• 대한한의학회지
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• 제28권4호
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• pp.114-123
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• 2007

Osteoarthritis is characterized by cartilage degradation and chondrocytes death. Chondrocyte death is induced by the apotosis through special mechanisms including the activation of caspase-3. On the basis of this background, this study was designed to examine the cartilage protective and anti-apototic effects of Aralia Cordata in in vtro and in collagenase-induced arthritis rabbit model. To conduct in vitro study, chondrocytes culturedfrom rabbit knee joint were treated by 5 ng/ml IL-1a.For in vivo experiment, collagenase-induced arthritis (CIA) rabbit model was made via intraarticular injection with 0.25 ml of collagenase solution. Aralia cordata pharmacopuncture (ACP) was administrated on bilateral Dokbi acupoint (ST35) of rabbits at a dosage of 150 ${\mu}g/kg$ once a day for 28 days after the initiation of the CIA induction. In the study by using CIA rabbit model in vivo, ACP showed the inhibition of cartilage degradation in histological analysis. Aralia cordata also showed anti-apoptotic effect both in vitro and in vivo study. In chondrocytes treated by IL-1a, Aralia cordata inhibited caspase-3 activity and enhanced the proliferation of IL-1a-induced dedifferentiated chondrocytes. ACP showed the inhibition effect on the caspase-3 expression and activity from CIA rabbit model. This study indicates that ACP inhibits the cartilage destruction and the chondrocyte apotosis through downregulation of caspase-3 activity. These data suggest that ACP has a beneficial effect on preventing articular cartilage destruction in osteoarthrtis.

• Biomolecules & Therapeutics
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• 제29권6호
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• pp.615-629
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• 2021

An active compound, triterpene saponin, astersaponin I (AKNS-2) was isolated from Aster koraiensis Nakai (AKNS) and the autophagy activation and neuroprotective effect was investigated on in vitro and in vivo Parkinson's disease (PD) models. The autophagy-regulating effect of AKNS-2 was monitored by analyzing the expression of autophagy-related protein markers in SH-SY5Y cells using Western blot and fluorescent protein quenching assays. The neuroprotection of AKNS-2 was tested by using a 1-methyl-4-phenyl-2,3-dihydropyridium ion (MPP⁺)-induced in vitro PD model in SH-SY5Y cells and an MPTP-induced in vivo PD model in mice. The compound-treated SH-SY5Y cells not only showed enhanced microtubule-associated protein 1A/1B-light chain 3-II (LC3-II) and decreased sequestosome 1 (p62) expression but also showed increased phosphorylated extracellular signal-regulated kinases (p-Erk), phosphorylated AMP-activated protein kinase (p-AMPK) and phosphorylated unc-51-like kinase (p-ULK) and decreased phosphorylated mammalian target of rapamycin (p-mTOR) expression. AKNS-2-activated autophagy could be inhibited by the Erk inhibitor U0126 and by AMPK siRNA. In the MPP⁺-induced in vitro PD model, AKNS-2 reversed the reduced cell viability and tyrosine hydroxylase (TH) levels and reduced the induced α-synuclein level. In an MPTP-induced in vivo PD model, AKNS-2 improved mice behavioral performance, and it restored dopamine synthesis and TH and α-synuclein expression in mouse brain tissues. Consistently, AKNS-2 also modulated the expressions of autophagy related markers in mouse brain tissue. Thus, AKNS-2 upregulates autophagy by activating the Erk/mTOR and AMPK/mTOR pathways. AKNS-2 exerts its neuroprotective effect through autophagy activation and may serve as a potential candidate for PD therapy.

• 한국가축번식학회지
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• 제21권2호
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• pp.177-183
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• 1997

