• Title/Summary/Keyword: In vivo

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Phytochemical Analysis and Anti-cancer Investigation of Boswellia Serrata Bioactive Constituents In Vitro

  • Ahmed, Hanaa H;Abd-Rabou, Ahmed A;Hassan, Amal Z;Kotob, Soheir E
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7179-7188
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    • 2015
  • Cancer is a major health obstacle around the world, with hepatocellular carcinoma (HCC) and colorectal cancer (CRC) as major causes of morbidity and mortality. Nowadays, there isgrowing interest in the therapeutic use of natural products for HCC and CRC, owing to the anticancer activity of their bioactive constituents. Boswellia serrata oleo gum resin has long been used in Ayurvedic and traditional Chinese medicine to alleviate a variety of health problems such as inflammatory and arthritic diseases. The current study aimed to identify and explore the in vitro anticancer effect of B. Serrata bioactive constituents on HepG2 and HCT 116 cell lines. Phytochemical analysis of volatile oils of B. Serrata oleo gum resin was carried out using gas chromatography-mass spectrometry (GC/MS). Oleo-gum-resin of B. Serrata was then successively extracted with petroleum ether (extract 1) and methanol (extract 2). Gas-liquid chromatography (GLC) analysis of the lipoidal matter was also performed. In addition, a methanol extract of B. Serrata oleo gum resin was phytochemically studied using column chromatography (CC) and thin layer chromatography (TLC) to obtain four fractions (I, II, III and IV). Sephadex columns were used to isolate ${\beta}$-boswellic acid and identification of the pure compound was done using UV, mass spectra, $^1H$ NMR and $^{13}C$ NMR analysis. Total extracts, fractions and volatile oils of B. Serrata oleo-gum resin were subsequently applied to HCC cells (HepG2 cell line) and CRC cells (HCT 116 cell line) to assess their cytotoxic effects. GLC analysis of the lipoidal matter resulted in identification of tricosane (75.32%) as a major compound with the presence of cholesterol, stigmasterol and ${\beta}$-sitosterol. Twenty two fatty acids were identified of which saturated fatty acids represented 25.6% and unsaturated fatty acids 74.4% of the total saponifiable fraction. GC/MS analysis of three chromatographic fractions (I,II and III) of B. Serrata oleo gum resin revealed the presence of pent-2-ene-1,4-dione, 2-methyl- levulinic acid methyl ester, 3,5- dimethyl- 1-hexane, methyl-1-methylpentadecanoate, 1,1- dimethoxy cyclohexane, 1-methoxy-4-(1-propenyl)benzene and 17a-hydroxy-17a-cyano, preg-4-en-3-one. GC/MS analysis of volatile oils of B. Serrata oleo gum resin revealed the presence of sabinene (19.11%), terpinen-4-ol (14.64%) and terpinyl acetate (13.01%) as major constituents. The anti-cancer effect of two extracts (1 and 2) and four fractions (I, II, III and IV) as well as volatile oils of B. Serrata oleo gum resin on HepG2 and HCT 116 cell lines was investigated using SRB assay. Regarding HepG2 cell line, extracts 1 and 2 elicited the most pronounced cytotoxic activity with $IC_{50}$ values equal 1.58 and $5.82{\mu}g/mL$ at 48 h, respectively which were comparable to doxorubicin with an $IC_{50}$ equal $4.68{\mu}g/mL$ at 48 h. With respect to HCT 116 cells, extracts 1 and 2 exhibited the most obvious cytotoxic effect; with $IC_{50}$ values equal 0.12 and $6.59{\mu}g/mL$ at 48 h, respectively which were comparable to 5-fluorouracil with an $IC_{50}$ equal $3.43{\mu}g/mL$ at 48 h. In conclusion, total extracts, fractions and volatile oils of B. Serrata oleo gum resin proved their usefulness as cytotoxic mediators against HepG2 and HCT 116 cell lines with different potentiality (extracts > fractions > volatile oil). In the two studied cell lines the cytotoxic acivity of each of extract 1 and 2 was comparable to doxorubicin and 5-fluorouracil, respectively. Extensive in vivo research is warranted to explore the precise molecular mechanisms of these bioactive natural products in cytotoxicity against HCC and CRC cells.

