Saikosaponin D is an agonist of the glucocorticoid receptor (GR), and our preliminary study showed that it possesses neuroprotective effects in corticosterone-treated PC12 cells. However, further proof is required, and the molecular mechanisms of this neuroprotection remain unclear. This study sought to further examine the cytoprotective efficiency and potential mechanisms of action of Saikosaponin D in corticosterone-treated PC12 cells. The cells were treated with 250 μM corticosterone in the absence or presence of Saikosaponin D for 24 h; cell viability was then determined, and Hoechst 33342/propidium iodide (PI) and annexin/PI double staining, and TUNEL staining were performed. Next, mPTP, MMP, [Ca(2+)]i, translocation of the GR to the nucleus and Western blot analyses for caspase-3, caspase-9, cytochrome C, GR, GILZ, SGK-1, NF-Κb (P65), IκB-α, Bad, Akt, Hsp90 and HDAC-6 were investigated. The neuroprotective effects of Saikosaponin D were further confirmed by Hoechst 33342/PI, annexin/PI and TUNEL staining assays. These additional data suggested that Saikosaponin D partially reversed the physiological changes induced by corticosterone by inhibiting the translocation of the GR to the mitochondria, restoring mitochondrial function, down-regulating the expression of pro-apoptotic-related signalling events and up-regulating anti-apoptotic-related signalling events. These findings suggest that SSD exhibited its anti-apoptotic effects via differential regulation of mitochondrial and nuclear GR translocation, partial reversal of mitochondrial dysfunction, inhibition of the mitochondrial apoptotic pathway, and selective activation of the GR-dependent survival pathway.

While the exact cause of systemic lupus erythematosus (SLE) is still unknown, modern medicine has a number of effective treatments for this complex disorder. Corticosteroid hormones help reduce inflammation, antimalarial treatments address flare-ups and immunosuppressive medications work to keep the immune system in check. All these therapies are well tolerated, but accompany an increased risk of infection and nephrotoxicity. Recently, several studies showed that a number of natural and herbal products may also help some SLE patients deal with the debilitating symptoms. In this brief report, we proposed a traditional Chinese medicinal herb--Saikosaponins, and discussed its potential as a treatment option for SLE.

To investigate the suppressive effect of saikosaponin-d (SSd) on hepatic fibrosis in rats induced by CCl(4) injections in combination with alcohol and high fat, low protein feeding and its relationship with the expression of nuclear factor-kappaB (NF-kappaB), tumor necrosis factor-alpha (TNF-alpha) and interleukins-6 (IL-6).

Hepatic fibrosis models were induced by subcutaneous injection of CCl(4) at a dosage of 3 mL/kg in rats. At the same time, rats in treatment groups were injected intraperitoneally with SSd at different doses (1.0, 1.5 and 2.0 mg/kg) once daily for 6 wk in combination with CCl(4), while the control group received olive oil instead of CCl(4). At the end of the experiment, rats were anesthetized and killed (except for 8 rats which died during the experiment; 2 from the model group, 3 in high-dose group, 1 in medium-dose group and 2 in low-dose group). Hematoxylin and eosin (HE) staining and Van Gieson staining were used to examine the changes in liver pathology. The levels of alanine aminotransferase (ALT), triglyeride (TG), albumin (ALB), globulin (GLB), hyaluronic acid (HA) and laminin (LN) in serum and the content of hydroxyproline (HYP) in liver were measured by biochemical examinations and radioimmuneoassay, respectively. In addition, the expression of TNF-alpha and IL-6 in liver homogenate was evaluated by enzyme-linked immunosorbent assay (ELISA) and the levels of NF-kappaBp65 and I-kappaBalpha in liver tissue were analyzed by Western blotting.

