Volume 6,Issue 2,2014 Table of Contents

1 A Key Model: Translational Medicine in Research of Herbal Medicines

Chang-xiao Liu

2014, 6(2).

[Abstract](765) [HTML](0) [PDF 142.34 K](1736)

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It is delighted to read the paper of review titled “Correlation Analysis between Visceral Manifestation Theories on Xuanfa and Effect of Adrenergic Receptors” (CHM 2014, 5: 85-92) written by Professor Gang Bai et al at College of Pharmacy, Nankai University. The purpose of this article is very clear, as the authors said “The research and development of modern drugs should fully expect the guidance from Chinese visceral manifestation theory, and the application of this principle will guide the prevention and clinical treatment of a variety of refractory diseases caused by a change in environment, climate, or lifestyle”.

2 Correlation Analysis between Visceral Manifestation Theories on Xuanfa and Effect of Adrenergic Receptors

Gang Bai , Yuan-yuan Hou , Min Jiang , Jie Gao

2014, 6(2):85-92. DOI: 10.1016/S1674-6384(14)60013-7

[Abstract](1039) [HTML](0) [PDF 815.37 K](2382)

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Chinese visceral manifestation theory states that lung dominates qi, regulates breathing, and governs Xuanfa (dispersing) and Sujiang (descending). Clarifying this theory with modern physiological and pathological knowledge has been considered as an important part of complementary and alternative medicine therapy. Previous studies found that most Xuanfa drugs contained pharmacodynamic ingredients related to adrenergic receptors (ARs) signal transduction. The association of Xuanfa, with the control of breath movement, nutrient transfer, spreading heat to regulate temperature, and helping the heart control blood circulation, coincides with the physiological function of organs dominated by ARs-regulated sympathetic postganglionic fibers. Therefore, we hypothesize that Xuanfa is closely related to ARs-regulated signal transduction. By modern biological knowledge, we tried to evaluate and expound the correlation between the molecular mechanisms of modern physiology or pathology and Xuanfa or Sujiang theory. Ultimately, the research and development of modern drugs should fully expect the guidance from Chinese visceral manifestation theory, and the application of this principle will guide the prevention and clinical treatment of a variety of refractory diseases caused by a change in environment, climate, or lifestyle.

3 Traditional Chinese Medicine for Diminished Ovarian Reserve: A Systematic Review and Meta-analysis

Tian Xia , Rui-hong Ma , Wei Mu , Meng-di Hu , Sai-hua Ma , Shu-wen Mao , Yu Fu

2014, 6(2):93-102. DOI: 10.1016/S1674-6384(14)60014-9

[Abstract](1461) [HTML](0) [PDF 358.86 K](2358)

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Objective To assess the effectiveness and safety of traditional Chinese medicine (TCM) for women with diminished ovarian reserve (DOR). Methods A literature search was conducted in eight electronic databases for randomized controlled trials. Results Seventeen randomized controlled trials involving 1174 patients were included. Meta-analysis indicated that TCM was superior to Western medicine (WM) in reducing basal serum FSH level [MD = -1.70, 95% CI (-2.63, -0.77); P = 0.0004] and FSH/LH (MD = -0.43, 95% CI [-0.56, -0.30]; P = 0.0001), and the effect was more obvious two months after the last treatment (MD = -4.60, 95% CI [-6.26, -2.90], P < 0.000 01 and MD = -0.56, 95% CI [-0.85, -0.28], P = 0.0001), and increasing antral follicle count (AFC) (MD = 0.44, 95% CI [0.04-0.83]; P = 0.03). The review also revealed the positive role of CMM as an adjuvant to IVF-ET in improving pregnancy rate (PR = 1.75, 95% CI [1.25, 2.46]; P = 0.001). Conclusion TCM, with its unique way of replenishing the kidney, may provide an effective and safe alternative therapy to patients with DOR.

