We investigated the effects of three traditional Japanese medicines (TJMs), orengedokuto, goreisan and shichimotsukokato on the onset of stroke and survival ratio in stroke-prone spontaneously hypertensive rats (SHRSP), and examined the expression patterns of plasma proteins before and after the onset of stroke. Thirty-two 7-week-old male SHRSP were randomly assigned to four groups. The control group received distilled water. Rats in the orengedokuto, goreisan and shichimotsukokato groups received distilled water containing 0.3% (wt/wt) orengedokuto, goreisan and shichimotsukokato extracts, respectively, from 9 to 20 weeks old. Blood pressure was measured at 12 and at 14 weeks old. Body weight was measured and blood samples were obtained weekly. Plasma samples were analyzed by ProteinChip technology. As for body weight loss after stroke onset, mean body weight in the control group decreased after 14 weeks old, but those in the orengedokuto, goreisan and shichimotsukokato groups showed no decrease at 20 weeks old. Systolic blood pressure showed no significant differences among the four groups at 14 weeks old. The survival ratios of the orengedokuto, goreisan and shichimotsukokato groups were significantly enhanced compared to the control group. Analysis of plasma proteins showed changes in 15 peaks between before and after the onset of stroke in the 3,000-30,000 Da mass ranges in the control group. Especially the peaks at m/z 9,330, 9,480 and 9,700 remarkably decreased after the onset of stroke in the control group. These peaks were identified by western blot analysis as haptoglobin. Interestingly, the decrease was partially prevented by the administration of three TJMs. These results suggest that orengedokuto, goreisan and shichimotsukokato suppress the onset of stroke in SHRSP independently from the mechanism of an anti-hypertensive effect. At the same time, TJMs affect the expression of proteins associated with the onset of stroke in SHRSP.
Accumulating evidence suggests the anti-allergic effect of green tea extract. Although histamine is a major chemical mediator in the pathogenesis of allergic rhinitis, the effect of green tea extract on histamine signaling was unknown. In this study, we investigated the effect of tea extract on the toluene-2, 4-diisocyanate (TDI)-induced up-regulations of the histamine H1 receptor (H1R) and Th2 cytokines gene expressions in the nasal mucosa of TDI-sensitized allergy model rats. Pre-treatment with tea extract once daily for 3 weeks significantly suppressed the TDI-induced mRNA elevations of Th2 cytokines such as interleukin (IL)-4, IL-5, IL-9, and IL-13, and tended to suppress H1R mRNA elevation induced by TDI. Further investigations revealed that the 80% ethanol eluate from a TOYOPEARL HW40EC column, in which epigallocatechin-3-O-gallate (EGCG) was a major constituent, inhibited the IgE-stimulated mRNA elevations of Th2 cytokines in RBL-2H3 cells. EGCG suppressed the elevations of IgE-stimulated IL-4 mRNA and phorbol-12-myristate-13-acetate (PMA)-induced H1R mRNA in a dose-dependent manner. In TDI-sensitized rats, pre-treatment with EGCG once daily for 3 weeks decreased the number of sneezes and suppressed the TDI-induced elevations of H1R and IL-4 mRNAs in the nasal mucosa. These findings suggest that EGCG alleviates nasal symptoms by inhibiting histamine signaling as well as IL-4 signaling by suppressing TDI-induced H1R and IL-4 gene up-regulations in TDI-sensitized rats.
Chrysanthemum flower is a herb included in several Kampo formulae such as chotosan. In this study, we investigated the effects of Chrysanthemum flower produced in Hubei (K1) and Guangdong (K2) provinces in China on the blood pressure and expressions of genes encoding nitric oxide synthase (NOS) and muscarinic receptor subtypes in the brain, using stroke-prone spontaneously hypertensive rats (SHR-SP). Administration of test drugs [K1 and K2 extracts: 1 - 2 g (dried herb weight)/kg/day, p.o.; nicardipine (NIC); 100 mg/kg/day, p.o., once daily for 4 weeks] was started when the average blood pressures of the animals reached ≧ 170 mmHg. K1 and K2 (2 g/kg/day), as well as NIC (100 mg/kg/day), produced a significant decrease in the average blood pressure without affecting the heart rate during the 4-week period of drug administration; however, the blood pressures of the drug-treated groups returned to the vehicle-treated control level within a 2-week washout period. Stroke-associated symptoms and changes in gene expression of some hypertension-related biomarkers in the brain were assessed in SHR-SP after re-starting drug administration. K1 (1-2 g/kg/day), K2 (1-2 g/kg/day), and NIC (100 mg/kg/day) significantly reduced the incidence of stroke-associated symptoms in SHR-SP. Moreover, the expression levels of genes encoding eNOS and nNOS, and M3 and M5 muscarinic receptors, but not those of genes encoding iNOS and M2 and M4 muscarinic receptors in the brain were significantly increased by K1 (1 g /kg/day), K2 (2 g /kg/day), and NIC (100 mg/kg/day). These results suggest that Chrysanthemum K1 and K2 exhibit antihypertensive effects in a SHR-SP model and that the elevation of genes for NOS systems and M3 and M5 muscarinic receptors is relevant to the antihypertensive effects of this herb.