The present study examined the effects of epidermal growth factor(EGF) and transforming growth factor-$\alpha$(TGF-$\alpha$) on in vitro maturation of porcine follicular oocytes. Basic medium used TCM-HEPES, and oocytes cultured for 42 hours in vitro. The results obtained are as follows; 1. The nuclear maturation rates of EGF-treated groups(10ng/ml, 75.9% ; 30ng/ml, 69.2% ; 50ng/ml, 67.2% ; 100ng/ml, 71.0%) on the porcine oocytes cultured in medium without pFF in vitro were significantly(P<0.01) higher than those of non-treated group(57.1%). When the oocytes were cultured in media with 10%(v/v) pFF, the nuclear maturation rates of 30ng EGF/ml(77.1%) treated group were significantly(P<0.01) higher than those of non-(59.2%) and EGF-treated groups(10ng/ml, 65.4% ; 50ng/ml, 65.5% ; 100ng/ml, 70.4%). 2. The nuclear maturation rates of 30ng TGF-$\alpha$/ml treated group(71.9%) in media without pFF in vitro were significatnly(P<0.01) higher than those of non-(57.1%) and TGF-$\alpha$ treated groups(10ng/ml, 60.4% ; 50ng/ml, 65.4% ; 100ng/ml, 60.0%). When the oocytes were cultured in media with 10%(v/v) pFF, the nuclear maturation rates of 30 and 50ng TGF-$\alpha$/ml(77.4% and 79.6%) treated groups(10ng/ml, 64.2% ; 100ng/ml, 61.6%). 3. On the effect of EGF(30ng/ml) and/or TGF-$\alpha$(30ng/ml) treated groups in medium without pFF in vitro, the nuclear maturation rates indicated 57.3, 60.4, 75.9 and 79.7% in media with no EGF & TFG-$\alpha$, TGF-$\alpha$ only, EGF only nad EGF＋TGF-$\alpha$ treated groups, respectively. The nuclear maturation rates in medium with EGF only or EGF＋TGF-$\alpha$ were significantly(P<0.01) higher than those non- and TGF-$\alpha$ treated groups. When the oocytes were cultured in media with 10%(v/v) pFF, the nuclear maturation ratesof EGF＋TGF-$\alpha$ treated group(75.9%) were significantly(P<0.01) higher than those of non-(59.2%), TGF-$\alpha$ only (64.2%) and EGF only(69.4%) treated groups.

• Natural Product Sciences
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• 제4권1호
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• pp.26-31
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• 1998

We have examined in vitro and in vivo radioprotective effect of a Chinese medicinal plants prescription, Lifukang. Micronucleus assay was employed to evaluate in vitro radioprotective effect of Lifukang. In the presence of Lifukang, the frequencies of miconuclei were greatly reduced from 7.2 to 2.9, 1.6 and 1.6% at the concentrations of Lifukang from 0 to 2, 10 and $50{\mu}g/ml$, respectively. For in vivo assay, we monitored the incidences of apoptotic cells in mouse small intestine crypts and endogeneous spleen colonies. When Lifukang was administered to mice P.O. Or I.P. at doses of 1 mg/ml in drinking water for 7 days or 0.3 mg/mouse 24 hrs prior to irradiations, respectively, the average numbers of apoptotic cells were reduced to 3.1 or 2.3, respectively, as compared to 4.4 acquired from untreated control experiments. In addition, in spleen colony assay, Lifukang increased the number of hematopoietic spleen colonies. When samples were administered after irradiation, better results were obtained. The numbers of spleen colonies were increased from 14 colonies to 18.3 or 19.6 colonies when Lifukang was given through P.O. (1 mg/ml in drinking water for 11 days) or I.P. (0.3 mg/ mouse) after irradiation, respectively.

• 한국수정란이식학회지
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• 제23권4호
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• pp.245-250
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• 2008

This study was designed to investigate the effect of essential amino acids (EAA) and/or non-essential amino acids (NEAA) on the development of parthenogenetic and somatic cell nuclear transfer (SCNT) porcine embryos in vitro. To evaluate the timing of amino acids supplementation, activated oocytes were cultured in NCSU23-PVA with EAA, NEAA or NEAA+EAA (AAs) during specific periods as below: EAA, NEAA or AAs were supplemented during Day 0 to 6 (whole culture period: ALL), Day 2 to Day 6 (post-maternal embryonic transition period: POST-MET), Day 5 to Day 6 (post-compaction period: POST-CMP), Day 0 to Day 2 (pre-maternal embryonic transition period: PRE-MET), or Day 0 to Day 4 (post-compaction period: PRE-CMP). Supplementation of NEAA decreased cleavage rates in PRE-MET and PRE-CMP and also decreased blastocyst rates in POST-CMP. On the other hand, EAA significantly enhanced blastocyst formation rate in POST-MET and no detrimental effect on embryonic development in other groups. Interestingly, NEAA and EAA had synergistic effect when they were supplemented to the medium during whole culture period. Supplementation of AAs also enhanced SCNT porcine embryo development whereas BSA-free medium without AAs could not supported blastocyst formation of SCNT embryos. In conclusion, existence of EAA and NEAA in defined culture medium variously influences the development of parthenogenetic and SCNT porcine embryos, and their positive effect are only occurred when both EAA and NEAA are supplemented to the medium during whole culture period. Additionally, AAs supplementation enhances the blastocyst formation of SCNT porcine embryos when they are cultured in the defined condition.