4 Antihypertensive Effects of Extract from Flower Buds of Coreopsis tinctoria on Spontaneously Hypertensive Rats

Qiang Yang , Yu-hua Sun , Li Zhang , Lei Xu , Meng-ying Hu , Xiao-yan Liu , Feng-ying Shi , Zheng-yi Gu

2014, 6(2):103-109. DOI: 10.1016/S1674-6384(14)60015-0

[Abstract](752) [HTML](0) [PDF 580.33 K](2808)

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Objective To provide experimental evidence for the antihypertensive activity of the ?avonoids in flower buds of Coreopsis tinctoria (CT-F) and to investigate the underlying mechanism. Methods The spontaneously hypertensive rats (SHRs) were divided into model, captopril (positive control), and CT-F groups, and the Wistar-Kyoto rats were set as control group, eight in each group. The blood pressure of SHRs, the activity of angiotensin II (Ang-II) in plasma, nitric oxide (NO), malondialdehyde (MDA) and thoracic aorta media thickness in SHRs were measured by tail-cuff method, radioimmunity method, nitrate reductase method, thibabituric acid (TBA) method, and the hematoxylin-eosin staining method. Q-PCR analysis was performed to determine the relative quantity of angiotensin-converting enzyme (ACE), ACEII, angiotensin type 1 receptor (AT1R), and TGF-β1 mRNA in left ventricle. Results CT-F could lower the systolic blood pressure of SHRs dramatically (P < 0.01). The levels of MDA in serum and Ang II in plasma of SHRs treated with CT-F decreased markedly (P < 0.05, 0.01), the level of NO in serum increased significantly (P < 0.01). In addition, thoracic aorta media thickness in SHRs treated with CT-F was thinner than that of the model group (P < 0.05). The mRNA expression of ACE, AT1R, and TGF-β1 in left ventricle was markedly decreased (P < 0.05), while that of ACE II was increased (P < 0.05). Conclusion CT-F is effective to lower the blood pressure of SHRs, and its antihypertensive effect is probably associated with lowering the oxidative stress by reducing MDA, ameliorating aorta remodeling, dilating vessel by increasing NO and decreasing Ang-II, and regulating the expression of rennin-angiotensin System-related various genes.

5 Effects of Puerarin on Experimental Model of Retinal Vein Occlusion in Rats

Chun-hua Hao , Wei-ting Wang , Ming-xiang Shao , Zhuan-you Zhao , Li-da Tang

2014, 6(2):110-114. DOI: 10.1016/S1674-6384(14)60016-2

[Abstract](690) [HTML](0) [PDF 370.73 K](2569)

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Objective To establish a model of retinal vein occlusion (RVO) in rats and to study the effect of puerarin on ischemic retinal disease and the corresponding mechanism. Methods RVO was induced in 10 adult male Sprague-Dawley (SD) rats by laser photothrombosis. Retinal blood flow was examined before and after 1 h of the operation, and model rats with the retinal vein blood flow decreasing by 50% compared to the basic value were chosen and then puerarins (20, 40, and 80 mg/kg) were given. The levels of vascular endothelial growth factor (VEGF), interleukin-1β (IL-1β), and nitric oxide (NO) were analyzed. In addition, the histopathology of RVO-eyes was performed. Results RVO-eyes displayed the signs of retinal damage and ischemia on Doppler Flowmeter and histopathology. Puerarin (20, 40, and 80 mg/kg) increased blood flow by 9.3% (P < 0.05), 33.1% (P < 0.001), and 41.5% (P < 0.001)， respectively. On the other hand, the histological changes were less severe at different degrees, relieving the symptoms such as edematous and thick neuroretinal layers, lax, edematous, and disorganized optic fibers layers, swollen and confused inner and outer nuclear layer. Besides, dose-dependent decrease of VEGF and IL-1β and increase of NO in vitreous fluid were observed, with respect to the model group. Conclusion A rat model of laser photochemical-induced RVO is established and a decrease in the retinal blood flow and histological damage is detected. The puerarin has therapeutic benefit in the rat model of RVO, through the pathway of neovascularization, anti-inflammation, and increase of NO.

