This study aimed to develop the in vitro method using domestic commercial diets to estimate nutrient digestibility in dogs. The existing in vitro method were tested and compared with literature data to develop new in vitro method. The development of in vitro method progressed as follows: modification of pepsin solution to an activated form and supplementation with 1% lipase. All the in vitro method progressed to 4 hours of stomach simulation and 2 hours of small intestine simulation. In vivo digestibility was measured using the same diets as beagle dogs. The supplementation of lipase methods showed significantly improved (p < 0.05) DM, OM, and EE than the existing and modified pepsin solution methods. The correlation between in vitro and in vivo data in DM, OM, and EE digestibility was high (r2 = 0.889, 0.907, and 0.721, respectively), and the correlation between in vitro and in vivo data in CP and GE digestibility was medium (r2 = 0.681 and 0.536, respectively). The current in vitro method is similar to in vivo digestibility and helps potentially predict digestibility for dogs. In conclusion, this developed in vitro method suggests that it can help estimate the nutrient digestibility of dogs' diets without in vivo experiments.

본 연구는 과체중과 정상체중의 비글견 사이에서 미네랄 소화율의 차이를 평가하기 위해 수행되었다. 중성화 된 건강한 비글견 11마리(47.7개월령 ±0.14)의 Body condition score (BCS)를 기준으로 Normal BCS 그룹(BCS≤5, n=5)과 High BCS 그룹(BCS ≥6, n=6)의 두 그룹에 배치하였다. 시험사료는 반려견의 영양소 요구량을 충족하도록 제조하여 개체 별 에너지요구량에 맞춰 일일 2회에 나누어 14일 동안 급여하였다. 미네랄의 외관상 전장 소화율은 0.5% 산화크롬(Cr2O3)을 이용한 지시제법을 이용하여 평가되었다. 사료와 분변 내 Macro 미네랄(K, Mg, P, Na, Ca)과 Micro 미네랄(Se, Fe, Zn, Cu, Mn)의 함량은 유도결합플라즈마 분광광도계를 이용하여 분석하였다. 체중과 BCS는 Normal BCS 그룹 보다 High BCS 그룹이 유의하게 높았으며(p<0.01), 시험기간 동안 두 그룹 내 각각의 체중과 BCS는 통계적으로 유의한 변화는 관찰되지 않았다(p>0.05). 미네랄 소화율을 평가한 결과, Macro 미네랄 중에서는 마그네슘, 인, 칼슘의 소화율이 High BCS 그룹에서 높은 경향으로 관찰되었으며(p<0.1), Micro 미네랄 중에서는 High BCS 그룹에서 망간의 소화율이 유의하게 높았고(p<0.05), 셀레늄과 아연의 소화율은 높은 경향(p<0.1)을 나타내었다. 또한, 통계적인 유의성과 관계없이 분석한 모든 미네랄 지표에서 High BCS 그룹이 Normal BCS 그룹보다 높은 소화율의 결과를 보여 주었다. 본 연구의 결과는 비글견에서 과체중 또는 비만이 미네랄 소화율의 변화를 유도할 잠재적 가능성을 보여주었다.

본 연구는 위장 단계의 소화과정에 관여하는 Gastric lipase (GL)를 반려견을 위한 정적 체외 소화모델(Static in vitro digestion model)에 적용을 검토하기 위하여 실시되었다. GL의 첨가가 체외 소화과정 동안 건물(Dry matter; DM), 조단백질(Crude protein; CP) 그리고 조지방(Ether extracts; EE) 소화율에 미치는 영향을 평가하였다. GL은 위장 소화단계에서 첨가되었다. 위장(39℃, 2 hr.)과 소장(39℃, 4 hr.) 소화 후에 비소화 분획을 분리하였다. 그리고 실험사료와 분리된 비소화 분획에서 DM, CP 그리고 EE 수준을 측정하고 각각의 소화율을 계산하였다. 위장과 소장 소화단계에서 측정된 DM, CP 그리고 EE 소화율은 Control과 GL 그룹 사이에서 통계적으로 유의한 차이는 관찰되지 않았다(p>0.05). 결과적으로 우리의 체외 소화모델에서 GL의 첨가는 DM, CP 그리고 EE의 소화율에는 영향을 미치지 않는 것으로 나타났다. 따라서 이와 같은 결과는 정적 체외 소화모델을 이용한 소화율의 평가에 있어서 GL의 역할은 다소 제한적일 수 있다는 것을 시사한다.

