Antifatigue Activity Of Phenylethanoid-rich Extract From Cistanche Deserticola

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A phenylethanoid-rich extract (ECD) of Cistanche deserticola Y.C. Ma, a holoparasitic plant and a valuable traditional Chinese medicine, was evaluated for antifatigue activity in ICR mice. ECD (0.25, 0.50, 1.00 g/kg) was administered orally to mice for 3 weeks. The swimming time to exhaustion was longer in the treatment groups (0.50, 1.00 g/kg) than in the control group (p < 0.01). The serum creatine kinase, lactate dehydrogenase, and lactic acid levels were decreased significantly in the treatment groups compared with the control group, while the contents of hemoglobin and glucose were increased significantly. In conclusion, ECD appeared to enhance the swimming capacity of mice by decreasing muscle damage, delaying the accumulation of lactic acid, and improving energy storage. These results provide scientific evidence for the traditional Chinese medical practice of C. deserticola. Copyright © 2009 John Wiley & Sons, Ltd.

Keywords: Cistanche deserticola; antifatigue activity; phenylethanoid; biochemical parameters.

Antifatigue activity in Cistanche deserticola

INTRODUCTION

Cistanche deserticola (Orobanchaceae), a holoparasitic plant, can be found in the desert region of northwestern China. It is used as a traditional Chinese tonic drug for kidney deficiency, characterized by impotence, pain in the loins and knees, female sterility, and constipation due to dryness of the bowel in the senile. The plant is called ‘desert ginseng’ by local inhabitants because of its ginseng-like tonic effects. In fact, C. deserticola was frequently used in ancient prescriptions for the development of physical strength. Its efficacy in increasing physical stamina had been recorded in ancient Chinese medical books (Shang, 1993).

Pharmacological studies showed that Cistanche deserticola extracts possessed antinociceptive, antiinflammatory (Lin et al., 2002), and sedative effects (Lu, 1998). Phenylethanoids and polysaccharides, which are the major active constituents of this plant, were reported to have antioxidative (Xiong et al., 1996), hepatocyte protective effect (Xiong et al., 1998), NO radical-scavenging activity (Xiong et al., 2000), and immunological activity (Dong et al., 2007). However, few studies have focused on antifatigue activity. A previous study found that the polysaccharide-rich extract contributed little to the antifatigue activity of Cistanche deserticola (data not shown). It was hypothesized that phenylethanoid-rich extract (ECD), containing another major group of chemical constituents, may play an important role in its antifatigue activity. The purpose of this study was to evaluate the effect of ECD on the physical strength and endurance of mice during forced swimming experiments.

antifague acitivity in Cistanche deserticola

MATERIALS AND METHODS

Plant material. The fresh fleshy stems of C. deserticola were provided by the Yongning C. deserticola plantation (Ningxia, China). A voucher specimen was deposited in the Herbarium of the Institute of Medicinal Plant Development (Chinese Academy of Medical Science, China).

Preparation of the extracts. The air-dried plant material was powdered and extracted by percolation with 70% EtOH. The percolate was evaporated under reduced pressure and the residual fluid was filtered. The filtrate was concentrated and chromatographed on an AB-8 macroporous resin (The Chemical Plant of Nankai University, China) column using 50% and 95% EtOH in water as eluants. The 50% EtOH eluant was concentrated and dried under reduced pressure to obtain a phenylethanoid-rich fraction. The yield of the fraction was about 3.33% and the content of the phenylethanoids was 58.9%. Verbascoside and echinacoside were two major constituents in this fraction, with their contents being 5.6% and 33.3%, respectively.

Animals, groupings, and experiment design. Four-week-old male ICR mice were randomly divided into four groups. ECD at doses of 0.25, 0.50, 1.00 g/kg body weight was given to mice by intragastric administration, successive medication for 3 weeks, while the control group received the same volume of 0.9% saline. The mice were trained to swim for 20 min twice a week to customize to swim. After 3 weeks, the mice fasted overnight before forced swimming studies or blood sampling was performed. All experiments were carried out according to the National Institutes of Health Guide for Care and Use of Laboratory Animals and were approved by the Animal Ethics Committee of the Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences.