Both histological examination and Van Gieson staining demonstrated that SSd could attenuate the area and extent of necrosis and reduce the scores of liver fibrosis. Similarly, the levels of ALT, TG, GLB, HA, and LN in serum, and the contents of HYP, TNF-alpha and IL-6 in liver were all significantly increased in model group in comparison with those in control group. Whereas, the treatment with SSd markedly reduced all the above parameters compared with the model group, especially in the medium group (ALT: 412 +/- 94.5 IU/L vs 113.76 +/- 14.91 IU/L, TG: 0.95 +/- 0.16 mmol/L vs 0.51 +/- 0.06 mmol/L, GLB: 35.62 +/- 3.28 g/L vs 24.82 +/- 2.73 g/L, HA: 42.15 +/- 8.25 ng/mL vs 19.83 +/- 3.12 ng/mL, LN: 27.56 +/- 4.21 ng/mL vs 13.78 +/- 2.57 ng/mL, HYP: 27.32 +/- 4.32 mug/mg vs 16.20 +/- 3.12 mug/mg, TNF-alpha: 4.38 +/- 0.76 ng/L vs 1.94 +/- 0.27 ng/L, IL-6: 28.24 +/- 6.37 pg/g vs 12.72 +/- 5.26 pg/g, respectively, P < 0.01). SSd also decreased ALB in serum (28.49 +/- 4.93 g/L vs 37.51 +/- 3.17 g/L, P < 0.05). Moreover, the expression of NF-kappaB p65 in the liver of treated groups was lower than that in model groups while the expression of I-kappaBalpha was higher in treated group than in model group (P < 0.01). The expression of NF-kappaBp65 and TNF-alpha had a positive correlation with the level of HA in serum of rats after treatment with CCl(4) (r = 0.862, P < 0.01; r = 0.928, P < 0.01, respectively).

SSd attenuates CCl(4)-induced hepatic fibrosis in rats, which may be related to its effects of hepato-protective and anti-inflammation properties, the down-regulation of liver TNF-alpha, IL-6 and NF-kappaBp65 expression and the increased I-kappaBalpha activity in liver.

Disruption of the hypothalamo-pituitary-adrenal (HPA) axis characterized by dysfunction of the glucocorticoid negative feedback system is frequently observed in human depressives and is thought to involve a reduction in glucocorticoid receptor (GR) function in the feedback sites including the brain. Recently, we found that chronic stress in rats induces similar HPA disruption that is caused by abolishment of feedback ability in the prefrontal cortex (PFC) and hippocampus, which involves decreased cytosolic GRs or increased nuclear GRs, respectively. Also, we found that saikokaryukotsuboreito (SRBT), a herbal medicine, prevents the chronic stress-induced HPA disruption. We therefore examined here the effects of this drug on the chronic stress-induced changes in GRs in the PFC and hippocampus. Chronic stress was induced in rats by water immersion and restraint (2 h/day) for 4 weeks. SRBT significantly prevented decreased cytosolic GRs in the PFC and increased nuclear GRs in the hippocampus in the chronically stressed rats. Moreover, SRBT significantly prevented the abolishment of feedback ability in both regions. These results suggest that the beneficial effects of SRBT on the GR level are involved in its ameliorating actions on the HPA disruption. This finding provides information important for the prevention and treatment of depression.

The effects of saikosaponin-d, a triterpene saponin derived from Bupleurum falcatum L. (Umbelliferae), on the signaling pathways of T cell activation were examined. The results showed that saikosaponin-d potently suppressed both early (CD69) and late (CD71) expressions of mouse T cells stimulated with Con A or PMA. It interfered with PKCtheta translocation from cytosol to membrane fraction and inhibited the phosphorylations of IkappaBalpha and JNK, but not ERK, in PMA-activated mouse T cells. Additionally, it inhibited PMA and ionomycin-stimulated IL-2 production in mouse T cells. In summary, these results indicate that the mechanism by which saikosaponin-d inhibits T cell activation would involve the suppression of CD69 and CD71 expressions and IL-2 production, and the modulation of PKC pathway through PKCtheta, JNK, and NF-kappaB transcription factor. This may herald a novel approach for further studies of saikosaponin-d as a candidate for use in the treatment of inflammatory and autoimmune diseases.

Mesangioproliferative glomerulonephritis is a common kidney disease and at present, there is no effective treatment. Our previous studies have demonstrated that Sairei-to can significantly prevent progression of experimental glomerulonephritis in rats. Although we have reported that the active component of Sairei-to in treatment of glomerulonephritis was Saikosaponin-d (Ssd), mechanism of Ssd in prevention of mesangioproliferative glomerulonephritis progression is still unknown. Therefore, current study examines the effects of Ssd on progression of mesangioproliferative glomerulonephritis induced by anti-Thy1 monoclonal antibody 1-22-3 (mAb 1-22-3) in uninephrectomized rats.