Effects of Repeated Superovulation on Recovery and Quality of Oocytes in Korean Native Goats (재래산양의 반복 과배란처리에 의한 난자의 회수율과 등급에 관한 연구)

  • Park, H.-S.;Lee, Y.-H.;Kim, T.-S.;Park, J.-K.;Lee, J.-S.;Kim, C.-H.;Jung, J.-Y.
    • Journal of Embryo Transfer
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    • v.19 no.2
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    • pp.81-87
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    • 2004
  • This study was designed to determine whether repeated superovulation is beneficial for recovery and quality of oocytes in Korean native goats. Seventy-six mature goats, maintained in a pen under natural day length and fed hay ad libitum, were pretreated with progestagen impregnated CIDR for 10 days and then the goats were divided into two groups. One group of the goats received a single intramuscular injection of 1,000 IU PMSG on Day 8 of CIDR insertion. The other group of the goats received twice daily intramuscular injections of a total of 70 mg FSH for 3 days from Day 8 of CIDR. All the gonadotropin treated goats were injected with 10 mg $PGF2{\alpha}on$ Day 8 and 400 IU hCG in the afternoon on Day 10. For oocyte recovery, donor goats were fasted 24 h before operation. Anesthesia was induced by intravenous injection of 2% xylazine(0.2 mg/kg body weight) and ketamin(11 mg/kg body weight). In vivo oocytes were recovered by follicle aspiration or oviduct flushing at 35 to 40 hours after hCG injection through mid-ventral incision. The mean number of CL and oocytes recovered and recovery rate of oocytes by oviduct flushing were greater(P<0.05) in the first treatment than those in the second treatment. Contrary to our assumption, PMSG treatment significantly (P<0.05) increased the number of CL formed and recovery rate of oocytes compared to FSH. However, the same effect was not observed in recovery of follicular oocytes. There was no significant difference in oocyte quality between FSH and PMSG or first and second treatments. The present results indicate that repeated superovulation and repeated use of donor animals may be inefficient for obtaining oocytes in good qualities.

The Food Safety of Superfine Powder (Phellinus linteus) Processed by Nanomill in C57BL/6 Mice (C57BL/6 마우스에서 나노밀 가공된 초미세분말(상황버섯)의 식이 안전성 연구)

  • Kim, Dong-Heui;Teng, Yung-Chien;Yoon, Yang-Sook;Qi, Xu-Feng;Jeong, Hyun-Seok;Joo, Kyung-Bok;Lee, Kyu-Jae
    • Applied Microscopy
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    • v.39 no.2
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    • pp.133-139
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    • 2009
  • A officinal mushroom, Phellinus linteus (PL) has been known to exhibit potent biological activities including antioxidative and anticancer effect. PL is consumed as a type of powder or extract for the purpose of health promotion and disease treatment. Recently superfine PL products was commercialized according to the development of pulverizing technology such as nanomill, so the evaluation of food safety is suggested. This study was conducted to evaluate the food safety of superfine PL (SPL) through hematological, biochemical and histological examination in mice as compared with fine PL (FPL). In the particle size distribution in volume after nanomill processing, the mean diameter of SPL and FPL particles was 11.78 ${\mu}m$ and 216.1 ${\mu}m$, and d (0.5), the particle diameter measured at 50% of distribution was 5.5 ${\mu}m$ and 147.9 ${\mu}m$, respectively. As the result of body weight, food intake and the weight of organs, SPL group didn't show any statistical difference compared with FPL group and normal group (N). Hematological and biochemical values were also involved in the normal range, although ALT (N vs. FPL, P<0.001) and BUN (N vs. FPL, P<0.01; N vs. SPL, P<0.01) showed significance compared with N group but there are no significance between FPL and SPL group. In the result of histological examination with liver, kidney, spleen, and small and large intestine, abnormal findings such as inflammatory reaction and histological changes were not observed. Our results suggest that the oral intake of SPL diet is not harmful to the animal in the hematological, biochemical and histological aspects although particle size was reduced to the level of superfine. However, further study will be necessary to confirm the histological safety in relation to the gastrointestinal contact of superfine particles in the case of large amount and long-term intake.

Induction of Apoptosis and Antitumor Activity by Stichoposide D through the Generation of Ceramide in Human Leukemia Cells (Stichoposide D의 백혈병 세포주에서 세라마이드 생성을 통한 세포 사멸 유도 및 항암 작용)