The present study was carried out to determine the effect of hydroalcoholic extract of Tylophora indica (HETI) on experimentally induced myocardial infarction (MI) in rats. Albino rats were treated with HETI at two different doses of 100 mg/kg, (HETI-100), 200 mg/kg (HETI-200) and propranolol 10 mg/kg (PRO-10) for 30 days orally. The hearts were excised and mounted on modified langendorff setup and subjected to 15 min global no flow ischemia and reperfused for 15 min for inducing ischemia-reperfusion injury (IRI). Pretreatment of animals with PRO-10 and HETI-200 provided significant protection to myocardium from IRI damage as indicated by significant decrease in LDH and CK-MB activities in perfusate and an increase in activities of these enzymes in heart tissue homogenate (HTH). Similarly, the recovery (%) in developed tension and heart rate were significantly high in HETI-200 and PRO-10 during post-ischemia when compared to control. Moreover, HETI-200 significantly increases endogenous antioxidants (SOD and catalase) activities when compared to IRI control. The protection offered by HETI could be attributed to the presence of flavanoids which shows antioxidant effect by either inhibiting the release of OFR or enhancing the synthesis of endogenous antioxidants such as SOD and catalase in IRI induced cardiotoxicity. These biochemical findings were further confirmed by histological investigations.
Liquiritigenin is a kind of flavonoid extracted from licorice. To study its anti-tumor effects and mechanisms, SMMC-7721 cells were treated with liquiritigenin at serial concentrations. MTT assay was used to examine the growth inhibitory effects of liquiritigenin. Apoptotic morphological were detected using Hoechst 33258 staining. The apoptosis rate was determined by flow cytometry. P53, bcl-2 and procaspase-3 were analyzed using the Western blot method. The activity of caspase-3 was determined using the caspase-3 activity kit. In the study, liquiritigenin for SMMC-7721 cells growth showed a strong inhibitory effect in a dose-and time-dependent manner. Apoptotic morphological changes in the nuclear chromatin of cells were observed, and flow cytometric analysis demonstrated that apoptosis rates in SMMC-7721 cells with liquiritigenin treatment were 7.48%～41.17%. Western blot results revealed that p53 expression level was increased in a dose-dependent manner after liquiritigenin treatment, and liquiritigenin (0.4 mM) increased the expression of p53 approximately 9-flod compared with control group. In contrast, the expression of bcl-2 protein was decreased after liquiritigenin treatment. The results also revealed that liquiritigenin significantly decreased the expression level of procaspase-3, whereas the activity of caspase-3 increased. The present study indicated that liquiritigenin, probably through the regulation of p53 and bcl-2, inhibited cell proliferation and induced apoptosis for SMMC-7721 cells followed by their down-stream molecular caspase-3 activation.
Cistanche salsa (C.A. Meyer) G. Beck, belonging to the family Orobanchaceae is a parasitic plant that is used as an oriental medicinal tonic in Japan. In a previous study, we reported the immunomodulatory effect of C. salsa extract dialysate (CSD) on the human B-cell lines BALL-1 and Namalwa. In this study, we investigated the effects of CSD on 18 cell lines to obtain more information, with protein-bound polysaccharide-K (PSK) as a reference. We found that with regard to cell growth and antibody production, the responses of almost all cell lines to CSD and PSK were identical. CSD inhibited cell proliferation in several cell lines, including Namalwa. In contrast, it induced cell proliferation in other cell lines, including BALL-1 and T-cell lines. With regard to antibody production, CSD and PSK enhanced IgM production in BALL-1 and IgG production in the plasma B-cell line HMy-2. Furthermore, on gel filtration column chromatography, the same CSD fraction inhibited cell proliferation in Namalwa and the mouse melanoma cell line B16. These results suggest that C. salsa is a biological response modifier similar to PSK and exerts an immunomodulatory effect on both cellular and humoral immunity, and a direct anti-cancer effect.