• 대한한의학회지
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• 제32권2호
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• pp.1-13
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• 2011

Background and Objective: Coptidis Rhizoma is a medicinal herb known for its antioxidant and anti-inflammatory effect. The purpose of this study was to examine the effects of CR on the experimental burn elicitation in vitro and in vivo. Material and Methods: In order to know the antioxidant effect on skin cell of mice after burn elicitation, superoxide dismutase (SOD) activity was measured. In vitro, the RAW 264.7 macrophage cells were treated with lipopolysaccharides for experimental inflammation. iNOS mRNA expression was observed after CR-treatment. In order to know effects on the skin regeneration in the burned mice, we counted the nitric oxide (NO) in blood. We also observed the histological structure in the epidermal basal layer and the dermal section, and we studied changes of angiogenesis in the capillaries surrounding the basal layer and dermal papilla. The changes of transcription of iNOS mRNA (inducible nitric oxide synthase mRNA) and changes of NF-${\kappa}$B (nuclear factor ${\kappa}$B) p65 positive reaction were also observed to investigate the changes of the stress in the skin. Results: The results indicated that CR has significant effects on the antioxidant effect on skin cells of mice after burn elicitation by increasing SOD activity in the in vitro test. It seemed that CR decreased the amount of NF-${\kappa}$B which induced the iNOS mRNA dose-dependently and suppress activating NO and angiogenesis. Furthermore, CR facilitated the process of skin recovery after experimental burn. Conclusion: CR can be applied for burned skin via antioxidant effect and skin regeneration.

• 생약학회지
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• 제32권1호통권124호
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• pp.22-30
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• 2001

80% extract of Gamigwaruhaebaekbaekju-Tang (GGHBT), Gagamgwaruhaebaekbaekju-Tang (GGGHBT) and Gamigwaruhaebaekwhanggum-Tang (GGHWT) remarkably showed inhibitory effects on HMG-CoA reductase, lipid peroxidation of rat liver and LDL oxidation, and DPPH free radical scavenging effect in a dose-dependent manner. Especially, GGHWT which is formulated with Trichosanthis Fructus, Pinelliae Tuber, Aurantii Immatures Fructus, Magnoliae Cortex, Allii Macrostemi Bulbus, Cinnamomi Ramulus and Scutellariae Radix on the basis of Gwaruhaebaekbaekiu-Tang listed on the traditional medicinal references showed more effective hypocholesterolemic activities in vitro bioassay than the other prescriptions.

• 한국응용과학기술학회지
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• 제2권1호
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• pp.77-81
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• 1985

In order to investigate the effect of ginseng saponins on the activity of lipoprotein lipase, it was attempted to conform the enzymatic hydrolysis of chylomicron with post-heparin induced plasma lipoprotein lipase of normal rabbit in vitro. And the activity of lipoprotein llipase was determined by the quantitative determination of liberated free fatty acids on the hydrolysis of chylomicron. As the result, it was observed that the ginseng saponins accelerated the hydrolysis of chylomicron by post-heparin plasma in vitro. And the optimum concentration of ginseng saponins for the activity of the lipoprotein lipase in the 2% bovine serum albumin was $10^{-4}%$. But ginseng saponins on the hydrolysis of chylomicron was influenced by the presence and the absence of albumin. And the optimum concentration of albumin and Na-cholate on the activity of lipoprotein lipase was each of the $10^{-6}%$ albumin and 5mM Na-cholate. From these results, it seems that ginseng saponins might stimulate the intravascular hydrolysis of chylomicron.