  • Park, Eun-Seon;Yun, Seung-Hoon;Shin, Sung-Won;Kwak, Jong-Young;Park, Joo-In
    • Journal of Life Science
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    • v.22 no.6
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    • pp.760-771
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    • 2012
  • Marine triterpene glycosides are physiologically active natural compounds isolated from sea cucumbers(holothurians). It was demonstrated that they have a wide range of biological activities, including antifungal, cytotoxic, and antitumor effects. A previous study showed that stichoposide C (STC) isolated from Thelenota anax induces apoptosis through generation of ceramide by activation of acid sphingomyelinase (SMase) and neutral SMase in human leukemia cells. In this study, we investigated whether STD, a structural analog of STC, can induce apoptosis and examined the molecular mechanisms for its activity. It was found that STC and STD induce apoptosis in a dose- and time-dependent manner and lead to the activation of caspase-8, mitochondrial damage, activation of caspase-9, and activation of caspase-3 in K562 and HL-60 cells. STC activates acid SMase and neutral SMase, which results in the generation of ceramide. Specific inhibition of acid SMase or neutral SMase partially blocked STC-induced apoptosis, but not STD-induced apoptosis. In contrast, STD generates ceramide through the activation of ceramide synthase. Specific inhibition of ceramide synthase partially blocked STD-induced apoptosis, but not STC-induced apoptosis. Moreover, STC and STD markedly reduced tumor growth of HL-60 xenograft tumors and increased ceramide generation in vivo. These results indicate that STC and STD can induce apoptosis and have antitumor activity through the different molecular mechanisms, because they have a different sugar residue attached to aglycones. Thus, these results suggest that their actions are affected by a sugar residue attached to aglycones and they can be used as anticancer agents in the treatment of leukemia.

Effect of Buan Mulberry on Metabolic Improvement in Streptozotocin-Induced Diabetic Rats (Streptozotocin으로 유발된 당뇨쥐의 신진대사 증진에 대한 부안 오디의 효과)

  • Lee, Ju-Taek;Ryu, Jae-Sung;Kwak, Dong-Hoon;Park, Yun-Jum;Kang, Seong-Sun;Kim, Pyoung-Jun;Hwang, Kyung-A;Choo, Young-Kug
    • Journal of Life Science
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    • v.20 no.12
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    • pp.1750-1757
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    • 2010
  • This study was designed to evaluate the anti-diabetic effects of Buan mulberries by using an insulin-dependent diabetes mellitus animal model. Several studies have shown that mulberries have metabolism-improving, antioxidant, and lipid-lowering properties in rats with streptozotocin (STZ)-induced diabetes. In this study, Sprague-Dawley male rats were randomly assigned to 1 normal control group and 5 STZ-induced diabetes groups: rats that had STZ-induced diabetes and did not receive any agents (diabetic group; negative control), rats that had STZ-induced diabetes and received insulin (insulin group; positive control), rats that had STZ-induced diabetes and received 0.5% mulberry extract (0.5% mulberry group), rats that had STZ-induced diabetes and received 1.0% mulberry extract (1.0% mulberry group), and rats that had STZ-induced diabetes and received 2.0% mulberry extract (2.0% mulberry group). Mulberry extracts were administered to the diabetic animals for 4 weeks. The rats that received mulberry extracts showed lower body weights and insulin levels, as well as higher kidney weights, blood glucose levels, urine quantities, and water intake in comparison with the normal controls. Further, the insulin concentrations in the mulberry-fed animals were higher than those in the diabetic group, and the kidney weights, blood glucose levels, urine quantities, and water intake in the mulberry-fed animals were lower than the corresponding values in the diabetic controls. These results suggest that mulberry may be an effective functional food to prevent diabetes-related complications.

Regrowth Ability and Species Composition of Phytoplankton in International Commercial Ship's Ballast Water Berthed at Pusan and Daesan Ports (부산과 대산항에서 선박평형수에 유입된 식물플랑크톤의 종조성과 재성장능력)

  • Baek, Seung-Ho;Jang, Min-Chul;Shin, Kyoung-Soon
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.16 no.2
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    • pp.106-115
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    • 2011
  • The aim of this study is to assess the importance of ballast water discharge as a vector for the introduction of exotic species into Pusan and Daesan Ports, Korea. We also examined to understand the impacts of environmental factors on the survival success of introduced species by ship's ballast water in laboratory experiments. Seven ship's ballast water originated from the coastal water of China (Taicang, Ningbo and Jinshan), Japan (Tokuyama, Moji and Akita), and Singapore. According to PCA (principal components analysis) analysis, environmental factor in the each ballast and shipside waters were different by bioregion. Based on cluster analysis, the phytoplankton community structures were distinguished for ballast water origin. Most of the major taxonomic groups were diatoms and, the others were dinoflagellate, silcoflagellate and several fresh-waters species. In particular, species number and standing crops of phytoplankton in the ballast tanks decreased with the increasing age ofballast water(r = -0.35 for standing crop; r = -0.63 for species number). In the laboratory study, although phytoplankton in ballast water treatment did not survive even in optimal temperature, the in vivo fluorescence of phytoplankton viability increased under the nutrient typical of shipside water and F/2 medium at $15^{\circ}C$ and $20^{\circ}C$. The diatoms species such as Skeletonema costatum and Thalassiosira pseudonana in ballast water were successfully regrown. On the salinity gradient experiments for Shui Shan (2) vessel, several freshwater species, brackish and marine species were successfully adapted. Of these, S.costatum was able to tolerate a wide range of salinities (10 to 30 psu) and its species-specific viability was suitable for colonization.