바다에 기름오염 사고가 발생하면 여러 가지 방제 방법 중 물리적 회수 방법을 우선적으로 사용하고 유처리제는 최후 수단으로 고려하는 경향이 있다. 유처리제는 수중으로 기름이 신속히 분산되도록 하여 해수면으로부터 제거하는 방법이다. 해수면으로부터 신속히 기름을 제거하는데 대한 유처리제의 효용성은 널리 증명되어 왔으나 아직도 대부분의 국가들은 해양환경에 미치는 독성을 우려하여 적극적인 사용을 하지 않고 있는 실정이다. 보고된 자료에 의하면 유처리제 사용 후 수중생물에 대한 독성은 발견되지 않았으며, 유처리제와 혼합된 기름이 기 름 그 자체보다 독성이 더 크게 나타나지 않았다. 멕시코만 기름유출 사고 시 미국 정부와 BP사는 최대한 해안에 기름이 도달하지 않는데 중 점을 두고 해수면뿐만 아니라 수중의 기름에 대해서도 유처리제를 사용하였다.

Amino acid transporters are essential for the growth and proliferation in all living cells. Among the amino acid transporters, the system L amino acid transporters are the major nutrient transport system responsible for the Na+-independent transport of neutral amino acids including several essential amino acids. The L-type amino acid transporter 1 (LAT1) is over-expressed to support cell growth in malignant tumors. The double stranded RNA-mediated RNA interference (RNAi) analysis can be in a wide variety of eukaryotes to induce the sequence-specific inhibition of gene expression. In this study, we examined the effect of LAT1 short interfering RNA (siRNA) on cell growth using siRNA of LAT1 in the KB human oral squamous cell carcinoma. In the RT-PCR analysis and western blot analysis, the siRNA of LAT1 inhibited expressions of LAT1 mRNA and protein. The uptake of [14C]L-leucine was inhibited by siRNA of LAT1. In the MTT assay, the siRNA of LAT1 inhibited the growth of the KB cells in the time-dependent manner, indicating that the growth inhibition of KB cell by the siRNA of LAT1 is induced by the blocking of neutral amino acid transport mediated by LAT1. These results suggest that the transport of neutral amino acids including several essential amino acids into the KB human oral squamous cell carcinoma is mediated mainly by LAT1. Further, the LAT1 would be a new target for the inhibition of cancer cell growth.

본 연구는 에탄올과 염산으로 유도된 위염모델에 대해 익모초의 물 추출물이 위염예방효과를 나타내는지 알아보기 위해 진행하였고, 안전성 평가를 위해 유전독성평가인 소핵시험을 수행하였다. 익모초 물 추출물은 음성대조군 대비 위에서 분비되는 PGE2의 농도를 증가시켰을 뿐만 아니라 염산과 에탄올 투여에 의한 점막 표피세포 및 선상피세포의 손상과 울혈을 충분히 예방하는 것을 확인하였다. 또한 익모초 물 추출물에 대한 소핵시험을 실시한 결과, 익모초 물 추출물은 소핵을 유발하지 않는 것으로 나타났다. 결과를 종합하였을 때 익모초 물 추출물은 자체적으로 위염 예방효과를 나타냈지만 정확히 어떤 기전을 통해 예방하는지 추가적인 실험이 필요하다고 사료되며, 익모초에 포함된 여러 단일 성분을 이용해 위염 예방평가를 실시하여, 위염에 대한 익모초의 효능과 효율을 높이는 연구가 필요하다고 생각된다.

