Botanical Studies (2010) 51: 317-324.
BIOCHEMISTRY
Antihypertensive activities of extracts from tissue
cultures of Vitis thunbergii var. taiwaniana
Chun-Yao HUANG1, Chi-Luan WEN2'3, Yeh-Lin LU4, Yin-Shiou LIN4, Lih-Geeng CHEN5,
and Wen-Chi HOu6,7 *
1Department of Internal Medicine, Taipei Medical University and Hospital, Taipei, Taiwan
2Taiwan Seed Improvement and Propagation Station, Council of Agriculture, Taichung, Taiwan
3 Graduate Institute of Chinese Pharmaceutical Science, China Medical University, Taichung, Taiwan
4School of Pharmacy, Taipei Medical University, Taipei, Taiwan
5Graduate Institute of Biomedical and Biopharmaceutical Sciences, National Chiayi University, Chiayi, Taiwan
6Traditional Herbal Medicine Research Center, Taipei Medical University Hospital, Taipei, Taiwan
7Graduate Institute of Pharmacognosy, Taipei Medical University, Taipei, Taiwan
(Received november 18, 2009; Accepted January 20, 2010)
ABSTRACT. Vitis thunbergii var. taiwaniana (VTT), the wild grape, is an endemic plant in Taiwan which has long been used as a folk medicine. Two ethanolic extracts (EE), including shoots (S) and root parts (R), from each plant growth regulator-treated VTT tissue culture (TC) plantlets, including TC1 (treated with IAA), TC2 (treated with IBA), TC3 (as the control), and TC4 (treated with NAA), were used to test the angiotensin converting enzyme (ACE) inhibitory activities. It was found that EE of R and S parts from four VTT-TC plantlets exhibited ACE inhibitory activities and the TC-R had better ones. The EE of whole plantlet ex­hibited dose-dependently ACE inhibitory activities (expressed as IC50), and the orders were TC1 (98.67 μg/ mL) > TC3 (99.04 μg/mL) > TC4 (102.46 μg/mL) > TC2 (132.05 μg/mL). The hot water extracts (HWE) of TC1-A and HWE-TC1-B exhibited dose-dependently ACE inhibitory activities and the IC50were 29.51 and 32.79 μg/mL, respectively. For short-term antihypertensive activity of EE-TC1-R in vi~vo, the spontaneously hypertensive rats (SHRs) were fed by a single oral adminisration (20 mg/Kg of SHR), and the changes of systolic blood pressure (SBP) and diastolic blood pressure (DBP) during 24-h were measured. It was found that EE showed antihypertensive activities and the higherst lowering effects was reached at 4th-h after being orally administered and the reductions of SBP and DBP were 16.9 and 17.7 mmHg, respectively. It can also be noted that the SBP reduction could last over 24-h. For long-term antihypertensive activity of EE-TC1-R in vivo, SHRs were fed orally once a day for four weeks (30 mg/Kg of SHR), and the changes of systolic blood pressure (SBP) and diastolic blood pressure (DBP) were measured weekly. It was found that the reductions of SBP, but not DBP, were 12.45, 17.13, and 10.05 mmHg, respectively, at the 2nd, 3rd, and 4th-week and showed significantly different (P<0.05 or P<0.01) compared to the blank. The ampleopsin C and (+)-s-viniferin were identified in the EE-TC1-R. This will be
beneficial to develope the extracts of VTT as functional foods for blood pressure regulation.
Keywords: Blood pressure; Plantlet of tissue culture (TC); Spontaneously hypertensive rats (SHRs); Vitis thunbergii var. taiwaniana (VTT).
introduction
A number of risk factors are associated with stroke, including age, gender, elevated cholesterol, smoking, alcohol, excessive weight, race, family history, and hypertension (Mark and Davis, 2000). Although some of these risk factors cannot be modified, one factor that can be controlled and has the greatest impact on the etiology of stroke is high blood pressure (Dunbabin, 1992).
*Corresponding author: E-mail: wchou@tmu.edu.tw; Fax: +886-2-2378-0134.
