Studies On The Sedative Effect Of Cistanche Deserticola

for more information:Ali.ma@wecistanche.com

Ming-Chin Lu

Department of Chinese Diagnosis, China Medical College, 91 Hsieh Shih Road, Taichung, Taiwan, ROC

Abstract

In this study, we investigated the sedative effect of Cistanche deserticola Ma. (CD) on hexobarbital-induced sleeping time in mice and spontaneous motor activity by using an automated activity meter in rats. It was found that crude extract of CD (Cistanche deserticola)could prolong the hexobarbital-induced sleeping time and reduce spontaneous motor activity, including horizontal activity, ambulatory time, and total distance. Then the water fraction of CD (Cistanche deserticola) extract could prolong the hexobarbital-induced sleeping time and reduce the spontaneous motor activity more than that of the other fraction of CD (Cistanche deserticola) extract in rats. These results suggest that CD ethanol extract and its water fraction possessed the sedative effect. © 1998 Elsevier Science Ireland Ltd.

Keywords: Cistanche deserticola; Sedation; Automated activity meter; Spontaneous motor activity

Cistanche deserticola has a sedative effect

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1. Introduction

Cistanche deserticola Ma. (Orobanchaceae) (CD) is Chinese traditional medicine, widely used in the treatment of insomnia, pain, fatigue, and amnesia (Lee, 1986). In modern pharmacological studies, ethanol extract of CD (Cistanche deserticola) is found to have anti-aging effects (Lee et al., 1990) and enhance immune functions (Chin and Su, 1993). Water extract of CD extract increases DNA synthesis and SOD activity (Chang et al., 1995) and excludes free radicals (Hsu et al., 1995). However, the sedative effect of CD has never been studied. The sedative effect of drugs can be evaluated by measuring spontaneous motor activity in rats (Carpenedo et al., 1994; File and Fernandes, 1994; Hsieh et al., 1991). So, the purpose of the present study was intended to evaluate the sedative effect of CD by measuring hexobarbital-induced sleeping time in mice and locomotor behavior, including horizontal activity, ambulatory time, and total distance in rats, then separate the CD extract with different solvents to search for its active ingredients.

2. Materials and methods

2.1. Preparation of CD (Cistanche deserticola)

Rhizomes of Cistanche deserticola Ma. (Orobanchaceae) were purchased from the Taiwan market and were authenticated by Dr. YungHsun Chang, China Medical College, Taichung. A specimen has been deposited in the Department of Botany, China Medical College (identification no. 15).

Rhizomes of CD (Cistanche deserticola) (2.5 kg) were chopped and extracted with 50% ethanol (3×3.5 l) by maceration (3 days) and the extract was reduced to dryness with a vacuum rotary evaporator. A yield of 840 g (33.6%) was obtained. The extract was dissolved in water and the solution was extracted successively with ethyl acetate and n-butanol (Hsu et al., 1994) to give the fractions in yields of 24.9 g (3.0%; FrEtOAc,) and 81.2 g (9.7%; FrBuOH,), respectively. The aqueous fraction yield was 242.3 g (28.8%; FrH2O,). The separation steps are shown in Fig. 1.

Fig. 1. Separation of rhizoma of Cistanche deserticola (CD).

2.2. Animals

Male Sprague-Dawley rats, weighing between 200–250 g were used for the study of motor activity. Male ICR mice, weighing between 20–25 g, were used for the study of the hypnotic effect. All animals were maintained in groups of five per cage at a temperature-control of 21–25°C and humidity of 5595%. The animals were housed in a metal-wire mesh cage for at least 7 days before each experiment. A standard pellet diet and tap water were provided ad libitum.

2.3. Hexobarbital-induced sleeping in mice

In the experimental group, each fraction of CD (Cistanche deserticola) extracts was administered orally at a dose of 0.1, 0.3, 1.0 g/kg body weight to different groups of mice. One hour later, hexobarbital (100 mg/kg, i.p.) was given to induce sleep. The interval between loss and recovery of righting reflex was used as the index of the hypnotic effect (Fujimori, 1965). In the control group, normal saline was used instead.

2.4. Measurement of spontaneous motor acti6ity in rats

The Opto-Varimex animal activity meter was used (Columbus Instruments, USA) for these measurements. The apparatus consisted of an acrylic activity monitor cage (43.2×44.4×30.5 cm) which was surrounded by horizontal and vertical sensors that are non-detectable by rats. The apparatus also consisted of fifteen infrared beams, spaced 2.65 cm apart on each axis. Two infrared beams were set as the sensitivity of monitors. The cage was enclosed in a ventilated cabinet. Measurements were performed between 09:00– 17:00 h.