6 Anti-oxidative Activities of Ethanol Extracts from Both Wild Plant and Suspension Cell Cultures of Rheum franzenbachii

Jun-li Wang , Ming-fei Liu , Xiao-xu Li , Qian Wang , Lu Zhang , Jian-fei Li

2014, 6(2):115-119. DOI: 10.1016/S1674-6384(14)60017-4

[Abstract](746) [HTML](0) [PDF 314.62 K](2213)

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Objective To evaluate the content of rhaponticin and anti-oxidative activities of the ethanol extracts from both the wild plants and suspension cell cultures of Rheum franzenbachii. Methods Quantitative analysis of rhaponticin was performed by HPLC. The anti-oxidative activities of the ethanol extracts were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging assays. Results The content of rhaponticin in the roots of the wild plant was 4.36 mg/g, while the content was only 1.59 mg/g in the leaves. The content of rhaponticin in suspension cells cultured on Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 6-benzylaminopurine (6-BAP) and 2.0 mg/L 2,4-dicholorophenoxy acetic acid (2,4-D) was 17.64 mg/g, which increased by 4.05 times compared with the content in the roots of the wild plants. The roots of wild plants displayed the strongest anti-oxidative activity, followed by the suspension cells 5 and 6, and the scavenging percent was 91.96%, 91.23%, and 89.27%, respectively, at the concentration of 100 μg/mL. The IC50 values were 2.477, 15.644, and 31.415 μg/mL, respectively. In particular, the DPPH scavenging activity of the ethanol extracts from the roots of the wild plant was generally comparable to the control of ascorbic acid (VC), and the IC50 value of the extracts was lower than that of VC (2.502 μg/mL). Conclusion Rhaponticin production in the cell culture can be modulated and the accumulation can be increased. The roots of the wild plant display the strongest anti-oxidative activity. These results suggest that R. franzenbachii could hold a good potential source for human health.

7 Effects of Sijunzi Dripping Pill on Gastrointestinal Motility of Mice

Ying-li Wang , Yan Wang

2014, 6(2):120-124. DOI: 10.1016/S1674-6384(14)60018-6

[Abstract](1029) [HTML](0) [PDF 301.57 K](1757)

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Objective To study the effects of Sijunzi Dripping Pill (SDP) on gastrointestinal motility of mice. Methods The diarrhea and swimming model of mice was made by Rhei Radix et Rhizoma-induced spleen deficiency. The intestinal transit, gastric emptying test, serum motilin (MTL), vasoactive intestinal peptide (VIP), and substance P (SP) were chosen to observe the effects of high-, mid-, and low-dose SDP on stomach movements, and the water extractive of Sijunzi Decoction was used as positive control. Results Compared with the control group, the gastric emptying rate in the gastrointestinal motility group was significantly decreased, the intestinal propulsion rate was obviously increased, the levels of MTL, prostaglandin E2 (PGE2), and SP were increased (P < 0.05), while the level of VIP was decreased (P < 0.05). Compared with the model group, SDP could decrease the intestinal transit rate, whereas increase the gastric emptying rate and the level of MTL (P < 0.05); The high-dose SDP could decrease the level of PGE2 (P < 0.05) and the low-dose SDP could decrease the level of VIP (P < 0.05); Each group had no significant effect on SP. Conclusion SDP has the good effect on increasing the gastrointestinal motility of mice, and its function may partly relate to the regulation of the levels of MTL and VIP as well as PGE2.

8 Determination of Protopine in Rat Brain Tissues by RRLC-ESI/Q-TOF-MS Method

Pei-xiang Wang , Yong-hui Li , Yan-jing Li , Ting Geng , Ming-li Li , Gang Ding , Zhen-zhong Wang , Wei Xiao

2014, 6(2):125-130. DOI: 10.1016/S1674-6384(14)60019-8

[Abstract](719) [HTML](0) [PDF 319.98 K](2001)

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Objective To analyse the quantification of protopine from Corydalis Decumbentis Rhizoma (CDR) extract in brain tissues of rats. Methods A rapid, sensitive, and accurate analytical method based on rapid resolution liquid chromatography (RRLC) coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (Q-TOF-MS) was developed for the quantification of protopine in brain of rats. A simple liquid-liquid extraction process was employed for the sample preparation. Chromatographic separation was achieved using 1.8 μm porous particle columns. Results The calibration curve of protopine was linear in the range of 12–897 ng/mL. The relative standard deviations of intra- and inter-day precision values were less than 10%. The extraction recoveries were 96.4%, 99.6%, and 98.5%, for protopine at the concentration of 598.0, 119.6, and 12.0 ng/mL, respectively, and internal standard (1.27 μg/mL) was (98.60 ± 0.02)%. Conclusion The validated method is successfully applied for the determination of protopine in brain tissue of rats after ig administration of CDR extract.