Several classes of pharmacological agents have been
used in the treatment of hypertension. One class of anti-
hypertensive drugs known as angiotensin I converting
enzyme (ACE) inhibitors (i.e. a peptidase inhibitor) are
associated with a low rate of adverse side-effects and are
the preferred class of anti-hypertensive agents for treating
patients with concurrent secondary diseases (Fotherby
and Panayiotou, 1999). ACE (EC 3.4.15.1) is a dipeptide-
liberating exopeptidase which has been classically
associated with the renin-angiotensin system regulating
peripheral blood pressure (Mullally et al., 1996). The
potent ACE inhibitors were frequently derived from food
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proteins (Ariyoshi, 1993; Hsu et al., 2002). However, pomegranate juice (Aviram and Dornfeld, 2001), flavan-3-ols and procyanidins (Actis-Goretta et al., 2003), myricetin galloylglycosides (Lee et al., 2006), small molecules of methanol-soluble, p-elimination products from preparations of alginic acid hydroxamate (Liu et al., 2007), geraniin (Lin et al., 2008), and tannins (Liu et al., 2003) were also reported to have ACE inhibitory activity. Fujita et al. (2000) found that the octapeptides of FFGRCVSP (IC50=0.4 μM) and ERKIKVYL (IC50=1.2 μM) were potent ACE inhibitors, but none of them were effective in animal models to reduce the blood pressure of SHRs. Sato et al. (2002) pointed out that three dipeptides, including AW (IC50=18.8 μM), VW (IC50=3.3 μM), and LW (IC50=23.6 μM), were potential ACE inhibitory peptides. However, none of them were able to effectively reduce the blood pressure of SHRs in animal models. Clearly, ACE inhibitory candidates in vitro might not have the antihypertensive effects on SHR in vivo.
Vitis thunbergii var. taiwaniana (VTT), as a wild grape, belonged to the Vitaceae family and Vitis genus, is an endemic plant in Taiwan which has long been used as folk medicines for treatments of hepatitis, jaundice, diarrhea, and arthritis (Chiu and Chang, 1995). The Endemic Species Research Institute, Council of Agriculture has cataloged VTT as an original medicinal plant in Taiwan. The active components from Vitis genus were reported to be oligostilbenes (Li et al., 1996; Teguo et al., 1998; Huang et al., 2001; Huang et al., 2005; Chen and Wang, 2009) and polyphenols (Dou et al., 2003). The methanolic root extracts of
VTT were reported to have anti-methicillin-resistant Staphylococcus aureus activities (Peng et al., 2008). In this report, ethanolic extracts (EE) and hot water extracts (HWE) of plant growth regulatortreated VTT tissue culture (TC) plantlets were used to examine the ACE inhibitory activities in vitro. The EE of VTT-TC were then to investigate the short-term and longterm antihypertensive activity by oral administrations of TC1-R to SHR in vivo. These results showed that it will be beneficial to develope the extracts of VTT as functional foods for blood pressure regulation.
materials and methods
Materials
ACE (I unit, rabbit lung) was purchased from Fluka Chemie GmbH (Switzerland); N-(3-[2-furyl] acryloyl)-Phe-Gly-Gly (FAPGG), and other chemicals and reagents were from Sigma Chemical Co. (St. Louis, MO, USA).
Plants
Four VTT-TC plantlets, including TC1, treated with
IAA (indole-3-acetic acid); TC2, treated with IBA (indole-3-butyric acid); TC3, as the control, without plant growth regulator treatment; and TC4, treated with NAA (naphthaleneacetic acid), which were provided by Dr. Wen, Chi-Luan (Taiwan Seed Improvement and Propagation
Station, Council of Agriculture, Taichung, Taiwan), were used in this study. The VTT plantlet was identified by Dr. Hsu, Tsai-Wen (Endemic Species Research Institute, Nantou, Taiwan). The photographs of whole TC plantlets were shown in Figure 1.
Preparation of ethanolic extracts and hot water extracts from VTT-TC
After being washed with distilled water thrice to remove the culture medium, the whole TC plantlets with or without being freshly cut into shoot (S) and root (R) portions. For ethanolic extracts (EE), 1,000 mL of absolute ethanol was added into whole TC plantlets or S portions (wet weight), or 500 mL of absolute ethanol was added into R portions (wet weight) and then extracted at room temperature for one week. After being filtered, the residue was extracted with ethanol once. The filtrates were collected and concentrated as EE extracts. For hot water extracts (HWE), the whole plantlets of VTT-TC1 were air-dried at 37°C oven and cut into pieces and placed into tea bags and then were sealed. The VTT tea bags were extracted twice by 100°C hot water in the ratio of 1/10 (w/v) for 30 min. After being filtered, the filtrates were collected and then lyophilized as the HWE-TC1. There were two methods to culture VTT-TC1 (treated with IAA plant regulators) plantlets on the same 90-days for HWE-TC1 preparations (including HWE-TC1-A and replacing the old culture medium after 30-days with a new one, and then for another 60 days; TC1-B represented the culture method of directly adding the new culture medium onto the old one after 30-days, and then for another 60 days
.
