Objectives : Alzheimer's disease (AD) is a geriatric dementia that is widespread in old age. In the near future AD will be the biggest problem in public health service. Although a variety of oriental prescriptions, including Sesim-tang, have been traditionally utilized for the treatment of AD, their pharmacological effects and action mechanisms have not yet been fully elucidated. The present study investigated the effects of Sesim-tang on apoptotic cell death induced by CT105 (carboxy terminal 105 amino acid peptide fragment of APP) overexpression in SK-N-SH neuroblastoma cell lines. Methods: We studied the regenerative and inhibitory effects on Alzheimer's disease in CT105-induced SK-N-SH cell lines by Sesim-tang water extract. We examined for cell morphological pattern, DNA fragmentation, LDH activity assay, zymography assay, and immunohistochemistric analysis. Additionally, we investigated the association between the CT105 and neurite degeneration caused by CT105-induced apoptotic response in neurone cells. Results: Findings from our experiments have shown that Sesim-tang inhibits the synthesis or activities of CT105, which has neurotoxicities and apoptotic activities in the cell line. In addition, pretreatment with Sesim-tang ($>50\mu\textrm{g}/ml$ for 12 hours) partially prevented CT105-induced cytotoxicity in SK-N-SH cell lines. SK-N-SH cell lines overexpressed with CT105 exhibited remarkable apoptotic cell damage. Based on morphological observations by phase-contrast microscope and LDH activity measurements in the culture media, the CT105-induced cell death was significantly inhibited by Sesim-tang water extract. Sesim-tang was found to reduce the expression of APP and caspase-3 induced by CT105 in SK-N-SH cell lines and in rat hippocampus. Conclusions: As the result of this study, in the Sesim-tang group, apoptosis in the nervous system is inhibited, the repair against the degeneration of SK-N-SH cell lines by CT105 expression is promoted. Hence, Sesim-tang may be beneficial for the treatment of AD.
Potassium channels in human skin fibroblast have been studied as a possible site of Alzheimer disease pathogenesis. Fibroblasts in Alzheimer disease show alterations in signal transduction pathway such as changes in $Ca^{2+}$ homeostasis and/or $Ca^{2+}-activated$ kinases, phosphatidylinositol cascade, protein kinase C activity, cAMP levels and absence of specific $K^+$ channel. However, little is known so far about electrophysiological and pharmacological characteristics of large-conductance $Ca^{2+}$-activated $K^+$ ($BK_{Ca}$) channel in human fibroblast (CRL-1474). In the present study, we found Iberiotoxin- and TEA-sensitive outward rectifying oscillatory current with whole-cell recordings. Single channel analysis showed large conductance $K^{+}$ channels (106 pS of chord conductance at +40 mV in physiological $K^+$ gradient). The 106 pS channels were activated by membrane potential and $[Ca^{2+}]_i$, consistent with the known properties of $BK_{Ca}$ channels. $BK_{Ca}$ channels in CRL-1474 were positively regulated by adenylate cyclase activator ($10{\mu}M$ forskolin), 8-Br-cyclic AMP ($300{\mu}M$) or 8-Br-cyclic GMP ($300{\mu}M$). These results suggest that human skin fibroblasts (CR-1474) have typical $BK_{Ca}$ channel and this channel could be modulated by c-AMP and c-GMP. The electrophysiological characteristics of fibroblasts might be used as the diagnostic clues for Alzheimer disease.