Eighteen female Wistar rats first received uninephrectomy and mAb 1-22-3 injection and were then divided into 3 groups: treated daily with phosphate-buffered saline (PBS), 0.6 or 1.8 mg/kg of Ssd. Urinary protein concentration and systolic blood pressure were evaluated and the kidneys were collected and subjected to histological and immunohistological evaluation. The mRNA and protein of the kidneys were extracted and subjected to reverse transcriptase polymerase chain reaction and Western blot analysis, respectively.

Ssd reduced the amount of urinary protein and systolic blood pressure. Ssd administration also decreased extracellular matrix expansion, crescentic formation as well as infiltration of macrophages and CD8+ T lymphocytes. Moreover, Ssd significantly reduced expression of transforming growth factor beta 1 (TGF-beta1) and type I collagen in the kidneys.

Ssd inhibits the progression of mesangioproliferative glomerulonephritis through reduction of the expression of TGF-beta1 and the infiltration of macrophages and CD8+ T lymphocytes.

Sho-hange-ka-bukuryo-to and Nichin-to, traditional Chinese herbal (Kampo) medicines have been used to treat vomiting and nausea. Traditional herbal medicines have frequently been used in the empirical treatment. Some patients who take these medicines have no organic disease but have conditions classified as non-ulcer dyspepsia (NUD). To determine the pharmacological effects of Sho-hange-ka-bukuryo-to, Nichin-to, and the two herbs (Pinelliae Tuber and Zingiberis Rhizoma, both of which are included in Sho-hange-ka-bukuryo-to and Nichin-to), we examined the effects of these medicines on the plasma levels of adrencorticotropic hormone (ACTH) and cortisol under stress conditions by repetitive blood sampling. After a single administration of Kampo medicine or a placebo, venous blood samples were taken before and 20-240 min after administration. A single administration of Sho-hange-ka-bukuryo-to caused significant suppression of an increase in plasma ACTH-immunoreactive substance (IS) levels at 120 to 180 min and tended to suppress increases in plasma cortisol levels at 240 min, compared with the response to a placebo. A single administration of Nichin-to caused significant suppression of increases in plasma ACTH-IS levels at 120 min compared with a placebo group, but had no effect on plasma cortisol levels. Pinelliae Tuber had no significant effects in plasma ACTH-IS or cortisol, but Zingiberis Rhizoma significantly suppressed the increase of ACTH-IS (120 min) and cortisol (180 min). These medicines have a modulatory effect on the hypothalamo-pituitary-adrenal (HPA) axis and autonomic nervous function. These effects might be beneficial in stress-related disease and suggest that this medicine has clinical pharmacological activity.

Platycodon root, one of the most important Chinese herbal medicines, has been used as an antiphlogistic, antitusivie, and expectorant agent since ancient times. In the Japanese Pharmacopoeia XIV this is listed as the root of Platycodon grandiflorum A. De Candolle (Campanulaceae) and called KIKYOU (Platycodi Radix) in Japanese. HPLC analysis showed that commercial samples of P. Radix all contained platycodins, and a total of 12 peaks were identified by co-HPLC analysis with authentic samples isolated earlier from this laboratory. The peak purity and identity were checked with a photodiode array detector. The contents of the major saponins, platycodins A, C, and D, were determined and the peak-area ratios of platycodins A, C, and D, were shown to be correlated with their sources of origin. Fourteen commercial samples of Platycodon root, the origin of which was Platycodon grandiflorum, were collected from China (5 samples), Korea (5 samples), and Japan (4 samples). The commercial samples from China, Korea, and Japan each gave a distinct HPLC pattern with peak-area ratio of platycodins A, C, and D, of 1:2:3 and 2:8:1, respectively. HPLC analysis showed that those on the Japanese market were either imported from China or Korea based upon their HPLC patterns.

