Immunomodulatory effect of Schizonepeta tenuifolia water extract on mouse Th1/Th2 cytokine production in-vivo and in-vitro.

Schizonepeta tenuifolia (ST) is a major herbal constituent included in treatments for the common cold with fever, ostitis media and other skin inflammations. The present study investigated the effect of ST water extract on the pattern of cytokine production from activated T cells in-vivo and in-vitro. When ST (200 mgkg(-1)) was given orally to mice for 7 days before i.v. injection of anti-CD3 antibody, it significantly decreased mRNA levels of interleukin (IL)-4, interferon (IFN)-gamma and T-bet. Our flow cytometric analysis showed that ST administration significantly increased CD69 expression but showed little effect on the subsets of T cells. When we cultured mouse CD4 T cells under Th1/Th2 differentiation in the presence of ST, the suppressive activity of ST on IFN-gamma involved T-bet, but the downregulation of IL-4 occurred independently of the Th2 transcription factors GATA binding protein 3 (GATA-3) and c-Maf. However, it increased IL-2 secretion during Th1/Th2 differentiation. Our study demonstrates that ST regulates inflammatory responses by reducing the release of Th1 and Th2 cytokines from T cells and prevents unprimed CD4 T cells from differentiating into Th1 and Th2 cells.

Molecular changes in remote tissues induced by electro-acupuncture stimulation at acupoint ST36.

To investigate the effects of electro-acupuncture (EA) treatment on regions remote from the application, we measured cellular, enzymatic, and transcriptional activities in various internal tissues of healthy rats. The EA was applied to the well-identified acupoint ST36 of the leg. After application, we measured the activity of natural killer cells in the spleen, gene expression in the hypothalamus, and the activities of antioxidative enzymes in the hypothalamus, liver and red blood cells. The EA treatment increased natural killer cell activity in the spleen by approximately 44%. It also induced genes related to pain, including 5-Hydroxytryptamine (serotonin) receptor 3a (Htr3a) and Endothelin receptor type B (Ednrb) in the hypothalamus, and increased the activity of superoxide dismutase in the hypothalamus, liver, and red blood cells. These findings indicate that EA mediates its effects through changes in cellular activity, gene expression, and enzymatic activity in multiple remote tissues. The sum of these alterations may explain the beneficial effects of EA.

Inhibition of cell cycle progression via p27Kip1 upregulation and apoptosis induction by an ethanol extract of Rhus verniciflua Stokes in AGS gastric cancer cells.

Botanical preparations are widely used by patient with cancer in Korea, Japan and China. Rhus verniciflua Stokes (RVS) has traditionally been used as a medicinal ingredient for the therapy of stomach and uterine cancer. In this study, we showed that exposure to an ethanol extract of RVS (50 microg/ml) resulted in a synergistic inhibitory effect on cell growth in AGS cells. Growth inhibition was related with the inhibition of proliferation and induction of apoptosis. The extract induces G1-cell cycle arrest through the regulation of cyclins, the induction of p27Kip1, and decrease the CDK2 kinase activity. The upregulated p27Kip1 level is caused by protein stability increment by the reduction of Skp2, a key molecule related with p27Kip1 ubiquitination and degradation, and de novo protein synthesis. RVS extract induces apoptosis through the expression of Bax, poly(ADP-ribose) polymerase (PARP) and activation of caspase-3. RVS extract induces G1-cell cycle arrest via accumulation of p27Kip1 controlled by Skp2 reduction and apoptosis passing through an intrinsic pathway in human gastric cancer cells but not in normal cells, therefore we suggest that this extract could be a candidate medicine or compound for the development of novel class of anti-cancer drugs.

Saussurea lappa induces G2-growth arrest and apoptosis in AGS gastric cancer cells.

The molecular effects of Saussurea lappa extracts, a traditional medicine in Eastern Asia, on the fate of gastric carcinoma have not been understood. In this study, its cytostatic effects were examined using gastric AGS cancer cells. Its treatment resulted in apoptosis and G2-arrest in a dose- and time-dependent manner. The effects were attributed to the regulation of cyclins and pro-apoptotic molecules and suppression of anti-apoptotic molecules. Therefore, these results suggest that extracts of S. lappa root may be a candidate to deal with gastric cancers either by traditional herbal therapy or by combinational therapy with conventional chemotherapy.