Background: Taraxacum platycarpum has been used in traditional medicine in Korea to treat intoxication and edema and as a diuretic. According to previous reports, it has anti-cancer, anti-gastritis, and anti-inflammation effects. However, the improvement effect of T. platycarpum on rheumatoid arthritis has not been investigated. The anti-oxidative and anti-inflammation effects of the aerial parts of T. platycarpum are different from those of its subterranean parts. Thus, we evaluated the effect of the water extracts of Taraxaci radix (WTR) on type II collagen-induced rheumatoid arthritis (CIA) in animal models. Methods and Results: Rheumatoid arthritis was induced by type II collagen. WTR (100㎎/㎏ and 500㎎/㎏) was administered to the animal models. Methotrexate was used as the positive control. The levels of interleukin-6, TNF-alpha, and type II collagen IgG in the animals were measured by using enzyme-linked immunosorbent assay. Treatment with 500㎎/㎏ WTR decreased the serum levels of interleukin-6, TNF-alpha, and collagen IgG in the CIA models. Moreover, treatment with WTR diminished the arthritisinduced swelling of the hind legs and monocyte infiltration in the bloodvessels of the animal models. Conclusions: These results indicate that WTR has the potential to improve rheumatoid arthritis by reducing the levels of inflammatory cytokines such as interleukin-6 and TNF-alpha. However, further experiments are required to elucidate the influence of WTR on signal transduction in vitro and in vivo.

Lung cancer, the most common malignant disease worldwide, is the predominant cause of cancer deaths, particularly amongst men. Therefore, various researchers have focused on the growth inhibitory effects of medicinal plants used in traditional Korean medicine. This study aimed to investigate the growth inhibitory effects of ethanol extracts of Rubiae radix, Inulae flos, Nelumbinis receptaculum, Astilbe radix, and Lagerstroemia flos on NCI-H1229 cells. Method and Results: The viability of NCI-H1229 cells was evaluated in vitro using an MTS assay. Treatment with the ethanol extracts of the selected medicinal plants at 500 ㎍/㎖ reduced NCI-H1229 cell viability and increased apoptotic cell death and caspase-3 activation. In addition, treatment with ethanol extracts of Inulae flos and Astilbe radix increases DNA fragmentation, as measured by the TUNEL assay. Conclusions: These results indicated that ethanol extracts of Rubiae radix, Inulae flos, Nelumbinis receptaculum, Astilbe radix, and Lagerstroemia flos exhibited growth inhibitory effects, inducing apoptotic cell death, DNA fragmentation and caspase-3 activation in NCI-H1229 cells. Therefore, these medicinal plant extracts may be used in the development of natural medicines to inhibit the growth of lung cancers. However, further study is needed to determine the active ingredients of the ethanol extracts from medicinal plants that are reposible for the inhibitory effect on lung cancer cell grwoth.

Background: Cassia tora L., an annual or perennial plant of the Fabaceae family, is traditional medicine with various biological activities, including anti-constipation and, anti-inflammation. Chemical compounds such as anthraquinone glycoside and naphthalene derivatives have been isolated from this plant. Cassia tora L. is a common contaminant of agricultural commodities, but is toxic to cattle and poultry. Methods and Results: To investigate the potential toxicity, Cassia tora L. aqueous extract (CO) was administered orally to rats for 26 weeks at 0 (control), 300, 1,500 and 3,000㎎/㎏/day (n = 10 for male rats for each dose). The positive control comprised animals orally administered anthraquinone 100㎎/㎏/day. There was no treatment-related mortality. An increase in the kidney weight was observed at 3,000㎎/㎏/day of CO and anthraquinone 100㎎/㎏/day. Macrophage infiltration in the colon was observed at CO 1,500 and 3,000㎎/㎏/day and anthraquinone 100㎎/㎏/day, but there were no significant toxicological changes in the incidence and severity of the finding. Conclusions: The oral no-observed-adverse-effect level (NOAEL) of CO was 3,000㎎/㎏/day in male rats and no target organs were identified. In addition, 300㎎/㎏ was found to be the no-observed-effect level (NOEL) for systemic toxicity under the conditions of the study.