Exposure to chronic stress is thought to play an important role in the etiology of depression. This disorder has been shown to involve disruption of the hypothalamo-pituitary-adrenal (HPA) system and dysfunction of the prefrontal cortex (PFC). We have demonstrated that chronic stress in rats induces similar HPA disruption or a depressive state caused by a reduction of dopaminergic and serotonergic transmission in the PFC. We have also shown that saiko-ka-ryukotsu-borei-to, a herbal medicine, prevents such chronic stress-induced HPA disruption. However, the behavioral and neurochemical bases of this drug remain unclear. Here we examined the effects of saiko-ka-ryukotsu-borei-to on the depressive behavioral state and the reduction of transmission resulting from chronic stress. The chronic stress was induced by water immersion and restraint (2 h/day) for 4 weeks followed by recovery for 10 days. The treatment with saiko-ka-ryukotsu-borei-to (100, 300, or 1000 mg/kg p.o.) ameliorated the stress-induced depressive state in a dose-dependent manner, evaluated by a rotarod test. A microdialysis study indicated that the drug treatment significantly prevented the chronic stress-induced decreases in extracellular concentrations of dopamine and serotonin in the PFC. These results suggest that saiko-ka-ryukotsu-borei-to ameliorates the chronic stress-induced depressive state based on the prevention of PFC dysfunction. These findings provide important information for treatment of depression.

Rikkunshi-to, a gastrointestinal function regulatory traditional Chinese herbal (Kampo) medicine, has recently been evaluated for its clinical usefulness in stress and depression. This medicine has modulatory effects on the hypothalamo-pituitary-adrenal axis and autonomic nervous function. We examined the effect of Rikkunshi-to and the other gastrointestinal function regulatory Kampo medicines, Hange-shashin-to, Hange-koboku-to, and Ninjin-to, on the plasma levels of adrenocorticotropic hormone (ACTH) and cortisol under stress conditions by repetitive blood sampling. Rikkunshi-to, Hange-shashin-to, and Hange-koboku-to significantly suppressed increases in plasma ACTH-immunoreactive substance (IS) levels compared with the response to a placebo. Rikkunshi-to and Hange-shashin-to significantly suppressed increases in plasma cortisol levels compared with the response to placebo. Ninjin-to had no significant effect on plasma ACTH-IS and cortisol levels. In this study, Rikkunshi-to, Hange-shashin-to, and Hange-koboku-to (partially) regulated plasma ACTH and cortisol levels under stress. These modulatory effects might be beneficial in stress-related disease and suggest that these medicines have clinical pharmacologic activity.

Exposure to stress is known to precipitate or exacerbate many neuropsychiatric disorders such as depression. Abnormality of the neuroendocrine system, as shown by increased adrenal weight and attenuated glucocorticoid negative feedback, is frequently seen in depression. The aim of the present study is to clarify the usefulness of saiko-ka-ryukotsu-borei-to, an herbal medicine, in the treatment of abnormality of the neuroendocrine system using an experimental stress-depression model. Rats were subjected to water immersion and restraint for 2 h daily for 4 weeks (chronic stress), followed by recovery for 10 days. Saiko-ka-ryukotsu-borei-to was administered during the stress and recovery periods (100, 300, or 1000 mg/kg daily, p.o.) or only during the recovery period (1000 mg/kg). After the recovery period, the adrenal weight was measured, and glucocorticoid feedback ability was evaluated by a dexamethasone suppression test using 30 microg/kg dexamethasone. The administration of saiko-ka-ryukotsu-borei-to during the stress and recovery periods prevented the stress-induced increase in adrenal weight or the attenuated negative feedback in a dose-dependent manner. The administration of saiko-ka-ryukotsu-borei-to during the recovery period alone also ameliorated the abnormality of the neuroendocrine system. These results indicate that saiko-ka-ryukotsu-borei-to is effective against chronic stress-induced abnormality of the neuroendocrine system. Because some symptoms and symptomatic relapses in depressives are attributed to dysfunction of the hypothalamo-pituitary-adrenal axis, the present findings provide information important for prevention and treatment of depression.