Interruption of photocell beams was collected by a microcomputer and the following variables were recorded: (1) Horizontal activity (HA): total number of interruptions of horizontal beams; (2) ambulatory time (AT): time spent in ambulatory activity as measured by the interruption of different beams; and (3) total distance (TD): the distance moved by the animal (cm) during a movement period, as measured by repeated interruption of the same beam or movement below two beams with a break of 1 s, include grooming, scratching and digging (Sanberg et al., 1985). The activity was recorded for 1 h, starting 5 min after inserting the animal into the test cage. Each rat was used only once and a total of six rats was used for each treatment. Each fraction of CD(Cistanche deserticola) extract was administered orally at a dose of 0.1, 0.3, 1.0 g/kg body weight to different groups of rats. One hour later, spontaneous motor activity was recorded. Control rats received vehicles only.

Cistanche deserticola has a sedative effect

2.5. Chemicals

Methanol, n-butanol, and ethyl acetate were HPLC grade, obtained from E. Merk.

2.6. Statistical analysis

All data were analyzed statistically using ANOVA followed by Duncan’s multiple range test to compare their differences. PB0.05 was required for statistical significance.

3. Results

3.1. Hexobarbital-induced sleep in mice

As shown in Table 1, ethanol extract of CD (Cistanche deserticola) at 0.1, 0.3, and 1.0 g/kg prolonged the hexobarbitalinduced sleeping time in mice (PB0.01). Between each fraction of CD (Cistanche deserticola) extract, only the water fraction of CD extract at 0.01, 0.03, and 0.3 g/kg could prolong the hexobarbital-induced sleeping time in mice (PB0.01).

3.2. Measurement of spontaneous motor acti6ity in rats

As shown in Fig. 2, ethanol extract of CD (Cistanche deserticola) at 0.1, 0.3, and 1.0 g/kg produced a significant decrease in all of the rat's spontaneous motor activities, including horizontal activity, ambulatory time, and total distance (PB0.01). Between each fraction of CD(Cistanche deserticola) extract, only the water fraction of CD (Cistanche deserticola) extract at 0.01, 0.03, and 0.3 g/kg could produce a significant decrease in all spontaneous motor activities in rats (PB0.01) (Fig. 3).

Fig. 2. Effect of ethanol extract of CD (Cistanche deserticola) on the changes in spontaneous motor activity in rats. Data are represented as mean9S.E.M. of six rats. ANOVA for repeated measures followed by Duncan’s multiple range test. ** PB0.01, as compared with the VEH group

Fig. 3. Effect of each fraction of CD (Cistanche deserticola) extracts on the changes in spontaneous motor activity in rats. Data are represented as mean9S.E.M. of six rats. ANOVA for repeated measures followed by Duncan’s multiple range test. ** PB0.01, as compared with the VEH group.

4. Discussion

It was found that ethanol extract of CD (Cistanche deserticola) could prolong the hexobarbital-induced sleeping time in mice. Then the water fraction of CD (Cistanche deserticola) extract could prolong the hexobarbital-induced sleeping time more than the other fraction of CD extract in mice. Furthermore, ethanol extracts of Cistanche deserticola Ma. could significantly decrease all spontaneous motor activities, including horizontal activity, ambulatory time and total distance in rats from the result shown in Fig. 2. The water fraction of CD (Cistanche deserticola) extract could produce a significant decrease in all spontaneous motor activities in rats among each fraction of CD (Cistanche deserticola) extract.

In recent phytochemical research, it was found that the water layer of CD (Cistanche deserticola) extracts involved cistanoside E-I, 8-epideoxylogenic acid, and geniposidic acid. These constituents possessed many physiologic actions and pharmacologic activity (Chin and Chang, 1994). Therefore, the sedative effect of the water layer of CD (Cistanche deserticola) extract might be the action of cistanoside, 8-epideoxylogenic acid, and geniposidic acid. The sedative constituents of the water layer of CD extract will be studied later.

In conclusion, CD (Cistanche deserticola) ethanol extract and its water fraction possessed the sedative effect.

Acknowledgments

I offer my sincere thanks to Dr. Ming-Tsuen Hsieh of the Institute of Chinese Pharmaceutical Sciences for his technical assistance.

Cistanche deserticola has a sedative effect

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