Determination of the ACE inhibitory activity of different VTT-TC extracts by spectrophotometry
The ACE inhibitory activity was measured according to the method of Holmquist et al. (1979) with some modifications. Twenty μL (20 mU) of commercial ACE (1 U/mL, from rabbit lung) were mixed with 200 (g of EE from TC1-R, TC1-L, TC2-R, TC2-L, TC3-R, TC3-L, TC4-R, and TC4-L, or different amounts of whole plantlets of EE (50-250 μg) from TC1, TC2, TC3, and TC4, or different amounts of HWE-TC1-A and HWE-TC1-B (16.39, 32.79, 49.18, and 65.57 μg/mL), and then 1 mL of 0.5 mM FAPGG [dissolved in 50 mM Tris-HCl buffer (pH 7.5) containing 0.3 M NaCl] was added. The decreased absorbance at 345 nm (ΔAsample) was recorded within a 1.5-min span at room temperature and expressed as ΔAsample/ min. The extracted solvent, ethanol or distilled water, respectively, was used in blank experiments and expressed as AAblank/min. The ACE inhibition (%) was calculated as follows: [1 - (ΔAsample/min ÷ ΔAblank/min)] x 100%. Means of triplicates were determined. The 50% inhibition (IC50) of ACE activity was calculated as the concentrations of samples that inhibited 50% of ACE activity under these conditions.
HUANG et al. ― Antihypertensive activities of VTT extracts
319
Figure 1. The photographs of whole plantlet of four typers of tissue culture (TC) of Vitis thunbergii var. taiwaniana as followed: TC1, IAA (indole-3-acetic acid); TC2, IBA (indole-3-butyric acid); TC3, the control; and TC4, NAA (naphthaleneacetic acid).
Antihypertensive effects of EE-TC1-R on SHR
The effects of orally-administered EE-TC1-R by feeding tube (2.0 x 80 mm) on the blood pressure of SHR were determined (Lin et al., 2006; Liu et al., 2007; Lin et al., 2008; Liu et al., 2009a, b). All animal experimental procedures followed published guidelines (National Science Council, 1994) and reviewed and approved by the Institutional Animal Care and Use Committee of Taipei Medical University (LAC-95-0076). The male SHRs (8 weeks of age, National Laboratory Animal Center, Taipei) were housed individually in steel cages kept at 24°C with a 12-h light-dark cycle and had free acess to a standard mouse/rat chow (ProlabR RMH2500, 5P14 Diet, PMI Nutrition International Brentwood, MO) and water. SHRs were randomly divided into control and sample treatments for blood pressure determinations (six rats per group). For a short-term antihypertensive experiment, 0.5-mL of water-dissolved EE-TC1-R was orally administered to SHR (20 mg/Kg of SHR) once, and tail blood pressure was measured four times at each desired time over 24 h using an indirect blood pressure meter (BP-98A, Softron Co. Ltd. Tokyo, Japan) for systolic blood pressure (SBP) and diastolic blood pressure (DBP) measurements. For long-term antihypertensive effects, the EE-
TC1-R was orally administered to SHR once a day for 4-weeks (30
mg/Kg of SHR) and the blood pressure was measured once every week before each oral administration. Before each blood pressure measurement, SHRs were warmed for 10 min in a 39°C thermostated box. The 0.5-mL distilled water was used for a blank experiment and the blood pressure was measured after oral administration of distilled water. Means of triplicates were recorded. The measured blood pressure values were collected and averaged from six rats as termed BPblank. The measured blood pressure values of each rat after being administered sample orally were collected and averaged as termed the BPsample. The six values calculated from BPsample - BPblank are averaged and then indicated as lowering effects in blood pressure changes (ΔBP) in sample at the same treatment time after oral administration (such as 2, 4, 6, and 24-h) for short-term antihypertensive activity. Means ± SD of triplicates were measured. Student's t-test was used for comparisons between blank and sample treatment at the same time interval when P< 0.05 (*) or P< 0.01 (**).