Immunomodulatory effect of Astragali Radix extract on murine TH1/TH2 cell lineage development.

Astragali Radix (AR), is a popular herbal medicine used to treat allergic diseases in Korea, Japan and China. Our study examined the effect of an AR ethanol extract on both in vitro and in vivo murine CD4 T cells' differentiation into Th1 and Th2 subsets. CD4 T cells from Balb/c mice were activated with anti-CD3/anti-CD28 mAb in the presence of AR for 2 d. AR treated cells showed an elevated level of IL-4 but a reduced level of IFN-gamma secretion. In addition, in vitro Th1/Th2 polarization experiments revealed that AR enhanced the levels of IL-4 in Th2 cells but reduced the levels of IFN-gamma in Th1 cells. To elucidate the effects of AR in Th1/Th2 lineage development during the in vivo condition, AR was administrated orally to BALB/c mice. The results demonstrated that AR administration significantly increased IL-4 production in both the serum and supernatant of splenocyte culture, while IFN-gamma secretion was diminished upon in vivo activation with anti-CD3 antibody. Our data clearly indicates that AR selectively alters Th1/Th2 cytokine secretion patterns and provides the pharmacological basis for AR's clinical applications.

Induction of apoptosis by Saussurea lappa and Pharbitis nil on AGS gastric cancer cells.

We performed this study to understand the molecular basis underlying the antitumor effects of Saussurea lappa, Pharbitis nil, Plantago asiatica and Taraxacum mongolicum, which have been used for herbal medicinal treatments against cancers in East Asia. We analyzed the effects of these medicinal herbs on proliferation and on expression of cell growth/apoptosis related molecules, with using an AGS gastric cancer cell line. The treatments of Saussurea lappa and Pharbitis nil dramatically reduced cell viabilities in a dose and time-dependent manner, but Plantago asiatica and Taraxacum mongolicum didn't. FACS analysis and Annexin V staining assay also showed that both Saussurea lappa and Pharbitis nil induce apoptotic cell death of AGS. Expression analyses via RT-PCR and Western blots revealed that Saussurea lappa, but not Pharbitis nil, increased expression of the p53 and its downstream effector p21Waf1, and that the both increased expression of apoptosis related Bax and cleavage of active caspase-3 protein. We also confirmed the translocation of Bax to mitochondria. Collectively, our data demonstrate that Saussurea lappa and Pharbitis nil induce growth inhibition and apoptosis of human gastric cancer cells, and these effects are correlated with down- and up-regulation of growth-regulating apoptotic and tumor suppressor genes, respectively.

Effects of moxibustion to zusanli (ST36) on alteration of natural killer cell activity in rats.

Moxibustion is one of the major healing techniques in Oriental medicine. It has been widely used in many diseases such as rheumatoid arthritis, Hashimoto disease, breech presentation, etc. However, till now, effects of moxibustion on natural killer (NK) cell activity and relations between sympathetic nerve system (SNS) and the immune alteration induced by moxibustion were not well studied. This study was designed to evaluate effects of moxibustion on NK cell activity and the intervention of SNS in the alteration of NK cell activity induced by moxibustion. Splenic NK cell cytotoxicity was measured in a standard 4-hour 51Cr release assay. We measured the NK cell cytotoxicity after moxibustion stimulation for 1, 3, 5 and 7 days, and also measured the NK cell cytotoxicity after 3 and 7 days burn stimulation with similar temperature. Interleukin (IL)-2, -4 and interferon (INF)-gamma in serum were measured by rat IL-2, -4 and INF-gamma ELISA test kit. To evaluate the effects of sympathectomy on alteration of NK cell cytotoxicity, 6-hydroxydopamine (6-OHDA: 50 mg/kg) was used. We showed that NK cell activity of moxibustion stimulation group increased at the 3rd day, and declined at the 7th day in comparison with that of the control group. In the moxibustion stimulation group, NK cell activity was significantly higher than the sham group at the 3rd day. On the contrary, in the burn stimulation group, NK cell activity was significantly higher than that of the sham groups at 3rd and 7th days. INF-gamma level after 3 days in the moxibustion stimulation group was significantly higher than that of the sham group. IL-2 level among groups were not different. IL-4 was not detected in serum with this method. Sympathectomy abolished the NK cell activity alteration induced by moxibustion. The results suggest that moxibustion modulates NK cell activity, along with INF-gamma, and SNS is mediating these effects.