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The Signal Transduciton of Ginsenosides, Active Ingredients of Panax ginseng, in Xenopus oocyte: A Model System for Ginseng Study

  • Nah Seung-Yeol;Lee Sang-Mok
    • Proceedings of the Ginseng society Conference
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    • 2002.10a
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    • pp.66-83
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    • 2002
  • Recently, we have provided evidence that ginsenosides, the active components of Panax ginseng, utilize pertussis toxin (PTX)-insensitive $G{\alpha}_{q/11}-phospholipase\;C-{\beta}3(PLC-{\beta}3)$ signal transduction pathway for the enhancement of $Ca^{2+}-activated\;Cl^{-}$ current in the Xenopus oocyte (British J. Pharmacol. 132, 641-647, 2001; JBC 276, 48797-48802, 2001). Other investigators have shown that stimulation of receptors linked to $G{\alpha}-PLC$ pathway inhibits the activity of G proteincoupled inwardly rectifying $K^+$ (GIRK) channel. In the present study, we sought to determine whether ginsenosides influenced the activity of GIRK 1 and GIRK 4 (GIRK 1/4) channels expressed in the Xenopus oocyte, and if so, the underlying signal transduction mechanism. In oocyte injected with GIRK 1/4 channel cRNAs, bath-applied ginsenosides inhibited high potassium (HK) solution-elicited GIRK current $(EC_{50}:4.9{\pm}4.3\;{\mu}g/ml).$ Pretreatment of the oocyte with PTX reduced the HK solution-elicited GIRK current by $49\%,$ but it did not alter the inhibitory ginsenoside effect on GIRK current. Prior intraoocyte injection of cRNA(s) coding $G{\alpha}_q,\;G{\alpha}_{11}\;or\;G{\alpha}_q/G{\alpha}_{11},\;but\;not\;G{\alpha}_{i2}\;or\;G{\alpha}_{oA}$ attenuated the inhibitory ginsenoside effect. Injection of cRNAs coding $G{\beta}_{1{\gamma}2}$ also attenuated the ginsenoside effect. Similarly, injection of the cRNAs coding regulators of G protein signaling 1, 2 and 4 (RGS1, RGS2 and RGS4), which interact with $G{\alpha}_i\;and/or\;G{\alpha}_{q/11}$ and stimulates the hydrolysis of GTP to GDP in active GTP-bound $G{\alpha}$ subunit, resulted in a significant reduction of ginsenoside effect on GIRK current. Preincubation of GIRK channel-expressing oocyte in PLC inhibitor (U73122) or protein kinase C (PKC) inhibitor (staurosporine or chelerythrine) blocked the inhibitory ginsenoside effect on GIRK current. On the other hand, intraoocyte injection of BAPTA, a free $Ca^{2+}$ chelator, had no significant effect on the ginsenoside action. Taken together, these results suggest that ginsenosides inhibit the activity of GIRK 1/4 channel expressed in the Xenopus oocyte through a PTX-insensitive and $G{\alpha}_{q/11}$-,PLC-and PKC-mediated signal transduction pathway.

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Isolation of Egg-Contaminating Bacteria and Evaluation of Bacterial Radiation Sensitivity (계란 오염 세균의 분리 및 분리 균주의 감마선 감수성 평가)

  • Kim, Dong-Ho;Yun, Hye-Jeong;Song, Hyun-Pa;Lim, Byung-Lak;Jo, Cheo-Run
    • Food Science and Preservation
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    • v.15 no.5
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    • pp.774-781
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    • 2008
  • was performed and Staphylococcus sciuri, Bacillus cereus, Escherichia coli, Proteus mirabilis, and Enterococcus faecalis were identified. No Salmonella strain, a typical contaminant of eggs, was found. The radiation sensitivities of isolated bacteria and Salmonella typhimurium, in an inoculated model system, were expressed in $D_{10}$ values. The ranges of $D_{10}$ values shown by S. typhimurium, S. sciuri, B. cereus, E. coli, P. mirabilis, and E. faecalis were 0.365-0.399 kGy, 0.418-0.471 kGy, 1.075-1.119 kGy, 0.280-0.304 kGy, 1.132-1.330 kGy, and 0.993-1.290 kGy, respectively. The growth of all six test bacteria in eggs (inoculated at $10^6-10^7\;CFU/g$) during 3 days of post-irradiation storage at ambient conditions ($25^{\circ}C$) was recorded. S. typhimurium was eliminated by irradiation at 3 kGy, and E. coli and S. sciuri were eliminated by irradiation at 5 kGy. The viable cell counts of B. cereus, P. mirabilis, and E. faecalis in eggs showed 4-6 log reductions after irradiation with 5 kGy.