Kim Chang Hwan;Kim Han Geu;Ahan Jong Chan;Lee Soo Kyung;Chung Tae Wook;Kim June Ki;Choi Dall Yeong;Kim Cherl Ho;Park Won Hwan
동의생리병리학회지
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제16권6호
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pp.1277-1283
/
2002
Silsosangami(SSG) is a formula of treaditional korean medicines as an effective biological response modifier for augmenting host homeostasis of body circulation. Little is known of the biological activity of SSG and previous studies have focused mainly on their anti-thrombosis8). There is a growing interest in the pharmacological potential of the SSG due to the recent finding by our group that SSG and each constituent herbs of SSG were able to inhibit NO and prostaglandin E2 (PGE2) synthesis in murine peritoneal macrophages stimulated with bacterial endotoxin. In this paper, the effects of SSG on platelet aggregation and hemolysis in human blood were studied. SSG provoked remarkable inhibiting effect on platelet aggregation, and APTT were sensitive to the presence of this SSG. Using an in vitro system, APTT was delayed with the increment of the concentrations of these seven compounds. These results suggested that SSG might be used as a novel antithrombotic therapeutic agents in post-myocardial infarction. A SSG reduced NO production in mouse peritoneal macrophages stimulated with lipopolysaccharide, without the influence on the activity of iNOS being observed. SSG significantly reduced mouse paw edema induced by carrageenan. Western blot analysis showed that SSG reduced the expression of iNOS. The results indicate that SSG exerts anti-inflammatory effects related to the inhibition of NO production, which could be due to a decreased expression of iNOS.
Imperata cylindrica, a tall tufted grass which has multiple pharmacological applications is one of the key ingredients in various traditional medicinal formula used in India. Previous reports have shown that I. cylindrica plant extract inhibited cell proliferation and induced apoptosis in various cancer cell lines. To our knowledge, no studies have been published on the effect of I. cylindrica leaf extract on human oral cancers. The present study was undertaken in order to evaluate the anticancer properties of the leaf extract of I. cylindrica using an oral squamous cell carcinoma cell line SCC-9 as an in vitro model system. A methanol extract from dried leaves of I. cylindrica (ICL) was prepared by standard procedures. Effects of the ICL extract on the morphology of SCC-9 cells was visualized by microscopy. Cytotoxicity was determined by MTT assay. Effects of the ICL extract on colony forming ability of SCC-9 cells was evaluated using clonogenic assay. Cell cycle analysis was performed by flow cytometry and induction of apoptosis was determined by DNA fragmentation assay. The ICL extract treatment caused cytotoxicity and induced cell death in vitro in SCC-9 cells in a dose-dependent manner. This treatment also significantly reduced the clonogenic potential and inhibited cell proliferation by arresting the cell cycle in the G2/M phase. Furthermore, DNA fragmentation assays showed that the observed cell death was caused by apoptosis. This is the first report showing the anticancer activity of the methanol extracts from the leaves of I. cylindrica in human oral cancer cell line. Our data indicates that ICL extract could be considered as one of the lead compounds for the formulation of anticancer therapeutic agents to treat/manage human oral cancers. The natural abundance of I. cylindrica and its wide geographic distribution could render it one of the primary resource materials for preparation of anticancer therapeutic agents.
In this double-blind, placebo-controlled trial, we assessed the efficacy and safety of beef tallow extract (BTE) including Cis-9-cetylmyristoleate in patients with arthritis. Between May and December 2003, we selected 80 patients (n=80) who showed/manifested arthritic symptoms and whose radiological findings were suggestive of arthritis, and randomly assigned them to placebo-controlled (n=40) and treatment group (n=40). The placebo (corn starch 350 mg) and BTE (208 mg) were orally administered to placebo-controlled and treatment group three times a day, respectively. We assessed the efficacy and safety based on the visual analogue scale (V AS) and modified knee society knee scores (MKSKS) at baseline and endpoint, respectively. To assess the safety, we monitored the adverse effects noted in liver, kidney, cardiovascular and gastrointestinal system for 3 weeks. Then, we performed not only a questionnaire study but also laboratory tests (e.g., liver function test, kidney function test, urinalysis, electrocardiography [EKG], complete blood cell counts [CBC] and chest X-ray). For statistical analysis, Student (-test and paired (-test were done using SPSS■ Version 11.0. Statistical significance was set at p < 0.05. The scores between V AS and MKSKS showed statistical significance (p < 0.05) with an improvement of $69.2\%$ (27/39) and $3.8\%$ (21/39) of treatment-group patients, respectively. Abnormal laboratory findings were noted in neither placebo-controlled nor treatment group. In conclusion, our results indicate that the administration of BTE was a safe and effective treatment regimen for patients with arthritis. In addition, the efficacy of BTE was more remarkable in alleviating the symptoms rather than improving the function.