This paper reviews the role of herbal-based medicines in the treatment of asthma and allergic rhinitis. A comprehensive literature search was performed of relevant English-language papers and abstracts were identified through a MEDLINE search and from bibliographies of the identified papers. Papers and studies pertaining to the use of medicinal plants in the treatment of asthma and allergic rhinitis were identified. They were then analyzed according to design, inclusion and exclusion criteria, population studied, variables tested, method of treatment (i.e., specific medicinal plants or herbal combinations), and results. The data have been reviewed and divided on the basis of culture and the effects of medicinal plants in asthma and allergy. A number of studies were found that support the use of some herbal medicines in asthma and allergy. Various derivatives from specific medicinal plants were identified as the antiasthma components and some mechanisms of action were explored. The results show positive effects of these herbs on bronchodilation, pulmonary function tests, and antagonism of asthma mediators such as histamine and platelet activating factor, corticosteroid levels, and clearance of mucus. Improved symptoms were also seen in patients with allergic rhinitis specifically on histamine-induced reactions, e.g., rhinorrhea, sneezing, and itching. From the review, there has been a role for some herbal medicines in the treatment of asthma and allergic rhinitis. Usage of herbal medicines has increased in recent years. Many of these medicinal plants provided relief of symptoms equal to allopathic medicines used. Specific chemical derivatives have been isolated from many of these plant products which act on the mechanisms and mediators that cause asthma and allergies. The amount of research on these products, especially in the United States, is limited. There is a lack of control of quantity and quality of the components in these remedies. Yet, many have fewer side effects than current therapy. Throughout the history of medicine, drugs have been developed from traditional medicine. By continuing to investigate how some of these herbal interventions work, we may be able to find additional effective medicines to treat asthma and allergies.

High titer rabbit polyclonal antibodies (pAbs) which show a specificity for saikosaponin a (SSA), have been generated. The immunogen used was a conjugate of SSA linked through its glucose moiety to bovine serum albumin by periodate oxidation method. The antibody titers obtained from two rabbits, inoculated with the immunogen, reached a plateau after the fourth and third booster injection, respectively. The specificity of the pAbs was determined by hapten inhibition assays using several SSA-like structures. SSA competitively inhibited the binding of the rabbit anti-SSA pAbs to SSA-ovalbumin on solid phase, a coated antigen on the well. The antibodies showed high specificity to SSA, exhibiting no significant cross-reactivity with any of SSA analogues tested.

The effects of traditional Chinese medicine (Sairei-to) on experimental glomerulonephritis induced in rats by monoclonal antibody (mAb) 1-22-3 injection was examined. The level of proteinuria in the Sairei-to-treated group was significantly lower than that in the PBS treated group. This suppressive effect was caused by the major component of Sairei-to, Syo-saiko-to but not by another component, Gorel-san. The suppressive effect of Syo-saiko-to was identified in its components (Bupleuri radix, Pinelliae tuber and Zingiberis rhizoma), but not in the other combined components (Ginseng radix and Zizyphi fructus). Further study revealed that the suppressive effects of the combined components were mainly derived from Bupleuri radix. It was demonstrated that the actual active ingredient is probably Saikosaponin-d. Light microscopy revealed that Sairei-to and its effective components suppressed the proliferation of mesangial cells and mesangial matrix expansion. Semiquantitative morphological studies of glomerular lesions on the eighth day showed that Syo-saiko-to and its combined components (Bupleuri radix, Zingiberis rhizoma and Pinelliae tuber) suppressed mesangial matrix expansion significantly compared with phosphate-buffered saline control groups (matrix score: 28.0 +/- 19.1 vs 102.3 +/- 14.1; 30.9 +/- 30.1 vs 102.3 +/- 14.1, P < 0.005, respectively). It was concluded that Saikosaponin-d, as well as Bupleuri radix, Syo-saiko-to and Sairei-to can suppress proteinuria and morphological changes in the rat glomerulonephritis model induced by mAb 1-22-3.

