (ERI) in hemorheology test. Abbreviations used: DZHP: Danggui-Zhiqiao herb-pair; TCM: traditional Chinese medicine; GC/MS: gas chromatography /mass spectrometry; PV: plasma viscosity; WBRI: whole blood relative index; WBRV: whole blood reduced viscosity; ERI: erythrocyte rigidity index.
作者机构：
[宋厚盼; 李鑫; 蔡雄; 魏艳霞] Hunan Provincial Key Laboratory of Diagnostic and Therapeutic Research in Chinese Medicine, Hunan University of Chinese Medicine, Changsha, 410208, China;[王炜; Zeng R.; 李涛] School of Pharmaceutical Sciences, Hunan University of Chinese Medicine, Changsha, 410208, China;State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, 999078, Macau;[刘良] State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology999078, Macau;[余黄合; 李如意] Hunan Provincial Key Laboratory of Diagnostic and Therapeutic Research in Chinese Medicine, Hunan University of Chinese Medicine, Changsha, 410208, China, School of Pharmaceutical Sciences, Hunan University of Chinese Medicine, Changsha, 410208, China
作者机构：
[Song, Hou-Pan; Cai, Xiong; Huang, Hui-Yong] Hunan Univ Chinese Med, Inst TCM Diagnost, Changsha 410208, Hunan, Peoples R China.;[Song, Hou-Pan; Li, Ru-Liu] Guangzhou Univ Chinese Med, Spleen & Stomach Inst, Guangzhou 510405, Guangdong, Peoples R China.;[Zhou, Chi] Guangzhou Univ Chinese Med, Affiliated Hosp 1, Guangzhou 510405, Guangdong, Peoples R China.;[Song, Hou-Pan] Hunan Univ Chinese Med, Inst TCM Diagnost, 300 Xueshi Rd, Changsha 410208, Hunan, Peoples R China.
通讯机构：
[Song, Hou-Pan] H;Hunan Univ Chinese Med, Inst TCM Diagnost, 300 Xueshi Rd, Changsha 410208, Hunan, Peoples R China.
关键词：
Atractylodes macrocephala Koidz;Intestinal epithelial cells;Cell migration;Polyamines;Rho GTPases;Non-muscle myosin II
摘要：
Ethnopharmacological relevance: Atractylodes macrocephala Koidz (AMK), a valuable traditional Chinese herbal medicine, has been widely used in clinical practice for treating patients with disorders of the digestive system. AMK has shown noteworthy promoting effect on improving gastrointestinal function and immunity, which might represent a promising candidate for the treatment of intestinal mucosa injury. The aim of this study was to investigate the efficacy of AMK on intestinal mucosal restitution and the underlying mechanisms via intestinal epithelial (IEC-6) cell migration model. Materials and methods: A cell migration model of IEC-6 cells was induced by a single-edge razor blade along the diameter of the cell layers in six-well polystyrene plates. After wounding, the cells were grown in control cultures and in cultures containing spermidine (5 mu M, SPD, reference drug), alphadifluoromethylornithine (2.5 mM, DFMO, polyamine inhibitor), AMK (50, 100, and 200 mg/L), DFMO plus SPD and DEMO plus AMK for 12 h. The polyamines content was detected by high-performance liquid chromatography (HPLC) with pre-column derivatization. The Rho mRNAs expression levels were assessed by Q-RT-PCR. The Rho and non-muscle myosin II proteins expression levels were analyzed by Western blot. The formation and distribution of non-muscle myosin II stress fibers were monitored with immunostaining techniques using specific antibodies and observed by confocal microscopy. Cell migration assay was carried out using inverted microscope and the Image-Pro Plus software. All of these indexes were used to evaluate the effectiveness of AMK. Results: (1) Treatment with AMK caused significant increases in cellular polyamines content and Rho mRNAs and proteins expression levels, as compared to control group. Furthermore, AMK exposure increased non-muscle myosin II protein expression levels and formation of non-muscle myosin II stress fibers, and resulted in an acceleration of cell migration in IEC-6 cells. (2) Depletion of cellular polyamines by DEMO resulted in a decrease of cellular polyamines levels. Rho mRNAs and proteins expression, nonmuscle myosin II protein formation and distribution, thereby inhibiting IEC-6 cell migration. AMK not only reversed the inhibitory effects of DFMO on the polyamines content, Rho mRNAs and proteins expression, non-muscle myosin II protein formation and distribution, but also restored cell migration to control levels. Conclusions: The results obtained from this study revealed that AMK significantly stimulates the migration of IEC-6 cells through a polyamine dependent mechanism, which could accelerate the healing of intestinal injury. These findings suggest the potential value of AMK in curing intestinal diseases characterized by injury and ineffective repair of the intestinal mucosa in clinical practice. (C) 2014 Elsevier Ireland Ltd. All rights reserved.