Background: Panax ginseng has been used for a variety of medical purposes in eastern countries for more than two thousand years. From the extensive experiences accumulated in its long medication use history and the substantial strong evidence in modern research studies, we know that ginseng has various pharmacological activities, such as antitumor, antidiabetic, antioxidant, and cardiovascular system-protective effects. The active chemical constituents of ginseng, ginsenosides, are rich in structural diversity and exhibit a wide range of biological activities. Methods: Ginsenoside constituents from P. ginseng flower buds were isolated and purified by various chromatographic methods, and their structures were identified by spectroscopic analysis and comparison with the reported data. The 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H- tetrazolium bromide method was used to test their cytotoxic effects on three human cancer cell lines. Results: Six ginsenosides, namely 6'-malonyl formyl ginsenoside F1 (1), 3β-acetoxyl ginsenoside F1 (2), ginsenoside Rh24 (6), ginsenoside Rh25 (7), 7β-hydroxyl ginsenoside Rd (8) and ginsenoside Rh26 (10) were isolated and elucidated as new compounds, together with four known compounds (3-5 and 9). In addition, the cytotoxicity of these isolated compounds was shown as half inhibitory concentration values, a tentative structure-activity relationship was also discussed based on the results of our bioassay. Conclusion: The study of chemical constituents was useful for the quality control of P. ginseng flower buds. The study on antitumor activities showed that new Compound 1 exhibited moderate cytotoxic activities against HL-60, MGC80-3 and Hep-G2 with half inhibitory concentration values of 16.74, 29.51 and 20.48 μM, respectively.
Kim, Seong-Il;Raffi Mikaelian;Kwak, Jin-Hwan;Kim, In-Chull;Lee, Chang-Ho
Biomolecules & Therapeutics
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제3권4호
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pp.316-321
/
1995
All the pharmacological studies of LB17522 described here were carried out with high doses (fifteen to sixty times of the therapeutic dose) to determine an indication of potential side effects in clinical use in terms of the acute clinical signs, cardiovascular and central nervous system. LB10522 does not produce any observable clinical signs except for the symptoms such as moist eye, skin rash, slight salivation, vomitting, and slightly reduced activity. The effects of LB10522 on the hemodynamics and cardiac function of anesthetized beagle dogs are as follows; heart rates and mean arterial blood pressure had a tendency to increase mildly, which is a normal finding in anesthetized dogs. All the animals except for one showed relatively stable respiratory rates throughout the observation period. Each animal treated with LB10522 showed slight increase in the left cardiac work and left ventricular stroke work which are mainly related to corresponding increases in cardiac output. Femoral blood flow were shown to be increased in some animals treated with LB10522. The epileptogenic activities of various cephalosporins were assessed by a direct intracerebral injection of appropriate concentration of test articles. The CD$_{50}$ values (nmol) obtained from the analysis of the dose-response data are as follows; 78.2, 175.3, 156.3, and 53.5 for cefazolin, cephaloridine, ceftazidime, and LB 10522, respectively. LB10522 seems to be equipotent with cefazolin or to be three times more potent than cephaloridine and ceftazidime in causing adverse CNS stimulation. Taken into consideration all the information obtained, LB10522 is not supposed to induce much changes in the functions examined in these studies in man at therapeutic doses.s.
Methamphetamine (MA) is currently the most abused illicit drug in Korea and its major metabolite is amphetamine (AP). As MA exist as two enantiomers with the different pharmacological properties, it is necessary to determine their respective amounts in a sample. Thus a chiral stationary phase liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed for identification and quantification of d-MA, l-MA, d-AP, and l-AP in human urine. Urine sample ($200{\mu}L$) was diluted with pure water and purified using solid-phase extraction (SPE) cartridge. A $5-{\mu}L$ aliquot of SPE treated sample solution was injected into LC-MS/MS system. Chiral separation was carried out on the Astec Chirobiotic V2 column with an isocratic elution for each enantiomer. Identification and quantification of enantiomeric MA and AP was performed using multiple reaction monitoring (MRM) detection mode. Linear regression with a $1/x^2$ as the weighting factor was applied to generate a calibration curve. The linear ranges were 25-1000 ng/mL for all compounds. The intra- and inter-day precisions were within 3.6 %, while the intra- and inter-day accuracies ranged from -5.4 % to 11.8 %. The limits of detection were 2.5 ng/mL (d-MA), 3.5 ng/mL (l-MA), 7.5 ng/mL (d-AP), and 7.5 ng/mL (l-AP). Method validation parameters such as selectivity, matrix effect, and stability were evaluated and met acceptance criteria. The applicability of the method was tested by the analysis of genuine forensic urine samples from drug abusers. d-MA is the most common compound found in urine and mainly used by abusers.