Anti-oxidative and Immunomodulating Activities of Solvent Extracts from Broccoli (Brassica oleracea) Sprouts (브로콜리 새싹 용매 추출물의 항산화 및 면역조절 활성)

  • Koh, Jong-Ho;Kim, Hoon;Hwang, Jong-Hyun;Yu, Kwang-Won
    • The Korean Journal of Food And Nutrition
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    • v.32 no.1
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    • pp.1-10
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    • 2019
  • In order to examine the functionality of broccoli sprout (Brassica oleracea, BS), solvent extracts were prepared and their anti-oxidative and immunomodulating activities were compared with those of broccoli (B). EtOH extracts (E) were potently higher than hot-water extracts (HW) in the antioxidant contents and radical scavenging activity. In particular, the total polyphenolic contents in addition to ABTS and DPPH radical scavenging activity were significantly higher in EtOH extract of broccoli sprout (BS-E; 9.15 mg GAE/g, 4.52 mg AEAC/g, and 1.14 mg AEAC/g) compared with that of broccoli (B-E; 7.83 mg GAE/g, 3.63 mg AEAC/g, and 0.97 mg/AEAC/g). Whereas, total flavonoid content was significantly higher in B-E (1.60 mg QE/g) than BS-E (1.43 mg QE/g). Anti-inflammatory activity was investigated using LPS-induced cell line model at a concentration of $10{\sim}100{\mu}g/mL$, in which all solvent extracts of both broccoli sprouts and broccoli were not toxic to RAW 264.7 cell lines. In anti-inflammatory activity of broccoli sprouts, EtOH extracts also showed significantly more potent activity than hot-water extracts in all sample concentrations tested. In addition, BS-E ($100{\mu}g/mL$) inhibited nitric oxide (NO) and IL-6 production to 60.9% and 68.9% compared with the LPS inflammation group (without extracts), whereas B-E inhibited 49.6% and 54.9%. On the other hand, in immunostimulating activity by splenocytes and macrophages, hot-water extract showed significantly higher activity than EtOH extract. Especially, BS-HW stimulated the splenocyte proliferation (1.2-fold against saline group) and IFN-${\gamma}$ production (264.39 pg/mL) at $100{\mu}g/mL$, and the production of IL-6 (1.33-fold), IL-12 (1.09-fold) and TNF-${\alpha}$ (1.49-fold) from macrophages was also significantly enhanced over broccoli. In conclusion, broccoli sprouts showed more potent anti-oxidative, anti-inflammatory and immunostimulating activity than broccoli, suggesting the possibility of using broccoli sprouts as functional food materials.

Effector Memory CD8+ and CD4+ T Cell Immunity Associated with Metabolic Syndrome in Obese Children

  • Yang, Da-Hee;Lee, Hyunju;Lee, Naeun;Shin, Min Sun;Kang, Insoo;Kang, Ki-Soo
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.24 no.4
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    • pp.377-383
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    • 2021
  • Purpose: We investigated the association of effector memory (EM) CD8+ T cell and CD4+ T cell immunity with metabolic syndrome (MS). Methods: Surface and intracellular staining of peripheral blood mononuclear cells was performed. Anti-interleukin-7 receptor-alpha (IL-7Rα) and CX3CR1 antibodies were used to stain the subsets of EM CD8+ T cells, while anti-interferon-gamma (IFN-γ), interleukin-17 (IL-17), and forkhead box P3 (FOXP3) antibodies were used for CD4+ T cell subsets. Results: Of the 47 obese children, 11 were female. Children with MS had significantly higher levels of serum insulin (34.8±13.8 vs. 16.4±6.3 µU/mL, p<0.001) and homeostasis model assessment of insulin resistance (8.9±4.1 vs. 3.9±1.5, p<0.001) than children without MS. Children with MS revealed significantly higher frequencies of IL-7Rαlow CD8+ T cells (60.1±19.1% vs. 48.4±11.5%, p=0.047) and IL-7RαlowCX3CR1+ CD8+ T cells (53.8±20.1% vs. 41.5±11.9%, p=0.036) than children without MS. As the serum triglyceride levels increased, the frequency of IL-7RαlowCX3CR1+ and IL-7RαhighCX3CR1- CD8+ T cells increased and decreased, respectively (r=0.335, p=0.014 and r=-0.350, p=0.010, respectively), in 47 children. However, no CD4+ T cell subset parameters were significantly different between children with and without MS. Conclusion: In obese children with MS, the changes in immunity due to changes in EM CD8+ T cells might be related to the morbidity of obesity.