Saiko agents, Chinese herbal drugs, stimulate corticotropin-releasing factor (CRF) release from the hypothalamus, adrenocorticotropin (ACTH) secretion and proopiomelanocortin (the precursor for ACTH) gene expression in the anterior pituitary. In the present study, the effect of intracerebroventricular injection of saikosaponin (SS)-a and -d, two of the main components of saiko agents, on hypothalamic CRF gene expression was examined in pentobarbital-anesthetized rats. Administration of SS-d, 0.2-2.0 micrograms/kg body wt, increased plasma ACTH levels, proopiomelanocortin mRNA levels in the anterior pituitary and the CRF mRNA level in the hypothalamus in a dose-dependent manner, whereas SS-a failed to have an affect on these levels. These findings indicate that SS-d stimulates both CRF gene expression and CRF release, which in turn increases ACTH release and proopiomelanocortin gene expression in the anterior pituitary. Therefore, SS-d is believed to have an important role both in saiko agent-induced CRF release and CRF gene expression in the hypothalamus.

Three major traditional Chinese medicines (TCM), Sho-saiko-To, Saiboku-To, and Sairei-To, consist of similar herbal prescriptions containing glycyrrhizin, which is a strong inhibitor of 11 beta-hydroxysteroid dehydrogenase. We performed cross-over open trials in healthy subjects to clarify prednisolone pharmacokinetics on co-administration of these preparations. All subjects received a single oral dose of 10 mg prednisolone before oral treatment with one of the test preparations. After a 2-week wash-out interval, they received one of the test preparations for three days at daily doses of 7.5 or 9.0 g. On the third study day, 10mg prednisolone was administered orally in combination with the test preparation. Area under the curves (AUC) of prednisolone before and after the treatment decreased from 0.94 to 0.78 mg h L-1 (P < 0.05) in the Sho-saiko-To group, increased from 0.92 to 1.06 mg h L-1 (P < 0.01) in the Saiboku-To group, and did not change in the Sairei-To group. AUC ratios of prednisone and prednisolone, which reflect the 11 beta-hydroxysteroid dehydrogenase activity, increased in the Sho-saiko-To group (P < 0.01), decreased in the Saiboku-To group (P < 0.01), and did not change in the Sairei-To group after the treatments. Similar results were observed in ratios of endogenous cortisone to cortisol. Because of the equal glycyrrhizin content in all three preparations, it was unexpected that the 11 beta-hydroxysteroid dehydrogenase effect was different amongst the three groups. These observations suggest that some unknown metabolic enzyme modifiers, promoters or inhibitors, may be involved in these traditional treatments.

We examined the immunoregulatory action of saikosaponin-d (SSd), which was isolated from the root of Bupleurum talcatum L. and had a steroid-like structure, on murine thymocytes, and compared the action with that on spleen cells. Constitutive DNA synthesis or the growth response stimulated with anti-CD3mAb of thymocytes were down-regulated by 3 micrograms/ml SSd, whereas with spleen cells these were up-regulated by the same concentration of SSd. On the other hand, 3 micrograms/ml of SSd greatly up-regulated the growth response and interleukin 2 (IL-2)/interleukin 4 (IL-4) production induced through a receptor-bypassed pathway by calcium ionophore A23187 plus phorbol 12-myristate 13-acetate (PMA) in thymocytes, whereas it only slightly up-regulated them in spleen cells. Moreover, the same concentration of SSd inhibited DNA fragmentation in thymocytes induced by A23187 or PMA. These results suggest a unique cell type-dependent immuno-modulatory action of SSd.

To identify the inhibitor of prednisolone metabolism contained in Saiboku-To, we conducted in-vitro experiments of 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD), using rat liver homogenate and cortisol as a typical substrate. We studied the effects of ten herbal constituents on 11 beta-HSD. Five herbal extracts showed inhibitory activity with Glycyrrhiza glabra > Perillae frutescens > Zizyphus vulgaris > Magnolia officinalis > Scutellaria baicalensis. This suggests that unknown 11 beta-HSD inhibitors are contained in four herbs other than G. glabra which contains a known inhibitor, glycyrrhizin (and glycyrrhetinic acid). Seven chemical constituents which have been identified as the major urinary products of Saiboku-To in healthy and asthmatic subjects were studied; magnolol derived from M. officinalis showed the most potent inhibition of the enzyme (IC50, 1.8 x 10(-4) M). Although this activity was less than that of glycyrrhizin, the inhibition mechanism (non-competitive) was different from a known competitive mechanism. These results suggest that magnolol might contribute to the inhibitory effects of Saiboku-To on prednisolone metabolism through inhibition of 11 beta-HSD.