Jawarishe Jalinoos (JJ) is an orally used formulation available in semisolid dosage form, prepared with powdered plant materials mixed in honey or sugar syrup. It has many admirable pharmacological effects and used in Unani medicine to treat various acute and chronic disorders since ancient times. The ICH Harmonised Tripartite Guideline stated that photostability testing should be an essential part of stability testing to confirm that light exposure does not result in an unacceptable change in drugs substance and finished products. To date, the effect of light on JJ is not studied, in this study photostability evaluation of JJ was carried out. The test sample was manufactured with genuine ingredients in the in-door pharmacy of the National Institute of Unani Medicine. JJ was packed in two transparent polyethylene terephthalate airtight containers. The first sample was analysed at zero-day and the second sample was placed in a stability chamber subjected to light challenge with an overall illumination of 1.2 million lux hours combined with near ultraviolet energy of 200-watt hours per square meter by using option 2, along with 30±2℃ temperature and relative humidity 70±5%. Analysis of both finished products showed no considerable changes in organoleptic characters. Less than 5% variation was observed in physicochemical parameters. HPTLC fingerprinting showed justifiable variation. Microbial load and specific counts were within the limit prescribed by WHO. As no unacceptable changes were noted in JJ subjecting to light challenge, it is concluded that JJ is a photostable Unani compound formulation.
Results of an experiment on the behavior of rats and mice in order to explore the possible pharmacological actions of Panax ginseng upon the central nervous system can be summarized as follows: 1. Spontaneous motor activity. In the case of mice, those groups who were administered 2.5 mg and 5.0mg of ginseng saponin per kilogram of body weight were observed to have increased their activity compared with the control group, while the 50.0 mg and 100.0 mg per kilogram body weight groups demonstrated lower levels of activity, with the peak of activity appearing at 30 minutes after administration of drugs. In the case of rats, those groups of animals who were given injections in the dosage of 2.5 mg, 5.0 mg and 50.0 mg per kilogram body weight demonstrated higher activity than the control group, while the 100.0 mg per kilogram group appeared to have decreased in their activity, with the peak action appearing 30 minutes after the administration of ginseng saponin. The 50.0 mg per kilogram group demonstrated no significant differential. 2. General behavior analysis. In the case of mice, decrease in sleeping component of behavior and increase in the walking and roaring components, compared those with the control group, turned out to be a common phenomenon among the groups who were administered 2.5 mg, 5.0 mg and 50.0 mg of ginseng saponin per kilogram body weight, with the 5.0 mg per kilogram group standing out of all the other groups in terms of their reactions. In the case of rats, ginseng saponin appeared to reduce sleeping component with 2.5 mg, 5.0 mg and 50.0 mg per kilogram body weight groups, while increased the walking and rearing components. It was observed that administratoin of ginseng saponin in a dose of 2.5 mg per kilogram appeared to markedly increase the lying and grooming components of animal behavior. 3. Open-field exploratory behavior. Adminstration of ginseng saponin to mice in doses of 5.0 mg, 50.0 mg and 100.0 mg per kilogram body weight decreased activity, but increased their exploratory behavior. In the case of rats, however, administration of ginseng saponin in the doses of 2.5 mg and 5.0 mg per kilogram body weight markedly increased their activities, while decreased activities with the 50.0 mg per kilogram and 100.0 mg per kilogram groups. The exploratory behavior of rats appeared to have decreased, while grooming increased ramarkably. 4. The above findings from a series of experiment appear to suggest a stimulating effect on the central nervous system when ginseng saponin is administered in small doses, but that larger doses might result in an inhibitory effect, though differential results can be anticipated with modification of experimental conditions.
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