HPLC chromatogram of EE-TC1-R of Vitis
thunbergii var. taiwaniana
The HPLC chromatogram of EE-TC1-R was performed by XTerra MS C18 HPLC column (3.5 (m, 2.1x150 mm). The mobile phase was mixed in gradients with 95%
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Solvent A and 5% Solvent B (0 min) to 70% Solvent A and 30% Solvent B (50 min). Solvent A: 0.1% TFA, and Solvent B: acetonitrile. The detector was set at UV 280 nm. The identified component, ampelopsin C (35.8 min) and (+)-s-viniferin (38.6 min), were compared to each isolated pure compound and electrospray ionization mass spectra (ESI-MS) (Chen and Wang, 2009).

results and discussion
Vitis thunbergii var. taiwaniana (VTT), as a wild grape, belonged to the Vitaceae family and Vitis genus, is an endemic plant in Taiwan which has long been used as folk medicines for treatments of hepatitis, jaundice, diarrhea, and arthritis (Chiu and Chang, 1995). The Endemic Species Research Institute, Council of Agriculture has cataloged VTT as an original medicinal plant in Taiwan. Therefore, the evidence-based biological functions of VTT might be main criteria to develop as functional foods.
The screening of ACE inhibitory activities in vitro might be reasonable for further antihypertensive activities in animal models. Figure 2 showed the 200 (g of EE extracts from R and S portions of four VTT-TC on 20 mU ACE. It was clear that the FAPGG was hydrolyzed by ACE to produce FAP and resulted in the reduction of absorbance at A345 nm (Holmquist et al., 1979). It could be found that the blank (ACE and FAPGG) exhibited the highest reduction of AAblank/min and the R portion had better effects than S portion to inhibit the reductions in AAsample/min (Figure 2A). It was calculated for ACE inhibition under the same 200 (g of EE extracts (Figure 2B), the orders of each ACE inhibition of TC-VTT were TC3-R (59.22%) ~ TC1-R (58.25%) ~ TC2-R (55.83%) > TC4-R (48.06%) >> TC1-S (23.79%) ~ TC3-S (22.82%) > TC4-S (16.02%) > TC2-S (2.91%). It was preliminarily found that the IAA-treated TC-VTT (TC1) had closely ACE inhibitory activities to the control TC-VTT (TC3) and better than other plant growth regulator-treated VTT-TC. Therefore, the dose effects of EE from the whole plantlets of VTT-TC were investigated and the results were shown in Figure 3. It was clear that the dose-dependent ACE inhibition was found and the IC50 of ACE inhibitory respectively, for TC1, TC2, TC3, and TC4. The IAA-treated plantlet (TC1) had higher ACE inhibitory activities compared to the control plantlet (TC3) used in this study. It was the first report that EE of VTT-TC exhibited ACE inhibitory activities.
For contamination prevention during medium replacements for TC, two cultured methods for TC1 were used. TC1-A represented the cultured method of replacing the old culture medium after 30-days with a new one, and then for another 60 days; TC1-B represented the culture method of directly adding the new culture medium onto the old one after 30-days, and then for another 60 days. Both whole TC plantlets were washed, air-dried at 37°C and then cut into pieces and placed and simulated as tea bags. Figure 4 showed the effects of different
Figure 2. Effects of ethanolic extracts (200 (g) of two portions (S, shoots; R, roots) of four tissue culture (TC) of Vitis thunbergii var. taiwaniana on 20 mU ACE inhibition by continuous spectrophotometric methods. (A) The decreased absorbance at 345 nm ((ΔAsample-(ΔAblank) were recorded during 1.5 min at room temperature and expressed as (ΔA/min. (B) The ACE inhibition (%) was calculated according to the equation of [1-((ΔA sample /min +(ΔAblank/min) ] x 100%.
Figure 3. Effects of different concentrations of ethanolic extracts of whole plantlets of four tissue culture (TC) of Vitis thunbergii var. taiwamiama on 20 mU ACE inhibition by continuous spectrophotometric methods. The ACE inhibition (%) was calculated according to the equation of [1-((ΔAsample / min + (ΔAblank/min) ] x 100%. The 50% inhibition (IC50) of ACE activity was calculated as the concentrations of samples that inhibited 50% of ACE activity under these conditions.
HUANG et al. ― Antihypertensive activities of VTT extracts
321
concentrations of HWE-TC1-A and HWE-TC1-B on ACE
inhibitions. It was found that HWE from whole plantlets of VTT-TC exhibited dose-depently ACE inhibitory activities and the IC50of ACE inhibitory activity was 29.51 and 32.79 μg/mL, respectively, for HWE-TC1-A and HWE-TC1-B. It was found that HWE of two cultured methods from whole plantlets of TC1 for ACE inhibition were closely and three-folds better than that of EE (Figure 3).