Background: Astilbe chinensis (Maxim.) Franch. Et Savat. is a plant belonging to Saxifragaceae family and contains various active ingredients including astilbin and bergenin. It has been used as a traditional Korean medicine to improve fever, pain, and cough. Recently, a number of Korean medical resources have been studied for cancer and inflammation treatment, but A. chinensis (Maxim.) Franch. Et Savat. has not yet been investigated. Consequently, this study investigated the inhibitory effect of ethanol extracts from A. chinensis (Maxim.) Franch. Et Savat. (ARE) on oxidative stress and colorectal cancer using RAW264.7 and the human colorectal cancer cell line HCT-116. Methods and Results: In total, 500 ㎍/㎖ ARE reduced cell viability by 38.96 ± 1.32%, and increased caspase-3 activity by 133.08 ± 3.41% in HCT-116 cells. Moreover, TUNEL signaling and the early apoptosis ratio (34.56 ± 1.67%) increased by 500 ㎍/㎖ ARE treatment. H2O2-induced oxidative stress and cell death were diminished by 500 ㎍/㎖ ARE treatment through decreasing ROS (reactive oxygen species). Conclusions: The inhibitory effects of ARE against human colorectal cancer cells is mediated by apoptosis and caspase-3 activation, and H2O2-induced ROS generation and cell death are decreased by ARE treatment in RAW264.7 cells. However, further study is required to explore how ARE treatment is involved in the signaling pathway to decrease ROS.

Erectile dysfunction (ED), also known as impotence, is the inability to attain and sustain an erection firm enough to have sexual intercourse. Frequent ED may be a symptom of health problems including heart disease, obesity, alcoholism, stress, smoking, and depression, that need treatment. This study aimed to effect of complex extract (CPL) including Cornus officinalis on sexual function factor in the erectile dysfunction rat model. The erectile dysfuction rat model was induced by cimetidine (500 ㎎/㎏ in 5% ethanol, oral injection 2 weeks). Rats were oral administered with different concentration of CPL in rat erectile dysfunction model. As a results, sexual function factors (NO, cGMP) significantly improved in CPL treated groups (CPL-300, 600, 900 ㎎/㎏) compared to CON group. Serum testosterone was increased in a dose-dependent manner after CPL treatment. Furthermore, administrations of CPL restored lumen areas of the prostate in the erectile dysfunction rat model. These results indicated that CPL alleviated erectile dysfunction by increasing sexual function factor and testosterone in rat model. CPL could be used to natural treatement for erectile dysfunction. However, further study is required to identify active ingredient and its mechanism of erectile dysfunction.

Rubiae radix is root of Runia akane Nakai, it has been used to hemostasis and blood stasis in Korean and China. This study investigated that anti-oxidant and anti-colorectal cancer effect of ERA (ethanol extract of Rubiae radix) and WRA (water extract of Rubiae radix) using RAW 264.7 (murine macrophage from blood) and HCT-116 cells (human colorectal cancer cell line). ERA contained polyphenol (45.77 ± 2.03 ㎎/g) and flavonoid (22.82 ± 1.33 ㎎/g). 500 μM H2O2-induced ROS generation was diminished by 500 ㎍/㎖ ERA treatment in RAW 264.7 cells, but not WRA (125, 250, and 500 ㎍/㎖). Moreover, caspase-3 activity and DNA fragmentation increased by 500 ㎍/㎖ ERA treatment during apoptotic cell death in HCT-116. Results demonstrated that anti-cancer effect of ERA against human colorectal cancer cells is mediated apoptotic cell death and DNA fragmentation through caspase-3 activation. However, further study is required to what active ingredient of ERA are important for anti-oxidant and anti-colorectal cancer effect in vivo.