9 Research on Hypnotic and Anticonvulsant Activities of Total Alkaloids in Leaves of Eucommia ulmoides

Xin Li , Li-peng Yang , Xin-ling Li , Li-ping Wang , Shao-yang Liu , Yan Liu

2014, 6(2):131-135. DOI: 10.1016/S1674-6384(14)60020-4

[Abstract](1052) [HTML](0) [PDF 261.34 K](1873)

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Objective To study the hypnotic and anticonvulsant activities of the leaves of Eucommia ulmoides (LEU), the total alkaloids were extracted from LEU using water-acid method. Methods Mice were divided into five groups, including groups contained total alkaloids in three different doses, negative and positive control groups. Direct hypnotic experiment in mice, pentobarbital sodium synergistic experiment, and anticonvulsant experiment were used. The numbers of spontaneous activities, sleep rate, sleep latency, sleep time, convulsion rate, and convulsion latency of the mice were recorded and analyzed. Results The total alkaloids from LEU could increase the sleep rate, significantly lengthen the sleep time, and shorten the sleep latency of mice. Even the low dose of total alkaloids (0.33 g/kg) showed significantly different activities with negative control group (physiological saline) with the synergistic effect of the superthreshold dose of pentobarbital sodium. Furthermore, the total alkaloids efficiently inhibited the convulsion caused by nikethamide. Conclusion The total alkaloids from LEU have the excellent sedative, hypnotic, and anticonvulsant activities in mice, with high safety and little drug side effects. Therefore, they have the protential development prospects in sedative-hypnotic drugs.

10 Antihyperglycemic Effect of Various Fractions from Residues of Blumea balsamifera

Yan Xia , Jian Zuo , Xiang Li , Jian-wei Chen

2014, 6(2):136-139. DOI: 10.1016/S1674-6384(14)60021-6

[Abstract](971) [HTML](0) [PDF 299.35 K](1844)

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Objective To evaluate the antihyperglycemic activities of various fractions from the residues of Blumea balsamifera (BB), and to properly utilize the waste resource. Methods The antihyperglycemic activities were evaluated by the suppression on serum glucose level in vivo and α-glucosidase inhibition assay in vitro. The high-, mid-, and low-dose (1, 0.5, and 0.25 g/kg of the herb) fractions were ig given to mice for 8 d. The serum glucose was monitored at 1 and 12 h after feeding. Results The fasting and postprandial serum glucose levels of mice treated with high-dose petroleum ether fraction, ethyl acetate fraction, butyl alcohol fraction, methanol fraction, and water extract from BB were 4.45, 4.39, 4.43, 4.15, 3.74 mmol/L and 6.98, 6.23, 6.45, 6.26, 5.88 mmol/L, respectively, while those in vehicle control group were 5.63 and 7.50 mmol/L. There are four different inhibiting manners by the results of α-glucosidase inhibition assay. Conclusion The residues of BB have anti-diabetes activities after steam distillation.

11 Purification of Puerarin from Pueraria lobata by FCPC versus HSCCC Using Small-volume Columns

Bai-ling Sun , Yi-fang Yang , Xiao Hu , Xin-xin Xie , Jérémy Meucci , Ming Fang , Li-juan Zhao

2014, 6(2):140-144. DOI: 10.1016/S1674-6384(14)60022-8

[Abstract](817) [HTML](0) [PDF 292.63 K](1983)

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Objective To develop an efficient method for separating and purifying puerarin from the roots of Pueraria lobata. Methods Separation by fast centrifugal partition chromatography (FCPC) was processed with a biphasic solvent system composed of ethyl acetate-n-butanol-water (2:1:3). The separation conditions were determined as follows: sample loading of 10 mg, flow rate of 2 mL/min, rotation speed of 2200 r/min, ascending mode, and detection wavelength of 254 nm. High speed countercurrent chromatography (HSCCC) was used as a comparative method with the rotation speed of 800 r/min, flow rate of 2.0 mL/min, and sample loading of 10 mg in tail-to-head mode. Results Puerarin was obtained by FCPC with a resolution of 0.90 and a purity above 99%, while a resolution below 0.50 and a purity below 90% by HSCCC. Compared with HSCCC, FCPC has the advantages with higher purity and better resolution. Conclusion FCPC is a powerful method to separate and purify puerarin.