Some reports had pointed that ACE inhibitory candidates in vitro might not have the antihypertensive effects on SHR in vivo (Fujita et al., 2000; Sato et al., 2002). Therefore, the EE-TC1-R was orally administered to SHR (20 mg/Kg of SHR) once, and the ABP was measured four times at different time intervals within 24 h for short-term effects (Table 1). It was found that
EE-TC1-R at dose of 20 mg/Kg of SHR could lower the SBP and DBP of SHR within 24-h. For changes of SBP (ASBP), there were 14.1, 16.3, 9.1, and 10.9 mmHg reductions, respectively, for the 2nd, 4th, 6th, and 24th-h after being orally administered. For changes of DBP (ΔDBP), there were 11.1, 17.7, 11.9, and 6.7 mmHg reductions, respectively, for the 2nd, 4th, 6th, and 24th-h after being orally administered. It was noted that the SBP reductions could over 24-h by a single oral administration of EE-TC1-R. Figure 5 showed the effects of EE-TC1-R (30 mg/Kg of SHR) on the changes of SBP and DBP of SHR
Table 1. Effects of ethanolic root extracts of tissue culture of Vitis thunbergii var. taiwaniana (TC1-R, treated with IAA plant growth regulators) on the changes of blood pressure of sponta­neously hypertensive rats (20 mg/Kg of SHR) during 24-h by a single oral administration.
Time after TC1-R extracts (20 mg/Kg of SHR)
treatment (H)
ASBP (mmHg)*
ADBP (mmHg)*
2
-14.1 ± 2.9
-11.1 ± 4.6
4
-16.3 ± 6.8
-17.7 ± 5.7
6
-9.1 ± 5.3
-11.9 ± 4.4
24
-10.9 ± 7.0
-6.7 ± 6.0
*The lowering effects in blood pressure changes (ΔBP) were calculated as BPsample - BPblank at the same treatment time after oral administration (n=6).
Concentration (jjg/mL)
Figure 4. Effects of hot-water extracts (HWE) of dried whole plantlets of tissue culture of Vitis thunbergii var. taiwaniana (TC1, treated with IAA plant regulators) on 20 mU ACE inhibition by continuous spectrophotometric methods. TC1-A represented the cultured method of replacing the old culture medium after 30-days with new culture medium, and then for another 60 days; TC1-B represented the culture method of directly adding the new culture medium onto the old culture medium after 30-days, and then for another 60 days. The dried whole plant was extracted twice by 100°C hot water in the ratio of 1/10 (w/v) for 30 min. After being filtered, the filtrates were collected and then lyophilized as HWE. The ACE inhibition (%) was calculated according to the equation of [1- (ΔAsample/ min + ΔAblank/min) ] x 100%. The 50% inhibition (IC50) of ACE activity was calculated as the concentrations of samples that inhibited 50% of ACE activity under these conditions.
Figure 5. Effects of ethanolic extracts of root portions of tissue culture (TC) of Vitis thunbergii var. taiwaniana (TC-1R, treated with IAA plant regulators) on systolic blood pressure and diastolic blood pressure of spontaneously hypertensive rats (SHR, n=6) by oral administration (30 mg/Kg of SHR, dissolved in 0.5-mL distilled water) once a day for four-weeks and 0.5-mL distilled water was used for a control experiment. A difference was considered statistically significant between control and
treated group when P< 0.05 (*) or P< 0.01 (**).
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Figure 6. HPLC chromatogram of the ethanolic extracts of root parts of tissue culture of Vitis thunbergii var. taiwaniana (EE-TC1-R). The main components of ampleopsin C (35.8 min) and (+)-s-viniferin (38.6 min) were identified by pure compound and ESI-MS. Column, XTerra MS C18, 3.5 (m (2.1x150 mm); mobile phase, 0 min: 95% Solvent A and 5% Solvent B; 50 min, 70% Solvent A and 30% Solvent B in gradients. Solvent A: 0.1% TFA, and Solvent B: acetonitrile. Detector, UV 280 nm.
over four weeks. It was found that the reductions of SBP, but not DBP, were 12.45, 17.13, and 10.05 mmHg, respectively, at the 2nd, 3rd, and 4th-week and showed significantly different (P<0.05 or P<0.01) compared to the blank. It was the first report that EE-TC1-R exhibited ACE inhibitory and antihypertensive activities.
