Kyoo-Seok Ahn

Deqi Sensation Between the Acupuncture-Experienced and the Naïve: A Korean Study II

Previous experience of acupuncture is believed to affect peoples expectation of future treatments. Therefore, subjects who have had acupuncture are generally excluded from sham-controlled acupuncture clinical trials. However, this assumption has not been proven, but just accepted because of the lack of evidence to the contrary. To investigate the difference in frequency and intensity of acupuncture sensation between subjects who have had acupuncture and those who have not, 36 acupuncture-experienced subjects were invited to take part in the study. After informed consent was obtained, participants were asked to complete the acupuncture sensation scale (ASS) according to what they expected needling to feel like. The needling was done at the left Hegu (LI 4) point and consisted of insertion, stimulation for 30 seconds and removal. After needling, the subjects were asked to complete the same ASS according to what they actually experienced. Adverse events were monitored. The frequency of each sensation expected and experienced, as well as acupuncture sensation scores were compared. More than 60% of the subjects expected to feel sensations of penetrating (87.6% to 100%), aching (71.2% to 95.5%), tingling (87.6% to 100%), pricking (79.7% to 99.2%) and throbbing (64.2% to 91.4%). In fact, the subjects experienced sharp (60.9% to 89.1%), intense (60.9% to 89.1%), radiating (71.2% to 95.5%) and heavy (74.8% to 97.4%) sensations just as much. The subjects expected more hurting (p = 0.001), tingling (p < 0.001), pricking (p = 0.010), stinging (p = 0.012), burning (p = 0.001) and pulsing (p = 0.009) than they experienced, while more heaviness (p = 0.011) was experienced than expected. The same outcome measures were also compared between experienced and naive groups. Apart from the fact that the acupuncture-experienced participants expected to feel pricking (p = 0.030) and stinging (p = 0.002), and experienced hurting (p = 0.022) and stinging (p = 0.028) significantly less than those who had not had acupuncture before, there was no significant difference between first time and experienced subjects. The results indicate that previous experience does not affect the peoples expectation and does not hinder people from experiencing Deqi. In addition, a constellation of Deqi-related acupuncture-specific sensations is more than just a general pain intensity dimension, which requires a biochemical and physiological exploration.

Mitochondria-cytochrome C-caspase-9 cascade mediates isorhamnetin-induced apoptosis

Isorhamnetin is a flavanoid present in plants of the Polygonaceae family and is also an immediate metabolite of quercetin in mammals. Since the plasma level of isorhamnetin is maintained longer than quercetin, isorhamnetin may be a key metabolite to mediate the anti-tumor effect of quercetin. In the present study, we investigated the apoptotic mechanism of isorhamnetin in Lewis lung cancer (LLC) cells in vitro and established its in vivo anti-cancer efficacy. In cell culture, isorhamnetin significantly increased DNA fragmentation, and TUNEL positive apoptotic bodies and sub-G(1) apoptotic population in time- and dose-dependent manners. Western blot analyses revealed increased cleavage of caspase-3, and caspase-9 and PARP and increased cytosolic cytochrome C in isorhamnetin-treated cells. These events were accompanied by a reduced mitochondrial potential. Apoptosis was blocked by a general caspase inhibitor or the specific inhibitor of caspase-3 or -9. These in vitro results support mitochondria-dependent caspase activation to mediate isorhamnetin-induced apoptosis. Furthermore, an animal study revealed for the first time that isorhamnetin given by i.p. injection at a dose that is at least one order of magnitude lower than quercetin significantly suppressed the weights of tumors excised from LLC bearing mice. The in vivo anti-tumor efficacy was accompanied by increased TUNEL-positive and cleaved-caspase-3-positive tumor cells. Our data therefore support isorhamnetin as an active anti-cancer metabolite of quercetin in part through caspase-mediated apoptosis.

Cyclooxygenase-2/prostaglandin E2 pathway mediates icariside II induced apoptosis in human PC-3 prostate cancer cells

Icariside II (IS) isolated from the roots of Epimedium koreanum Nakai was known to have antioxidant activity and inhibit melanogenesis and hypoxia inducible factor. We report here for the first time that IS induces apoptosis through its anti-inflammatory effects in PC-3 prostate cancer cells. IS exerted cytotoxicity against PC-3 cells with IC(50) of approximately 20 microM. IS suppressed both constitutive and arachidonic acid (AA)-induced cyclooxygenase-2 (COX-2) expression as well as reduced prostaglandin E2 (PGE2) levels in PC-3 cells even at a low concentrations (5 and 10 microM). Additionally, IS increased sub G1 apoptotic portion and exhibited terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL)-positive apoptotic bodies in PC-3 cells at higher concentrations (20 and 40 microM). Furthermore, IS attenuated the mitochondrial membrane potential, released cytochrome C into cytosol, activated caspase-9, -8, and -3 expressions and cleaved poly (ADP-ribose) polymerase (PARP) in PC-3 cells. Consistently, COX-2, inducible NO synthase (iNOS), and vascular endothelial growth factor (VEGF) expressions were suppressed while in parallel inducing apoptosis in hormone-independent prostate carcinoma cells PC-3. Moreover, exogeneous PGE2 inhibited IS induced PARP cleavage in PC-3 cells and also knockdown of COX-2 by siRNA potentiated IS induced PARP cleavage, thereby implicating the critical role of COX-2 pathway in IS induced apoptosis. Taken together, these findings demonstrate that IS initiates the inhibition of COX-2/PGE(2) pathway and then induces apoptosis mainly via mitochondrial dependent pathway in PC-3 prostate cancer cells as a potent cancer chemotherapeutic agent.