Inhibitory Effect of Purified Bee Venom(Apis mellifera L.) on Adipogenesis in Korea (국내 양봉농가에서 채취한 정제봉독(Apis mellifera L.)의 지방세포 분화 억제 효과)

  • Han, Sang Mi;Kim, Hyo Young;Woo, Soon Ok;Kim, Se Gun;Choi, Hong Min;Moon, Hyo Jung
    • Journal of Apiculture
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    • v.35 no.1
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    • pp.49-54
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    • 2020
  • Bee (Apis mellifera L.) venom is used for the treatment of various human diseases due to its known anti-inflammatory and antibacterial properties. This study investigated the effect of purified bee venom (PBV) on adipogenesis in 3T3-L1 preadipocytes. There was no cytotoxicity while cells were treated with PBV by various concentrations. In the PBV treated cells increases in fat storage were inhibited and also confirmed by oil red o staining. To understand the underlying mechanism at the molecular level were examined on the expression of the genes involved in adipogenesis by using real-time PCR. In this cell model, the mRNA level of adipogenic genes such as peroxisome-proliferator-activated receptors gamma (PPARγ) and CAAAT/enhancer binding protein alpha(C/EBPα) were decreased by PAE treatment, comparing with those of control group. Theses results suggest that PBV inhibits adipocyte differentiation in 3T3-L1 cells and can be used as an efficient natural substance to manage anti-obesity.

Anti-obesity effects of herbal extract YY312 in C57BL/6 mice fed a high-fat diet and 3T3-L1 cells (백모근(白茅根) 청피(靑皮) 오수유(吳茱萸) 복합방(複合方)(YY312)의 고지방식이로 유도된 마우스와 3T3-L1 세포에서 항비만 효과)

  • Kang, Insug;Hwang, Keun-Young;Choi, A-Young;Roh, Kughwan;Choi, Ji Hyun;Sim, Yeomoon;Park, Yoo Kyoung;Oh, Myung Sook
    • The Korea Journal of Herbology
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    • v.28 no.1
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    • pp.23-31
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    • 2013
  • Objectives : The purpose of this study was to determine the anti-obesity effect and molecular mechanism of YY312, a herbal extract composed of Imperatae Rhizoma, Citri Unshius Pericarpium Immaturus, and Evodiae Fructus, on a high-fat diet-induced animal model and on 3T3-L1 cells. Methods : C57BL/6 mice were fed for 6 weeks with a normal diet or a high-fat diet (HFD). Then they orally administered daily with 300 mg/kg YY312 for next 10 weeks. Body weight and food consumption were recorded weekly and daily, respectively. Tissue weights, serum lipid, and glucose levels were analyzed at the end of the study. Additionally, the effects of YY312 on adipocyte differentiation in 3T3-L1 cells were examined. After differentiating 3T3-L1 cells were treated with YY312, Oil-red O staining, RT-PCR, and Western blotting were performed for lipid accumulation, mRNA expression of adipogenesis gene, and AMP-activated protein kinase (AMPK) phosphorylation, respectively. Results : YY312-administered mice showed a significant reduction of body weights and abdominal adipose tissue weights. YY312 also reduced the serum levels of triglycerides and total cholesterol, compared with the HFD group. Treatment with YY312 inhibited lipid accumulation and blocked expression of adipogenic transcription factors and lipogenesis genes, including peroxisome proliferator-activated receptor ${\gamma}$, CCAT/enhancer binding protein ${\alpha}$ and fatty acid synthase. YY312 increased AMPK phosphorylation in 3T3-L1 adipocytes. Conclusions : This study showed that herbal extract YY312 has an anti-obesity effect in vitro and in vivo. Thus, YY312 could be developed as a supplement for reduction of body weight gain induced by an HFD.

Non-alcoholic fatty liver protective effects, and studies on the mechanism of action of Crataegi Fructus (산사의 NAFLD 보호 효과 및 그 작용기전에 관한 연구)