Doxorubicin is a anti-cancer drugs that interferes with the growth and spread of cancer cells in human body. Doxorubicin is used to treat different types of cancers that affect the ovary, thyoid and lungs, but induced side effect such as nephrotoxicity and cardiotoxicity. Thus, we investigated that the effect of iridin on doxorubicin-induced necrosis in HK-2 cells, a human proximal tubule cell. To confirm effect of iridin on doxorubicin-induced necrosis, HK-2 cells are treated with 10 μM doxorubicin and 80 μM iridin. 80 μM iridin reduced 10 μM doxorubicin-induced necrosis, the mitochondrial over activation and caspase-3 activation. However, iridin reduces anti-cancer effect of doxorubicin such as PARP1 and caspase-3 activation, checkpoint proteins (CDK4 and CDK6) in NCI-H1129 cells (Human non-small cell lung cancer cell). In HCT-116 cells (Human colorectan cancer cell), iridin do not increased protein expression of CDK4 and CDK6 decreased by doxorubicin. Results indicate that treatment of iridin was diminished doxorubicin-induced necrosis in HK-2 cells. However, iridin was decreased anti-cancer effect of doxorubicin on NCI-H1229, but not HCT-116. Thus, further experiment are required to iridin treatment on various cancer cells and animal models because effect of iridin different cell type.

Background: Inula japonica Thunb. is a plant belonging to the family compositae. Inulae flos (flower of I. britannica var. chinensis Regal.) is the dried flower of I. japonica Thunb. and contains various flavonoids (patulitrin, nepitrin and kaempferol), which have been utilized in traditional oriental medicine to treat nausea, phlegm, and coughs. However, ethanol extract of I. britannica (IJE) has not been previously studied for its use in cancer treatment, and its effects on oxidative stress, or inflammation. Thus, the present study investigated the anti-oxidant, anti-inflammatory, and anti-colorectal cancer effects of IJE using RAW264.7 and HCT- 116 cells, which are human colorectal cancer cell line. Methods and Results: IJE contained flavonoids (80.95 ± 5.3 ㎎/g) and polyphenols (310.53 ± 10.6 ㎎/g). Moreover, it reduced lipopolysaccharide (LPS)-induced nitric oxide (NO) production and H2O2-induced oxidative stress by decreasing reactive oxygen species (ROS) levels. Additionally, the 500 ㎍/㎖ IJE treatment increased caspase-3 activity and apoptotic cell death in HCT-116 cells. Conclusions: These results demonstrate that the anti-cancer effect of IJE against human colorectal cancer cells involves caspase-3 activation and apoptotic cell death. IJE also inhibited LPS-induced NO production, and H2O2-induced oxidative stress in RAW264.7 cells. However, further studies are required to explore how IJE treatment regulates signal transduction in NO and ROS production.

Background: Cynaroside is a flavone, a flavonoid-like compound, known by different names (luteoloside and cinaroside). It is commonly found in Lonicera japonica Thunb., Chrysanthemum moriflium, and Angelica keiskei. The process of cell death has been classified as necrosis and apoptosis. Necrosis refers to unregulated cell death induced by a chemotherapeutic agent. Doxorubicin is an anthracycline anti-cancer drug used to treat acute leukemia, cancer, and lymphoma. However, it induces nephrotoxicity including tubular damage. Therefore, we investigated the protective effect of cynaroside against doxorubicin-induced necrosis in HK-2 cells. Methods and Results: To confirm the beneficial effect of cynaroside on doxorubicin-induced necrosis, HK-2 cells, a human proximal tubule epithelial cell line were treated with 10 μM doxorubicin and 80 μM cynaroside. Doxorubicin treatment resulted in increased DNA fragmentation, caspase-3 activity and mitochondria hyperactivation during cell necrosis. However, pretreatment with 80 μM cynaroside attenuated DNA fragmentation, caspase-3 activity and mitochondria hyperactivation induced by 10 μM doxorubicin in HK-2 cells. Conclusions: These results indicated that pretreatment with cynaroside ameliorated doxorubicin-induced necrosis in HK-2 cells. Therefore, cynaroside be used as a therapeutic agent for improving doxorubicin-induced nephrotoxicity. However, further studies are required to evaluated the toxicity of cynaroside treatment in animals and to determine its protective effect against doxorubicininduced nephrotoxicity in an animal model.