Effects of Korean red ginseng and its mixed prescription on the high molecular weight dextran-induced blood stasis in rats and human platelet aggregation

This study was undertaken to evaluate the antithrombotic effects of Korean Red Ginseng (KRG) and its new prescription (KRGP) consisting of five herbs such as Korean red ginseng, Ganoderma, Cinnamomi Cortex, Glycyrrhizae Radix and Laminaria. In rats with blood stasis induced by high molecular weight dextran, KRG and KRGP significantly restored not only the number of platelets and fibrinogen, but also suppressed the fibrin degradation products (FDP) to normal range. In platelet aggregation assay with human platelet rich plasma (PRP), KRG and KRGP significantly inhibited thrombin and collagen-induced platelet aggregation. The IC(50) values of KRG and KRGP were >2 and 0.23+/-0.01 mg/ml for thrombin, 0.32+/-0.01 and 0.17+/-0.02 mg/ml for collagen and 0.72+/-0.25 and >2 mg/ml for ADP, respectively. In coagulation assay, KRG and KRGP significantly prolonged activated partial prothrombin time (APPT) and prothrombin time (PT) as compared with control data. KRGP was found to be more effective than KRG alone on antithrombotic activity. These results suggest that KRGP may exert its antithrombotic activity due to inhibition of platelet aggregation and coagulation activity more than KRG.

Ocimum sanctum induces apoptosis in A549 lung cancer cells and suppresses the in vivo growth of Lewis lung carcinoma cells.

Although Ocimum sanctum has been used extensively for its medicinal values in India and China, its antitumor activity against human nonsmall cell lung carcinoma (NSCLC) A549 cells has not been investigated until now. Therefore, the antitumor mechanism of ethanol extracts of Ocimum sanctum (EEOS) was elucidated in A549 cells in vitro and the Lewis lung carcinoma (LLC) animal model. EEOS exerted cytotoxicity against A549 cells, increased the sub‐G1 population and exhibited apoptotic bodies in A549 cells. Furthermore, EEOS cleaved poly(ADP‐ribose)polymerase (PARP), released cytochrome C into cytosol and simultaneously activated caspase‐9 and ‐3 proteins. Also, EEOS increased the ratio of proapoptotic protein Bax/antiapoptotic protein Bcl‐2 and inhibited the phosphorylation of Akt and extracellular signal regulated kinase (ERK) in A549 cancer cells. In addition, it was found that EEOS can suppress the growth of LLC inoculated onto C57BL/6 mice in a dose‐dependent manner. Overall, these results demonstrate that EEOS induces apoptosis in A549 cells via a mitochondria caspase dependent pathway and inhibits the in vivo growth of LLC, suggesting that EEOS can be applied to lung carcinoma as a chemopreventive candidate. Copyright

Ethanol extract of Ocimum sanctum exerts anti-metastatic activity through inactivation of matrix metalloproteinase-9 and enhancement of anti-oxidant enzymes

Ocimum sanctum has been known to possess various beneficial properties including anti-oxidative, anti-inflammatory and anti-cancer activities. In the present study, we investigated that ethanol extracts of O. sanctum (EEOS) had anti-metastatic activity through activation of anti-oxidative enzymes. EEOS exerted cytotoxicity against Lewis lung carcinoma (LLC) cells. Also, EEOS significantly inhibited cell adhesion and invasion as well as activities of matrix metalloproteinase-9 (MMP-9), but not MMP-2, indicating the important role of MMP-9 in anti-metastatic regulation of EEOS. In addition, EEOS significantly reduced the tumor nodule formation and lung weight in LLC-injected mice. Inhibitory effect of EEOS on metastasis was further confirmed by using hematoxylin and eosin (H&E) staining. Notably, we also found that EEOS enhanced activities of anti-oxidative enzymes such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) in a concentration-dependent manner. Taken together, our findings support that EEOS can be a potent anti-metastatic candidate through inactivation of MMP-9 and enhancement of anti-oxidant enzymes.