  • Kim, Min-Chul;Kong, Ryong;Han, Hyoung-Sun;Kang, Dam-hee;Lee, Seung-Jin;Lee, Cheon-Cheon;Wang, Seo;Kwon, Dong-Yeul;Kang, Ok-Hwa
    • The Korea Journal of Herbology
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    • v.33 no.6
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    • pp.61-70
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    • 2018
  • Objectives : Non-alcoholic fatty liver disease (NAFLD) is characterized by the accumulation of hepatic triglycerides (TG) that leads to inflammation and fibrosis. Crataegi Fructus ethanol extract (CE) is a korean traditional herb that used for digestive diseases. It has been investigated that CE has the effect that prevent hepatotoxicity caused by CCl4 or GaIN and regulate the inflammatory in several organs. However, a hypolipidemic effect of CF has not been reported. Methods : The purpose of this study is that examine the lipid accumulation inhibitory effect of CE on NAFLD. We checked the body and liver weight change of MCD-diet induced mice with/without administration of CE. The blood lipid levels of C57BL/6J mice were checked by biochemistry. Also we observed the liver histology of MCD-diet induced mice and investigate the molecular mechanisms in MCD-diet-induced NAFLD in C57BL/6J mice. Results : CE improved MCD-diet-induced lipid accumulation and TG and TC levels. Also, CE decreased hepatic lipogenesis such as SREBP-1, $C/EBP{\alpha}$, $PPAR{\gamma}$, ACC and FAS. Besides, we also found out that CE increased AMPK phosphorylation. These results indicated that CE has the same ability to activate AMPK and then reduce SREBP-1, and FAS expression, finally leading to inhibit hepatic lipogenesis and hepatic antioxidative ability. Conclusions : In this report, we found CE exerted a regulatory effect on lipid accumulation by decreasing lipogenesis in MCD-diet induced NAFLD model. Therefore, CE extract may be active in the prevention of fatty liver.

Immune-enhancing effects of a traditional herbal prescription, Kyung-Ok-Ko (전통적인 한방 처방 경옥고의 면역 증강 효과)

  • Roh, Seong-Soo;Lee, Wonhwa;Kim, Kyung-Min;Na, MinKyun;Bae, Jong-Sup
    • The Korea Journal of Herbology
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    • v.34 no.2
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    • pp.41-47
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    • 2019
  • Objectives : A traditional herbal prescription, Kyung-Ok-Ko (KOK), has long been used in oriental medicine as an invigorant for age-related diseases, such as amnesia and stroke. However, the beneficial value of KOK for immune responses is largely unknown. Based on the above mentioned effects of KOK, other previous reports, and its use in traditional medicine, we hypothesized that KOK displays beneficial effects against methotrexate (MTX)-induced immune suppression. Methods : We investigated the effects of KOK (0.6 g/kg/day, oral (p.o.)) on deteriorated immunity caused by MTX (2 mg/kg/day, p.o.) in an immune suppression mouse model. MTX was fed to mice once a day for 7 days. After the immune responses of the mice deteriorated by MTX treatment, KOK in water was fed to the mice once a day for 14 days. We then measured the expression levels of various cytokines, such as T helper cell (Th1, Th2) cytokines, and the number of immune cells, such as spleen T cells, B cells, and macrophages. Results : The data showed that MTX decreased Th1 profiles (interferon $(IFN)-{\gamma}$, interleukin (IL)-2, IL-12) and the number of immune cells, and increased Th2 profiles (IL-4, IL-5, IL-13), which were normalized significantly by post-administration of KOK. However, there was no significant difference in body-weight gain between MTX- and KOK-treated mice. Conclusion : These results indicate that KOK has immune-enhancing functions and reduces immunotoxicity of MTX, suggesting that supplementation with KOK will improve immune responses clinically and be useful for the prevention of immune-related diseases.

Anti-Obesity Effect of Schizandrae Fructus Water Extract through Regulation of AMPK/Sirt1/PGC-1α signaling pathway (AMPK/Sirt1/PGC-1α 신호 전달 경로의 조절을 통한 오미자 추출물의 비만 개선 효과)

  • Lee, Se Hui;Park, Hae-Jin;Shin, Mi-Rae;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.37 no.2
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    • pp.1-11
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    • 2022
  • Objectives : Although the anti-obesity effect of Schizandrae Fructus water extract has been demonstrated, its underlying mechanism is still unclear. Therefore, we aimed to evaluate the anti-obesity effect of Schizandrae Fructus water extract through the p-AMP-activated protein kinase (p-AMPK), sirtuin1 (Sirt1), and peroxisome proliferator-activated receptor-gamma coactivator 1α (PGC-1α) signaling in 60% high-fat diet (HFD)-induced obese mouse model. Methods : Male C57BL/6 mice were divided into four groups. The Normal group was fed a normal diet and the obese groups were fed 60% HFD. Except for the Control group, the GG group was supplemented with 0.5% Garcinia gummigutta and the SCW group was supplemented with 0.5% Schizandrae Fructus water extract. After 6 weeks, obesity-related biomarkers in serum were measured and the expressions of protein for lipid-related factors in liver tissue were analyzed by western blot. Results : Treatment with SCW significantly down-regulated body weight compared to the Control group. SCW down-regulated levels of triglyceride and total cholesterol in serum and significantly increased p-AMPK, Sirt1, and PGC-1α in liver tissue. In addition, the expressions of fatty acid oxidation-related proteins such as peroxisome proliferator-activated receptor α (PPARα), carnitine palmitoyltransferase 1A (CPT-1A), uncoupling protein 1 (UCP1), and uncoupling protein 3 (UCP3) were significantly up-regulated. However, fatty acid synthesis-related proteins including sterol regulatory element-binding protein-1 (SREBP-1), phospho-Acetyl-CoA Carboxylase (p-ACC), and fatty acid synthase (FAS) were significantly down-regulated. Conclusions : Taken together, SCW treatment showed anti-obesity effect by regulating both fatty acid oxidation-related and fatty acid synthesis-related proteins through AMPK/Sirt1/PGC-1α signaling in 60% HFD-induced obese mice.