• The Korean Journal of Physiology and Pharmacology
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• 제6권4호
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• pp.225-233
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• 2002

Kanagawa hemolysin (KH), an exotoxin produced from Kanagawa phenomenon-positive Vibrio parahemolyticus, has been shown to possess various biological activities including hemolysis, enterotoxicity, cytotoxicity, and cardiotoxicity. The aim of this study was to investigate the effect of KH on the cardiovascular system and its mechanism, employing in vivo and in vitro experiments of the rat. Intracerebroventricular (icv) administration of 100 mHU KH produced a marked and continuous pressor effect (icv KH-pressor effect), and the icv pressor effect was not repeatable. However, intravenous (iv) injection of the same dose of KH induced a prominent depressor effect (iv KH-depressor effect). The icv KH-pressor effect was inhibited by acid-denaturation, while the iv KH-depressor effect was not. Simultaneous icv administration of the three agents (ouabain, diltiazem, or bumetanide: $10{\mu}g/kg$ each) significantly reduced the pressor effect. The icv KH-pressor effect was inhibited by treatment with iv phentolamine or chlorisondamine, but was not affected by iv candesartan. The iv KH-depressor effect was repeatable and was attenuated by treatment with iv NAME or methylene blue. In vitro experiments using isolated thoracic aorta, $10^{-6}$ M phenylephrine (PE) and 50 mM KCl produced a sustained contraction. In rings contracted with either agents, KH showed relaxant responses in a concentration- dependent fashion and the relaxation (KH-vasorelaxation) was not dependent on the existence of the endothelium. The KH-vasorelaxation in the endothelium-intact rings contracted by PE was abolished by methylene blue treatment. In summary, the present findings suggest that in the icv KH-pressor effect the cation leak-inducing action of KH is implicated, which leads to the increased central sympathetic tone, that the iv KH-depressor effect results from the vasorelaxation via NO-guanylate cyclase system, and that the KH-vasorelaxation is independent of the endothelium and the guanylate cyclase system is involved in it. In conclusion, the mechanism of KH producing the icv pressor effect may not be identical to that of KH producing the iv depressor effect.

• Asian-Australasian Journal of Animal Sciences
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• 제20권11호
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• pp.1721-1728
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• 2007

An in vitro and a feeding trial were conducted to investigate the effect of xylanase supplementation on the feeding value of growing pig diets containing high proportions of Chinese double-low rapeseed meals (DLRM). Seven diets were formulated to meet NRC (1998) nutrient requirements. Diet 1 based on corn-soybean meal was used as positive control 1, and diet 2, a practical diet which incorporated a conventional level of Chinese DLRM (60 g/kg diet), as positive control 2. Diet 3 contained a higher level of DLRM (100 g/kg diet) as the negative control. Diet 3 plus xylanase at 0.10, 0.25, 0.50 and 0.70 g/kg diet created diets 4, 5, 6 and 7, respectively. The seven diets were incubated in triplicate with the in vitro two-stage enzyme incubation method to predict responses of diets to xylanase in terms of digestibility of dry matter (DM), crude protein (CP) and neutral detergent fibre (NDF). In vitro, the negative control had the lowest CP and NDF digestibility. Both DM and CP digestibility were increased (p<0.05) owing to xylanase supplementation either at 0.50 or 0.70 g/kg diet, and NDF digestibility was improved following xylanase addition at all of the test levels. There was a high linear correlation ($r^2>90$, p<0.05) between the activity concentration of the enzyme when transformed into its logarithmic value and in vitro digestibility coefficients of DM, CP or NDF. In the feeding trial, 112 crossbred pigs were randomly assigned to seven dietary treatments with 16 replicate pens of one pig each. An obvious dose effect on growth rate was observed ($r^2=0.79$, p<0.05) within the inclusion levels of xylanase. Compared with the negative control, xylanase addition at 0.70 g/kg diet resulted in significantly increased ADG (878 g/d vs. 828 g/d, p<0.05), and a tendency towards improved growth rate (868 g/d vs. 828 g/d, p = 0.10) was also observed following the inclusion of xylanase at 0.50 g/kg diet. It would appear that the nutrient utilization of corn and Chinese DLRM diets by pigs could be enhanced by an appropriate amount of xylanase addition. The in vitro and in vivo results suggested that the in vitro incubation method is feasible for predicting responses of pigs to exogenous enzymes and identifying those preparations that possess potential for improvement of the nutritive values of feedstuffs.