Weight-loaded swimming test. Fifty-six mice were used in this experiment as shown in Table 1. The test procedure was the same as described by Porsolt et al. (1977). Briefly, 30 min after the last intragastric administration, the mice were placed individually in a swimming pool
(65 × 45 × 40 cm) with 35 cm depth of water maintained at 25.5 ± 0.5 ºC. A tin wire (5% of body weight) was loaded on the tail root of the mouse. Exhaustion was determined by observing failure to swim and the swimming period was regarded as the time spent by the mouse floating in the water with struggling and making necessary movements until exhausting its strength and drowning. The mice were assessed to be exhausted when they failed to rise to the surface of the water to breathe within a 10 s period. The swimming time to exhaustion was used as the index of the forced swimming capacity.

Determination of tissue glycogen. After the weight-loaded swimming test, the liver and gastrocnemius muscle of the mouse were immediately dissected out, frozen in dry ice, and kept at -80 ºC until analysis of glycogen content was performed. The glycogen content was measured following the method described elsewhere (Chun and Yin, 1998).

Determination of blood biochemical parameters. With the purpose of collecting enough blood for assay, another 48 mice were used. Then 30 min after the last intragastric administration of ECD, the mice were forced to swim in the swimming pool (weight-unloaded) for 90 min. After anesthetization with ether, the blood was respectively collected in heparinized tubes and without anticoagulant tubes by extirpating the left eyeball. The whole blood samples were used to determine the hemoglobin and lactic acid according to the HiCN (hemoglobin cyanide) method (ASTM F 756-93, -86) and modified Barker-Summerson method (Pryce, 1969), respectively. Serum was prepared by centrifugation at 1000 × g, 4 ºC for 15 min and the levels of serum creatine kinase, lactate dehydrogenase, urea nitrogen, total protein, and glucose were determined by an automatic biochemical analyzer (Hitachi 7060, Hitachi, Ltd, Japan) with commercial kits (Biosino Biotechnology Co., Ltd, Beijing, China).

Statistics. The results were expressed as the mean ± SD. Comparisons between groups were made using Student's t-test.

antifatigue activity in cistanche deserticola

RESULTS AND DISCUSSION

Table 1 shows that swimming time was significantly longer in the ECD-administered groups (0.50, 1.00 g/kg). This result demonstrated that ECD possessed an antifatigue effect. In order to clarify its mechanism, the biochemical parameters related to fatigue were assessed in both ECD-treated and non-treated forced swimming mice.

Creatine kinase and lactate dehydrogenase are known to be accurate indicators of muscle damage (Burr et al., 1997; Coombes and McNaughton, 2000). A rise in the level of creatine kinase and lactate dehydrogenase in the blood indicates that muscle damage has occurred, or is occurring. Table 2 shows the lactate dehydrogenase and creatine kinase levels were decreased significantly in the ECD-administered groups (0.50 g/kg or 1.00 g/kg), indicating that ECD could decrease muscle damage during forced swimming.

Hemoglobin is commonly stated to be one of the major factors that can improve endurance capability by carrying oxygen to the tissues. The level of lactic acid, as a metabolic by-product of anaerobic glycolysis, has an inverse relationship to swimming time. It was observed that the hemoglobin level was increased significantly and lactic acid was decreased significantly in the ECD-administered groups (0.50, 1.00 g/kg).

Strikingly, little difference was found in the glycogen contents of the liver or muscle between the control and ECD-treated groups after the swimming test (Table 2). As is known, glycogenolysis predominates over gluconeogenesis during violent exercise. Thus, a slight rise in the amount of hepatic glycogen, together with the significant rise in the glucose content in the ECD-treated groups (0.50, 1.00 g/kg), suggested that ECD could improve the storage of hepatic glycogen.

In conclusion, ECD enhanced the swimming capacity of mice by decreasing muscle damage, delaying the accumulation of lactic acid, and improving energy storage. However, further study is still needed to elucidate the more exact mechanism of the effect of the ECD on fatigue and/or exercise durability.

Acknowledgments

This work was supported by the National Natural Science Foundation of China (No. 30672659) and the Ningxia Science and Technology Office (No. 05GG-10807). The authors are grateful to Dr. Elaine Waldron in Johns Hopkins University School of Medicine for her assistance in checking this manuscript.

antifatigue activity in cistanche deserticola

From: ' Antifatigue Activity of Phenylethanoid-rich Extract from Cistanche deserticola' by Run-Lan Cai et.al.

----PHYTOTHERAPY RESEARCH Phytother. Res. 24: 313–315 (2010) Published online 16 July 2009 in Wiley InterScience (www.interscience.wiley.com) DOI: 10.1002/ptr.2927

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