Anti-allergic Activity of the Sophorae Radix Water Extract in Experimental Animals

American Journal of Chinese Medicine,  Wntr, 2001  by Lee Ho-sub,  Park Kwan-Ha,  Kwon Kang-Beom,  Mun Byeong-Sun,  Song Cheol-Min,  Song Yong-Seon,  Seo Eun-A,  Kim Young-suk,  Kim Kang-Jeon,  Ryu Do-Gon

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(Accepted for publication August 8, 2000)

Abstract: The water extract of Sophorae radix was tested for its preventive effects against cardiovascular anaphylaxis elicited in experimental animals. In actively sensitized pithed rats, the extract partially protected the animals from death resulting from antigen challenge. Of the cardiovascular changes observed during the anaphylaxis in the pithed animals, the initial pressor response was significantly attenuated by the extract. The extract also improved anaphylactic cardiac dysfunction in passively sensitized isolated guinea hearts: improvement was noted in the contractility, arrhythmic duration and lactate dehydrogenase elevation. The perfusion pressure change to antigen challenge was not altered by the extract in passively sensitized isolated mesenteric artery preparations. The extract however significantly inhibited homologous passive cutaneous anaphylaxis responses in rats. When examined in isolated guinea pig tracheal muscle, the extract did not influence at reasonable concentrations on the contraction elicited by three major anaphylactic mediators, histamine, leukotriene D4 and serotonin. These results suggest that the water extract of Sophorae radix possesses anti-anaphylactic effect in cardiovascular system, primarily acting on the heart not the peripheral resistance vessels.

Sophorae radix, the dried root of Sophora flavescens Aiton, contains numerous pharmacologically active principles. The most intensively studied ones are several antiarrhythmic alkaloids including d-matrine (Xin and Lin, 1987; Zhang et al., 1990). Sophora flavescens extract has been used as an anti-asthmatic, expectorant, diuretic, and natriuretic agent in oriental medicine (Huang, 1993). In relation to immune-associated action of this medicine, oxymatrine, an extracted principle from Sophora flavescens, has been demonstrated to suppress the mouse B cell proliferation without prominent effect on T cells (Qin et al., 1990). In the Hawaiian tradition the extract of Sophora chrysophylla, a close species to Sophora flavescens Aiton, has been used for the treatment of asthma (Massey et al., 1994). Thus although there are implications that Sophora flavescens may be effective against allergic symptoms, no one has ever examined systemically whether the plant is useful for some types of allergy. In particular, in this research, we examined the effect of the plant extract on the cardiovascular anaphylaxis for which any reasonably efficacious medicine has not been reported.

Materials and Methods

Animals

All animals were purchased as specific pathogen-free status (Dai-Han Experimental Animal Center, Seoul, Korea) and maintained under conventional conditions at 21 [+ or -] 1 [degrees] C room temperature and 55 [+ or -] 2% relative humidity. Water and feed were supplied ad libitum.

Preparation of the Extract

For water extraction of Sophora flavescens, 200 g of dried root was mixed with 1,800 ml of distilled water and extracted under reflux for 3 hr by boiling the mixture. Following centrifugation at 3,000 x g for 20 min the supernatant was filtrated with a Whatman filter paper. The filtrate was concentrated to about 300 ml with a rotary evaporator at 60 [degrees] C under vacuum and freeze-dried to dryness.

Pithed Rats

Sprague-Dawley male rats (230-250 g) were sensitized with 10 mg/rat of turkey ovalbumin (OVA; Grade VI, Sigma, St. Louis) administered intraperitoneally for two consecutive days (Sun et al., 1992). On day 21 after the first sensitization, animals were anesthetized with ether and tracheal cannula was inserted into the trachea for artificial respiration (60 strokes/min, 1 ml/100 g body weight). Animals were pithed through the right orbit to the spinal column with a round copper rod of 1.5 mm in diameter (Shipley and Tilden, 1947). The right common carotid artery and jugular vein were respectively cannulated for cardiovascular parameter monitoring and OVA challenge. Animals were maintained at 37 [degrees] C throughout the experiment with a water bath. The test substance was administered orally for three days, on Day 19, 20 and 21. The last dosing was made 1 hr prior to the commencement of surgery. Changes of arterial blood pressure and heart rate were monitored using a pressure transducer and a cardiotachometer, respectively, and recorded with a physiography (Polygraph 4006, Barcelona, Spain).

Guinea Pig Langendorff Hearts

Hartley male guinea pigs weighing 300-350 g were passively sensitized with guinea pig anti-OVA serum by injecting into the peritoneal cavity 24 hr before the experiment. Guinea pigs were sacrificed bleeding through cervical dissection under ether anesthesia and hearts were rapidly removed. An aortic cannula was inserted and attached to a Langendorff apparatus (Park et al., 1990). Hearts were perfused with Krebs-Henseleit solution (in mM, NaCl 118; KCl 4.7; Ca[Cl.sub.2] 2.5; Mg[SO.sub.4] 1.6; Na[HCO.sub.3] 24.9; [KH.sub.2][PO.sub.4] 1.2; glucose 11.0, pH 7.4) under the constant perfusion pressure of 60 cm [H.sub.2]O. The perfusion solution was maintained at 37 [degrees] C and continuously saturated with 95% [O.sub.2]-5% [CO.sub.2]. A nylon thread was connected from the apex of the heart to an isometric transducer to monitor contractility and heart rate. Coronary effluent was collected at 1 min intervals for coronary flow change measurement and an aliquot was used for lactate dehydrogenase (LDH) and creatine kinase (CK) activity determination (Sigma kit). For anaphylaxis induction, 2 mg of OVA was injected into the heart. The test substance was infused for 20 min before the anaphylactic challenge. Edema index was calculated by comparing the wet heart weight right after the experiment to dry weight.