Administration of Saireito, a Saiko agent (a Chinese herbal drug), via a stomach cannula stimulates ACTH release and proopiomelanocortin, the precursor for ACTH, gene expression in the rat anterior pituitary. To study whether Saireito-stimulated secretion and synthesis of ACTH are mediated by hypothalamic corticotropin-releasing factor (CRF), we examined the effect of passive immunization of endogenous CRF by i.v. administration of CRF antiserum on Saireito-increased plasma ACTH levels and proopiomelanocortin gene expression in the rat anterior pituitary, under pentobarbital anesthesia. CRF antiserum inhibited Saireito-induced plasma ACTH levels and proopiomelanocortin mRNA levels in the anterior pituitary. This result indicates that Saireito stimulates CRF neurons to increase CRF release, which stimulates secretion and synthesis of ACTH.

Saiboku-To, a mixture of ten different herbal extracts, has been used in Japan and Czechoslovakia for corticosteroid-dependent severe asthma to reduce the maintenance doses of corticosteroid. Magnolol has been considered to be an active component of Saiboku-To as an inhibitor of 11 beta-hydroxysteroid dehydrogenase and T-lymphocyte proliferation resulting in corticosteroid-sparing. To investigate the relationship between magnolol and the clinical effects of Saiboku-To, urinary magnolol excretion was compared in responders and non-responders under long-term Saiboku-To treatment. The clinical outcome of the Saiboku-To treatment was evaluated in nine asthmatic patients at 52 weeks after the onset of the treatment, using individual fluctuation of asthmatic points obtained from the patients' diary cards. Three patients whose clinical conditions were improved by the treatment were termed responders and six others were termed non-responders. The difference in the amounts of the total magnolol excreted were not significant; however, free (or non-conjugated) amounts of magnolol excreted in the responders were 7 times those in the non-responders (P < 0.05). These results suggest that the magnolol might be responsible for the therapeutic effect of Saiboku-To, indicating practical bioavailability in the responders.

The effect of administration of Saireito, a Saiko agent, via a stomach cannula on adrenocorticotropin (ACTH) release and gene expression of proopiomelanocortin (POMC), the precursor for ACTH, in the anterior pituitary, as well as on the corticotropin-releasing factor (CRF) in the hypothalamus, was examined in pentobarbital anesthetized rats. Saireito decreased the hypothalamic CRF level due to an early release of CRF and stimulated ACTH release and POMC gene expression but did not increase CRF gene expression. These results suggest that Saireito does not stimulate CRF gene expression, although it does stimulate CRF release, which in turn stimulates POMC gene expression in the anterior pituitary and ACTH release.

A novel strategy for discovering biologically active components in traditional Chinese herb remedies was performed from a pharmacokinetic view. The hypothesis was that the active compounds should appear in blood and urine with appropriate blood concentrations and urinary excretion rates after the administration of herbal-extract mixtures. In this research, we applied our procedures to Saiboku-To, one of the most popular Chinese herbal medicines in Japan. Consisting of 10 different plant extracts, it is used for the treatment of bronchial asthma. The analytical method adopted was a rapid-flow fractionation (RFF) for extraction-fractionation of lipophilic components in urine followed by silica-gel high-performance liquid chromatography (HPLC) equipped with a multichannel ultraviolet (uv) absorption detector. beta-D-Glucuronidase-treated urine samples collected before and after the administration of Saiboku-To to healthy and asthmatic subjects were treated with the RFF apparatus to afford three pH-dependent fractions: strongly acidic (S), weakly acidic (W), and neutral (N). HPLC of these fractions, monitored by the multichannel uv detector, showed three new peaks in the postadministrative urine: one in the N fraction, two in the W fraction, and none in the S fraction. A compound in the N fraction was identified with authentic magnolol, a major component in Magnolia officinalis. Two compounds in the W fraction were identified by comparison with authentic samples as 8,9-dihydroxydihydromagnolol and liquiritigenin, metabolites previously isolated from M. officinalis and Glycyrrhiza glabra, respectively.