DMNQ S-64 Induces Apoptosis via Caspase Activation and Cyclooxygenase-2 Inhibition in Human Nonsmall Lung Cancer Cells

Abstract:u2002 Shikonin has been reported to induce apoptosis and inhibit angiogenesis in vivo and in vitro. 6‐(1‐propoxyiminoalkyl)‐5,8‐dimethoxyoxy 1,4‐naphtoquinone S‐64 (DMNQ S‐64) was synthesized as a shikonin derivative. In this article, the underlying apoptotic mechanism of DMNQ S‐64 was examined. DMNQ S‐64 exerted cytotoxicity against A549 lung carcinoma cells with IC50 of 27.3 μM. Apoptotic bodies were observed in DMNQ S‐64‐treated A549 cells by 4′‐6‐diamidino‐2‐phenylindole (DAPI) staining assay. DMNQ S‐64 also increased sub‐G1 DNA portion in a concentration‐dependent manner by flow cytometric analysis. Western blotting has revealed that DMNQ S‐64 effectively activates the expression of caspase 8, 9, and 3, cleaves poly (ADP‐ribose) polymerase, and increases the ratio of Bax/Bcl‐2. Furthermore, cytochrome c was released in a concentration‐dependent manner by DMNQ S‐64. Similarly, DMNQ S‐64 significantly increased caspase 3 activity by enzyme‐linked immunosorbent assay (ELISA). It also significantly inhibited the level of prostaglandin E2 (PGE2) by ELISA and downregulated the expression of cyclooxygenase‐2 (COX‐2) in a concentration‐dependent manner. Taken together, DMNQ S‐64 may exhibit cytotoxicity against A549 cells via caspase activation and COX‐2 inhibition.

Inhibitory effect of Platycodon grandiflorum on TH1 and TH2 immune responses in a murine model of 2,4-dinitrofluorobenzene–induced atopic dermatitis–like skin lesions

BACKGROUNDnPlatycodon grandiflorum is a traditional Asian medicine that is used to treat pulmonary and respiratory allergic disorders.nnnOBJECTIVEnto investigate the effects of P grandiflorum in vivo in an animal model of atopic dermatitis (AD), with particular emphasis on its effects on T(H)1 and T(H)2 immune responses.nnnMETHODSnwe established a model of AD-like skin lesions in NC/Nga mice. After oral administration of P grandiflorum, we measured cytokine and immunoglobulin profiles along with histologic examination of skin.nnnRESULTSnP grandiflorum was nontoxic in a 2,4-dinitrofluorobenzene-induced model of AD-like skin lesions in NC/Nga mice. AD symptoms in skin lesions improved after oral administration of P grandiflorum. IgE secretion was significantly downregulated in P grandiflorum-treated animals, accompanied by decreased levels of interleukin (IL) 4 and IgG1 and increased serum levels of IL-12p40 and IgG2a. In isolated splenocytes, the production of the T(H)1 cytokines IL-12p40 and interferon-γ was upregulated by P grandiflorum, whereas the levels of the T(H)2 cytokines IL-4 and IL-5 were downregulated in a mouse model of AD-like skin lesions.nnnCONCLUSIONSnthese results suggest that P grandiflorum inhibits the development of AD-like skin lesions in NC/Nga mice by suppressing the T(H)2 cell response and increasing the T(H)1 cell responses. Our results indicate that P grandiflorum is safe and effective as a natural herbal medicine for the treatment of AD-like skin lesions.

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)‐γ 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‐γ 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.

Uncoupled protein 3 and p38 signal pathways are involved in anti-obesity activity of Solanum tuberosum L. cv. Bora Valley

AIM OF STUDYnThis study was undertaken to elucidate the anti-obesity mechanism of a new purple potato variety that has been used for the prevention of metabolic diseases as a folk remedy in Korea.nnnMATERIALS AND METHODSnProliferation assay, differentiation assay, Western blotting, were performed in 3T3-L1 adipocytes, while blood chemistry for hyperlipidemic parameters, measurement of body weight and abdominal fats, reverse transcriptase-polymerase chain reaction (RT-PCR) analysis, magnetic resonance image (MRI) scanning were carried out in high fat diet fed Sprague-Dawley rats with ethanol extract of Solanum tuberosum L. cv. Bora Valley (ESTBV).nnnRESULTSnESTBV significantly inhibited the proliferation and differentiation of 3T3-L1 cells as well as reduced the cellular leptin level. ESTBV also significantly attenuated the levels of insulin and leptin at 500mg/kg in high fat diet fed rats. In addition, ESTBV significantly reduced total fat and whole body lipid in a therapeutic experiment, which was confirmed by MRI scanning and also significantly inhibited the retroperitoneal and epididymal fats in a preventive experiment compared with control. Similarly, the levels of total cholesterol, triglyceride and low density lipoprotein (LDL) were significantly reduced at a lower dose 200mg/kg of ESTBV in a preventive experiment than at 500mg/kg in a therapeutic experiment. Furthermore, body weight gain was significantly suppressed by over 4 weeks treatment of ESTBV compared with control. Interestingly, the expression of p38 mitogen-activated protein kinase (MAPK) was significantly downregulated in 3T3-L1 cells by ESTBV and the expression of uncoupled protein 3 (UCP-3) was activated in fats and liver tissues of ESTBV treated group compared with high fat diet control.nnnCONCLUSIONnESTBV has anti-obesity potential via inhibition of lipid metabolism through p38 MAPK and UCP-3 pathways.