12 Changes of Secondary Metabolites and Trace Elements in Gentiana macrophylla Flowers: A Potential Medicinal Plant Part

Xiao-xue Niu , Xiao-wen Chen , He Su , A. Egrinya Eneji , Yu-hai Guo , Xue-hui Dong

2014, 6(2):145-151. DOI: 10.1016/S1674-6384(14)60023-X

[Abstract](918) [HTML](0) [PDF 328.27 K](2116)

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Objective To search for the potential medicinal plant part of Gentiana macrophylla based on changes of secondary metabolites and trace elements in the flowers of G. macrophylla. Methods HPLC was used to detect the changes of the active constituents (longanic acid, sweroside, gentiopicroside, and swertiamarin) and ICP-AES was used for mineral nutrients in G. macrophylla during flower development. And soluble sugar, starch, crude protein, hemicelluloses, cellulose, and lignin were determined. Results Biomass of flower in full bloom (D2) phase was considerable during flower development, in which the contents of longanic acid and gentiopicroside were at the highest levels with 2.65 and 2.88 times higher than those recorded in Chinese Pharmacopoeia 2010, sweroside and swertiamarin in the flowers were reaching 6.06 and 1.25 times higher than those in roots. Florescence is the most valuable stage during flower development. The concentration of Fe, Mg, K, P, and B was higher in the flowers than that in roots. The accumulation of active constituents in the plant was influenced by the contents of metabolically linked carbon and nitrogen compounds. Conclusion The secondary metabolites, mineral nutrients, and physicochemical indicators are tightly regulated by flower organ development, D2 is an important stage for both biomass and extraction of active constituents such as longanic acid. The flowers of G. macrophylla could be used as a potential medicinal plant part for longanic acid at a high level.

13 Estimation of Nuclear DNA Content in Tannin-rich Medicinal Plant Cornus officinalis by Flow Cytometry

Bo Cao , Lin-lin Zhang , Cheng-ke Bai

2014, 6(2):152-158. DOI: 10.1016/S1674-6384(14)60024-1

[Abstract](767) [HTML](0) [PDF 341.99 K](2293)

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Objective The amount of nuclear DNA (C-value) is a key biodiversity character that provides strong unifying elements in revealing the phylogenetic regularity and relationship between genome size and functional traits for plant resource. The estimation of C-values could primarily extend our knowledge on the genetic background and genome diversity for medicinal plants, and thereby the variation of pharmacological constituents and phylogenetic mechanism of medicinal plant taxa will be revealed. However, a large number of medicinal plants (e.g. Cornus officinalis) typically contain a series of secondary metabolites, especially tannic acid, which would significantly affect the estimation of DNA content by flow cytometry (FCM). Methodological discussions and improvement need to be made to solve this problem. Methods Two isolation buffers LB01 and Otto 1 were selected to prepare nuclear suspension with additional treatments of pre-soaking and centrifugation combination of gradient centrifugal force and duration. The best isolation and estimation methods were determined by FCM measurement in C. officinalis. Results The dry leaves were pre-soaked in Otto I buffer for 15 min and the Otto I nuclear suspension was centrifugated at 1.0×103 g for 2 min. The results showed that debris and nuclei were better separated and the scatterplots of good quality were obtained with low coefficient of variation (CV). Contrarily, the nuclear DNA content of C. officinalis could not be accurately estimated for nuclei extracted by LB01 buffer. Finally, 2C-value and genome size of C. officinalis were first estimated as 5.92 pg and 2893 Mbp, respectively. Conclusion The new methods proposed here are able to accurately estimate DNA content of C. officinalis, which provides valuable references for the estimation of genome size in other tannin-rich medicinal plants.