Radiation Absorbed Dose Calculation Using Planar Images after Ho-166-CHICO Therapy (Ho-166-CHICO 치료 후 평면 영상을 이용한 방사선 흡수선량의 계산)

  • 조철우;박찬희;원재환;왕희정;김영미;박경배;이병기
    • Progress in Medical Physics
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    • v.9 no.3
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    • pp.155-162
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    • 1998
  • Ho-l66 was produced by neutron reaction in a reactor at the Korea Atomic Energy Institute (Taejon, Korea). Ho-l66 emits a high energy beta particles with a maximum energy of 1.85 MeV and small proportion of gamma rays (80 keV). Therefore, the radiation absorbed dose estimation could be based on the in-vivo quantification of the activity in tumors from the gamma camera images. Approximately 1 mCi of Ho-l66 in solution was mixed into the flood phantom and planar scintigraphic images were acquired with and without patient interposed between the phantom and scintillation camera. Transmission factor over an area of interest was calculated from the ratio of counts in selected regions of the two images described above. A dual-head gamma camera(Multispect2, Siemens, Hoffman Estates, IL, USA) equipped with medium energy collimators was utilized for imaging(80 keV${\pm}$10%). Fifty-nine year old female patient with hepatoma was enrolled into the therapeutic protocol after the informed consent obtained. Thirty millicuries(110MBq) of Ho-166-CHICO was injected into the right hepatic arterial branch supplying hepatoma. When the injection was completed, anterior and posterior scintigraphic views of the chest and pelvic regions were obtained for 3 successive days. Regions of interest (ROIs) were drawn over the organs in both the anterior and posterior views. The activity in those ROIs was estimated from geometric mean, calibration factor and transmission factors. Absorbed dose was calculated using the Marinelli formula and Medical Internal Radiation Dose (MIRD) schema. Tumor dose of the patient treated with 1110 MBq(30 mCi) Ho-l66 was calculated to be 179.7 Gy. Dose distribution to normal liver, spleen, lung and bone was 9.1, 10.3, 3.9, 5.0 % of the tumor dose respectively. In conclusion, tumor dose and absorbed dose to surrounding structures were calculated by daily external imaging after the Ho-l66 therapy for hepatoma. In order to limit the thresholding dose to each surrounding organ, absorbed dose calculation provides useful information.

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The analysis of ethylene glycol and metabolites in biological specimens (생체시료에서 에틸렌 글리콜과 그 대사체 분석에 관한 연구)

  • Park, Seh-Youn;Kim, Yu-Na;Kim, Nam-Yee
    • Analytical Science and Technology
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    • v.24 no.2
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    • pp.69-77
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    • 2011
  • Ethylene glycol (EG) is produced commercially in large amounts and is widely used as antifreeze or deicing solution for cars, boats, and aircraft. EG poisoning occurs in suicide attempts and infrequently, either intentionally through misuse or accidental as EG has a sweet taste. EG has in itself a low toxicity, but is in vivo broken down to higher toxic organic acids which are responsible for extensive cellular damage in various tissues caused principally by the metabolites glycolic acid and oxalic acid. The most conclusive analytical method of diagnosing EG poisoning is determination of EG concentration. However, victims are sometimes admitted at a late stage to hospitals or died during emergency treatment like a gastric lavage or found rotten dead, when blood EG concentrations are low or not detected. Therefore, in this study, the identification of EG was not only performed by gas chromatograpyc-mass spectrometry (GC-MS) following derivatization but also further toxicological analyses of metabolites, glycolic acid (GA) and oxalic acid (OA), were performed by ion chromatography in various biological specimens. A ranges of blood concentrations (3 cases) was $10\sim2,400\;{\mu}g/mL$ for EG, $224\sim1,164\;{\mu}g/mL$ for GA and ND $\sim40\;{\mu}g/mL$ for OA, respectively, In other biological specimens (liver, kidney, bile and pleural fluid), a range of concentrations (3 cases) was ND $\sim55,000\;{\mu}g/mL$ for EG, ND $\sim1,124\;{\mu}g/mL$ for GA and ND $\sim60\;{\mu}g/mL$ for OA, respectively. Liver and kidney tissues were recommended specimens including blood because OA, a final metabolite of EG, was identified large amounts in these despite no detectable EG caused by some therapy.