Figure 6 showed the HPLC chromatogram of EE-TC1-R. It was found that the main components of ampleopsin C (35.8 min) and (+)-s-viniferin (38.6 min) were identified by each pure compound and ESI-MS (Chen and Wang, 2009). Calculation from area percentage, ampleopsin C and (+)-s-viniferin was accounted for 18.24% and 12.98%, respectively, in EE-TC1-R extracts.
In literatures, (+)-hopeaphenol, isohopeaphenol, vitisin A, (+)-vitisifuran A, and heyneanol A isolated from Vitis amurensis showed inhibitory activities against leukotriene B4 biosynthesis (Huang et al., 2001). Vitisin A is one component of Vitis spp. that exerts anti-inflammatory activity by inhibiting influenza A virus-induced cytokine production (Huang et al., 2008). Peng et al. (2008) demonstrated the antimicrobial activity of heyneanol A, a component of VTT root, against MRSA pathogens. The vitisin A exhibited inhibitory activity against adipoyte differentiations (Kim et al., 2008). It might be possible that ampleopsin C, (+)-s-viniferin and others might contribute the antiphypertensive activity in vivo and needed further investigations.
In conclusion, EE and HWE of VTT-TC exhibited ACE
inhibitory and/or antihypertensive activities against SHRs in this report. The ampleopsin C and (+)-s-viniferin were identified in the EE-TC1-R. For commercial uses, effects of extracts from the field cultivations of mature plants from a fixed TC plantlet and pure compound isolation for mechanism elucidation will be performed recently. These results showed that it will be beneficial to develope the extracts of VTT as functional foods for blood pressure regulation.
Acknowledgments. The authors want to thank the financial supports (96AS-1.2.1-ST-a1(20) and 97AS-1.2.1-ST-a1(26)) from Council of Agriculture, Republic of China.
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Botanical Studies, Vol. 51, 2010
小葉葡萄組織培養苗之抽取物降血壓功效之研究
黃群耀1 文紀鑾2,3 呂岳霖4 林音秀4 陳立耿5 侯文琪6,7
1臺北醫學大學醫學系及附設醫院心臓內科
2農委會種苗改良繁殖場
3中國醫藥大學中國藥學研究所
4臺北醫學大學藥學系
5嘉義大學生物醫藥科學研究所
6臺北醫學大學附設醫院中草藥研究中心
7臺北醫學大學生藥學研究所
小葉葡萄(Vitis thunbergii var. taiwaniana)爲台灣特有之變種。本硏究以不同植物生長素進行90天
培養之小葉葡萄組織培養苗爲材料（TC1, IAA; TC2, IBA; TC3,對照組；TC4, NAA)'發現其莖葉部（S)
與根部(R)之酒精抽取物具有抑制血管收縮素轉化酶抑制活性，但以根部效果較佳。四種組織培養之小
葉葡萄苗之酒精抽取物50%抑制血管收縮素轉化酶活性(IC50)分別爲TC1 (98.67 μg/mL) > TC3 (99.04
μg/mL) > TC4 (102.46 μg/mL) > TC2 (132.05 μg/mL)。以TC1之熱水抽取物爲材料'發現30天後倒
掉舊培養基，加入新培養基培養60天（HWE-TC1-A)，或是添加新培養基於舊培養基中共同培養60天
(HWE-TC1-B)，皆能有效抑制血管收縮素轉化酶活性，IC50分別爲29.51 μg/mL (HWE-TC1-A)與32.79
μg/mL (HWE-TC1-B)。先以組織培養苗根部酒精抽取物(TC1-R)進行高血壓鼠餵食一次（20 mg/Kg),
觀察24小時血壓變化之試驗。結果顯示，組織培養苗根部醇類抽取物(TC1-R)於第四小時達到最低的
血壓，其收縮壓與舒張壓分別降低16.9毫米汞柱與17.7毫米汞柱，24小時後收縮壓還有11毫米汞柱
的降幅。以組織培養苗根部酒精抽取物(TC1-R)進行高血壓鼠每天餵食一次(30 mg/Kg)'觀察四週血壓
變化。結果顯示，組織培養苗根部酒精抽取物（TC1-R)，餵食後的收縮壓於第二、三與四週，分別下降
12.45, 17.13與10.5毫米汞柱，並與對照組呈現顯著性差異(P<0.05或P<0.01)；但舒張壓並沒有明顯差
異。小葉葡萄酒精抽取物與熱水抽取物未來可以開發爲調節血壓之保健品。
關鍵詞：小葉葡萄；組織培養苗；高血壓鼠；血壓。