Effects of Jiawey Siwu on rat activity

American Journal of Chinese Medicine, Summer-Fall, 2001 by Zhi-Hong Jian, Chien-Chih Chen, Hsi-Yen Lu, Chih-Pang Chu, Yee-Yung Ng, Sue Yu

Abstract: Jiawey Siwu (JS) is a modified formula of Siwu with the addition of Ginseng Radix and Eucommiae Cortex. We studied the CNS effect of JS on pentobarbital-induced hypnosis in ICR mice after acute oral administration. We also investigated the motor activity and exercise performance effects of JS in SD rats after subchronic (12 days) and chronic (8 weeks) oral administration with an activity cage and rotarod, respectively. The duration of pentobarbital-induced hypnosis was significantly decreased in mice after acute oral administration of 0.21, 1.05 or 4.2 g/kg JS. The time of ambulatory activity and time on the rotarod significantly increased in rats after subchronic and chronic oral administration of 1.05 g/kg JS, respectively. These results indicate that JS potentially increased the excitability of the CNS in mice, as well as increasing motor activity and physical endurance in rats.

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Siwu is a Chinese herbal medicine composed of Angelicae Sinensis Radix, Rehmanniae Radix et Rhizoma, Ligustici Rhizoma, and Paeoniae Lactiflorae Radix. It has been used clinically for hundreds of years in China to cure females of dysmenorrhea, anemia, and debility etc. Studies showed that water extracts of Siwu are rich in metal elements, such as iron, copper, and zinc (Lu et al., 1990; Lu et al., 1991; Yuan et al., 1991). The velocity of isovolumic contraction of cardiac muscle, the maximum rate of left ventricular pressure change and left ventricular systolic pressure were increased by Siwu tang in normal rats (Li and Zheng, 1995). Ozone-induced free radical damage in mice could be relieved by a Siwu decoction (Gong et al., 1993). The Siwu decoction that was reported to enhance the activity of [Na.sup. ], [K.sup. ]-ATPase and [Ca.sup.2 ], [Mg.sup.2 ]-ATPase in RBC membranes in blood deficiency patients was associated with the effect of enriching the blood (He et al., 1996). These studies indicated that Siwu could nourish the body, promote blood circulation, delay aging, and overcome weakness.

Jiawey Siwu (JS) is a modified formula of Siwu, with the addition of Ginseng Radix and Eucommiae Cortex. Motor activity was enhanced in mice after long-term administration of ginseng (Itoh et al., 1989). The biosynthesis of serum protein was also stimulated in mice with administration of a ginseng saponin mixture (Oura et al., 1975). Both stimulatory and inhibitory effects on the conditioned avoidance response in rats have been shown with Ginseng treatment (Saito et al., 1977). The extract of Eucommiae Cortex was reported to effectively increase collagen synthesis in a false ageing rat model (Li et al., 2000). However, whether the combination of the six herbal drugs in JS is an effective tonic for CNS function and physical capacity remains unclear. In the present study, we investigated the CNS excitatory-inhibitory effect of JS in mice after acute oral administration using a pentobarbital-induced sleep method. We also investigated subchronic (12 day) and chronic (8 week) effects of JS on motor activity and physical endurance in rats by using an activity cage and a rotarod, respectively.

Materials and Methods

Animals

Male Sprague-Dawley rats, 4 weeks old (110-150 g), were used to study ambulatory activity and endurance. Male ICR (Institute for Cancer Research) mice (20-25 g) were used to study sedation. The animals were obtained from the Laboratory Animal Center of National Yang-Ming University in Taiwan. The animals were kept at constant temperature (25 [ or -] 1 [degrees] C), and a solid diet and tap water were supplied ad libitum.

Rats were randomly assigned to 4 groups of control, and 1-, 5-, and 20-times human dosage; they were given same volume normal saline (10 ml/kg) or JS (0.21, 1.05, or 4.2 g/kg) orally once a day for 8 weeks, respectively. Mice were also randomly assigned to four groups (control, 1-, 5-, and 20-times human dosage) for the sedation experiment.

Composition and Preparation of JS

JS pills are the products of Hong Shing Pharmaceutical Plant Co., Ltd. The composition of JS is listed in Table 1. JS was dissolved in distilled water for administration.

Analyses the Maker Components in JS

Ferulic acid, tetramethylpyrazine, paeoniflorin and ginsenoside-[Rg.sub.1] were the maker components of Angelicae Sinensis Radix, Ligustici Rhizoma, Paeoniae Lactiflorae Radix and Ginseng Radix respectively. These compounds were analyzed by high performance liquid chromatography (HPLC) in compared with standard. A high-pressure pump (L-7100, Hitachi, Tokyo, Japan) with interface (D-7000) was connect to a column (Mightysil RP-18, GP 250 x 4.6 mm) and an ultraviolet detector (Hitachi L-7420). The mobile phases were as follows: acetonitril (20%), 0.1% [H.sub.3]P[O.sub.4] (80%) for ferulic acid or paeoniflorin assay; acetonitril (20%), [H.sub.2]O (80%) for tetramethylpyrazine assay; acetonitril (25%), [H.sub.2]O (75%) for ginsenoside-[Rg.sub.1] assay. The flow rate was 1 ml/min. Sample was prepared by powdering the pills and passing through the mesh (60 mesh). Fine powder was dissolved in methanol and shaken at 200 rpm for 60 min. The suspension was then centrifuged in 6000 rpm for 5 min to separate the supernatant. The pellet was repeated to extract as above. The supernatant was combined and passed through a 0.45 [micro]m membrane for HPLC analysis. Ferulic acid, tetramethylpyrazine, paeoniflorin and ginsenoside-[Rg.sub.1] were detected at UV spectra 320 nm, 280 nm, 230 nm and 203 nm, respectively.