THE EFFECT OF LACTIC ACID CONCENTRATION AND pH OF LACTIC ACID BUFFER SOLUTIONS ON ENAMEL REMINERALIZATION (완충 용액의 유산 농도와 pH가 법랑질의 재광화에 미치는 영향)

  • Kwon, Jung-Won;Suh, Duk-Gyu;Song, Yun-Jung;Lee, Yun;Lee, Chan-Young
    • Restorative Dentistry and Endodontics
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    • v.33 no.6
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    • pp.507-517
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    • 2008
  • There are considerable in vitro and in vivo evidences for remineralization and demineralization occurring simultaneously in incipient enamel caries. In order to "heal" the incipient dental caries, many experiments have been carried out to determine the optimal conditions for remineralization. It was shown that remineralization is affected by different pH, lactic acid concentrations, chemical composition of the enamel, fluoride concentrations, etc. Eighty specimens from sound permanent teeth without demineralization or cracks, 0.15 mm in thickness, were immersed in lactic acid buffered demineralization solutions for 3 days. Dental caries with a surface zone and subsurface lesion were artificially produced. Groups of 10 specimens were immersed for 10 or 12 days in lactic acid buffered remineralization solutions consisting of pH 4.3 or pH 6.0, and 100, 50, 25, or 10 mM lactic acid. After demineralization and remineralization, images were taken by polarizing microscopy (${\times}100$) and micro-computed tomography. The results were obtained by observing images of the specimens and the density of the caries lesions was determined. 1.As the lactic acid concentration of the remineralization solutions with pH 4.3 was higher, the surface zone of the carious enamel increased and an isotropic zone of the subsurface lesion was found. However, the total decalcification depth increased at the same time. 2.In the remineralization solutions with pH 6.0, only the surface zone increased slightly but there was no significant change in the total decalcification depth and subsurface zone. In the lactic acid buffer solutions with the lower pH and higher lactic acid concentration, there were dynamic changes at the deep area of the dental carious lesion.

Inhibitory Effects of Chios Mastic Gum on Gastric Acid Secretion by Histamine-Related Pathway in a Rat Model and Primary Parietal Cells (위염 동물모델과 위 벽세포에서 히스타민 경로를 통한 매스틱검(Chios Mastic Gum)의 위산 분비 억제효과 및 기전 연구)

  • Nam, Da-Eun;Kim, Ok Kyung;Shim, Tae Jin;Lee, Jum Kyun;Hwang, Kwon-Tack
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.10
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    • pp.1500-1509
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    • 2014
  • The object of this study was to investigate the inhibitory effects of chios mastic gum (MG) on gastric acid secretion in an ethanol-induced SD rat model and primary parietal cells. Rats were randomly divided into four groups: Vehicle (normal group), Control (treated with ethanol), MG50 (treated with ethanol and mastic gum at 50 mg/kg b.w), MG100 (treated with ethanol and mastic gum at 100 mg/kg b.w). Groups treated with both MG50 and MG100 showed attenuation of gastric mucosal injury, sub-epithelial loss, hemorrhaging, and gastric juice secretion. We also examined the acidity of gastric juice during gastric injury. Oral administration of both MG50 and MG100 significantly decreased acidity of gastric juice by % and %, respectively. To examine the stimulatory factors related to gastric acid secretion, mRNA expression levels of H2r, M3r, CCK2r, and $H^+/K^+$ ATPase were measured by real-time PCR. Compared with a vehicle group, mRNA expression levels of H2r, CCK2r, and $H^+/K^+$ ATPase clearly increased in the control group. However, levels of H2r, CCK2r, and $H^+/K^+$ ATPase slightly but significantly decreased in MG-treated groups compared with control. Blood level of histamine significantly decreased in MG-treated groups, which indicates the involvement of MG on in histamine-related acid secretion. To identify the mode of action of MG in regulating histamine-related pathways, intracellular level of cAMP and mRNA levels of H2r, M3r, CCK2r, and $H^+/K^+$ ATPase were measured in primary parietal cells. While mRNA levels of M3r and CCK2r remained unchanged, levels of H2r and $H^+/K^+$ ATPase significantly decreased upon MG treatment. Subsequently, intracellular levels of cAMP decreased. These results suggest that mastic gum has the ability to inhibit gastric acid secretion by regulating a histamine-related pathway.