14 Saponins from Roots of Panax notoginseng

Li-feng Han , Kaunda Joseph Sakah , Li-li Liu , Agyemang Kojo , Tao Wang , Yi Zhang

2014, 6(2):159-163. DOI: 10.1016/S1674-6384(14)60025-3

[Abstract](1294) [HTML](0) [PDF 287.20 K](2350)

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Objective To study the chemical constituents in the dried roots of Panax notoginseng. Methods The constituents were isolated and purified with chromatographic methods. Their structures were elucidated by spectroscopic methods (1D, 2D NMR, UV, IR, [a]D, and HRESI-TOF-MS) and chemical analyses. Results Twenty saponins including 20(S)-ginsenoside Rh1 (1), 6-O-b-D-(6′-acetyl)-glucopyranosyl-24-ene-dammar-3b, 6a,12b,20S-tetraol (2), ginseno- side Rf (3), notoginsenoside R2 (4), ginsenoside Rg2 (5), ginsenoside Rg1 (6), notoginsenoside Rt (7), koryoginsenoside R1 (8), 6-O-(b-D- glucopyranosyl)-20-O-(b-D-xylopyranosyl)-3b,6α,12b,20(S)-tetrahydroxy-dammar- 24-ene (9), pseudoginsenoside Rt3 (10), notoginsenoside R1 (11), ginsenoside Re (12), notoginsenoside N (13), ginsenoside F1 (14), ginsenoside U (15), ginsenoside Rk3 (16), 3b,12b-dihydroxydammar-(E)-20(22),24-diene-6-O-b-D-xylopyranosyl-(1→2)-b-D- glucopyranoside (17), ginsenoside Rh4 (18), pseudoginsenoside Rt5 (19), and vinaginsenoside R22 (20) were obtained. Conclusion Compounds 2, 19, and 20 are isolated from this species for the first time. The 1H-NMR data of compound 19 and 1H-NMR and 13C-NMR data of compound 20 are first reported. Meanwhile, the NMR data of b-D-xylopyranosyl group in compound 9 is corrected.

15 Diacylglycerol Compounds from Barks of Betula platyphylla with Inhibitory Activity against Acyltransferase

Nan Zhang , Na Li , Ya-nan Sun , Jia-lin Li , Shan-shan Xing , Zhen-dong Tuo , Long Cui

2014, 6(2):164-167. DOI: 10.1016/S1674-6384(14)60026-5

[Abstract](764) [HTML](0) [PDF 343.63 K](2116)

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Objective To identify the active compounds from the barks of Betula platyphylla for inhibitory on diacylglycerol acyltransferase (DGAT1). Methods Bioassay-guided fractionation resulted in the isolation of DGAT1 inhibitory activity of lupane triterpenes. Results Ten compounds were identified as lupenone (1), lupeol (2), betulinic acid (3), betulinaldehyde (4), betulin (5), 3-deoxybetulonic acid (6), glochidonol (7), lup-20/29- ene-1β/3β-diol (8), 3α-hydroxy-lup-20(29)-en-23,28-dioic acid (9), and 3α,11α- dihydroxy-23-oxo-lup-20(29)-en-28-oic acid (10). Compounds 3-6, 9, and 10 inhibited DGAT1 with IC50 values ranging from (11.2 ± 0.3) to (38.6 ± 1.2) μmol/L. Conclusion Compounds 6, 9, and 10 are first isolated from the barks of B. platyphylla., and compounds 3-6, 9, and 10 from the barks of B. platyphylla are responsible for the inhibition on DGAT1.

16 A New Furan Flavonol Glycoside from Epimedium koreanum

Jin-dong Zhong , Cheng Zhou , Hong-mei Li , Rong-tao Li

2014, 6(2):168-170. DOI: 10.1016/S1674-6384(14)60027-7

[Abstract](771) [HTML](0) [PDF 318.80 K](1713)

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Objective To study the chemical constituents of Epimedium koreanum. Methods Separation was carried out through silica gel and Sephadex LH-20 column chromatography and HPLC method. The chemical structures were elucidated by spectroscopic method including 1D-NMR and 2D-NMR. Results A new furanflavonol glycoside (1) was isolated and identified. Conclusion Compound 1 is a new furanflavonol glycoside, and its structure is corroborated as 5,4′-dihydroxyfurano [2″,3″:7,8] flavonol 3-O-α-L-rhamnoside.

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