Cardioprotective Effect of Calcium Preconditioning and Its Relation to Protein Kinase C in Isolated Perfused Rabbit Heart (적출관류 토끼 심장에서 칼슘 전처치에 의한 심근보호 효과와 Protein Kinase C와의 관계)

  • 김용한;손동섭;조대윤;양기민;김호덕
    • Journal of Chest Surgery
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    • v.32 no.7
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    • pp.603-612
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    • 1999
  • Background : It has been documented that brief repetitive periods of ischemia and reperfusion (ischemic preconditioning, IP) enhances the recovery of post-ischemic contractile function and reduces infarct size after a longer period of ischemia. Many mechanisms have been proposed to explain this process. Recent studies have suggested that transient increase in the intracellular calcium may have triggered the activation of protein kinase C(PKC); however, there are still many controversies. Accordingly, the author performed the present study to test the hypothesis that preconditioning with high concentration of calcium before sustained subsequent ischemia(calcium preconditioning) mimics IP by PKC activation. Material and Method : The isolated hearts from the New Zealand White rabbits(1.5∼2.0 kg body weight) Method: The isolated hearts from the New Zealand White rabbits(1.5∼2.0 kg body weight) were perfused with Tyrode solution by Langendorff technique. After stabilization of baseline hemodynamics, the hearts were subjected to 45-minute global ischemia followed by a 120-minute reperfusion with IP(IP group, n=13) or without IP(ischemic control, n=10). IP was induced by single episode of 5-minute global ischemia and 10-minute reperfusion. In the Ca2+ preconditioned group, perfusate containing 10(n=10) or 20 mM(n=11) CaCl2 was perfused for 10 minutes after 5-minute ischemia followed by a 45-minute global ischemia and a 120-minute reperfusion. Baseline PKC was measured after 50-minute perfusion without any treatment(n=5). Left ventricular function including developed pressure(LVDP), dP/dt, heart rate, left ventricular end-diastolic pressure(LVEDP) and coronary flow(CF) was measured. Myo car ial cytosolic and membrane PKC activities were measured by 32P-${\gamma}$-ATP incorporation into PKC-specific pepetide. The infarct size was determined using the TTC (tetrazolium salt) staining and planimetry. Data were analyzed using one-way analysis of variance(ANOVA) variance(ANOVA) and Tukey's post-hoc test. Result: IP increased the functional recovery including LVDP, dP/dt and CF(p<0.05) and lowered the ascending range of LVEDP(p<0.05); it also reduced the infarct size from 38% to 20%(p<0.05). In both of the Ca2+ preconditioned group, functional recovery was not significantly different in comparison with the ischemic control, however, the infarct size was reduced to 19∼23%(p<0.05). In comparison with the baseline(7.31 0.31 nmol/g tissue), the activities of the cytosolic PKC tended to decrease in both the IP and Ca2+ preconditioned groups, particularly in the 10 mM Ca2+ preconditioned group(4.19 0.39 nmol/g tissue, p<0.01); the activity of membrane PKC was significantly increased in both IP and 10 mM Ca2+ preconditioned group (p<0.05; 1.84 0.21, 4.00 0.14, and 4.02 0.70 nmol/g tissue in the baseline, IP, and 10 mM Ca2+ preconditioned group, respectively). However, the activity of both PKC fractions were not significantly different between the baseline and the ischemic control. Conclusion: These results indicate that in isolated Langendorff-perfused rabbit heart model, calcium preconditioning with high concentration of calcium does not improve post-ischemic functional recovery. However, it does have an effect of limiting(reducing) the infart size by ischemic preconditioning, and this cardioprotective effect, at least in part, may have resulted from the activation of PKC by calcium which acts as a messenger(or trigger) to activate membrane PKC.

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