Background: Cisplatin is one of the most extensively used chemotherapeutic agents for the treatment of cancer, including bladder, and ovarian cancers. However, it has been shown to induce nephrotoxicity, despite being an outstanding anti-cancer drug. In this study, we investigated the protective effect of dopaol β-D-glucoside (dopaol) on cisplatin-induced nephrotoxicity. Methods and Results: To confirm the protective effect of dopaol on cisplatin-induced nephrotoxicity, HK-2 cells were treated with 20 μM cisplatin and 80 μM dopaol. Cisplatin increased apoptosis, caspase-3 activity and mitochondrial dysfunction; however pretreatment with 80 μM dopaol successfully attenuated apoptosis, caspase-3 activity and mitochondrial dysfunction. To evaluate the protective effect dopaol on cisplatin-induced nephrotoxicity in vivo, we used an animal model (balb/c mice, 20 ㎎/㎏, i.p. once/day for 3 day). The results were similar to those obtained using HK-2 cells; renal tubular damage and neutrophilia induced by cisplatin reduced following dopaol injection (10 ㎎/㎏, i.p. once/day for 3 day). Conclusions: These results indicate that dopaol treatment reduced cisplatin-induced nephrotoxicity in vitro and in vivo, and can be used to treat cisplatin-induced nephrotoxicity. However, further studies are required to determine the toxicity high dose dopaol and the signal pathways involved in its mechanism of action in animal models.

Background: Constipation is one of the most common functional gastrointestinal disorder. The present study examined the ability of water extract of fermented (FRC) and non-fermented (NFRC) roasted Cassia tora to improve intestinal function and reduce constipation in a rat constipation model.Methods and Results:Different concentration of FRC and NFRC were orally administered loperamide (5 ㎎/㎏; LOP) reduced the number, weight, and water content of feces, as well as intestinal transit motility. However, 24 h-(24 hour fermented roasted-Cassia tora) 300 ㎎/㎏ FRC administration increased the number, weight, and water concent of feces, compared to that seen in the LOP group, and also improve intestinal transit mitility and, the thickness of distal colon and mucous fluid.Conclusions:The results of the present study indicated that LOP-induced constipation was improved by treatment with FRC. Therefore FRC could be used to develop functional foods or natural medicine for constipation. However, further study is needed to clarify how fermentation improves the medicinal properties of roasted C. tora.

Background: Sedum takesimense Nakai has been used as folk medicine in Korea. The present study aimed to determine the biological activity of S. takesimense by investigating the anti-inflammatory effects of S. takesimense water extract (SKLC) on the lipopolysaccharide-induced inflammatory response in RAW 264.7 cells. Methods and Results: Cytotoxicity of SKLC on RAW 264.7 cells was determinded by performing MTS assay was found to have no cytotoxic effect on RAW 264.7 cells at a concentration range of 62 - 500 ㎍/㎖. Further, pretreatment of SKLC inhibited lipopolysaccharide-induced nitric oxide (NO) production in a dose-dependent manner. To determined the inhibitory mechanisms of SKLC on inflammatory mediators, we assessed the inducible nitric oxide synthase (iNOS) and cyclooxygnease-2 (COX-2) pathways. The activities of these pathways were decreased in a dose-dependent manner by SKLC. The production of tumor necrosis factor- α (TNF-α), interleukin (IL)-1β‚ and IL-6 were also reduced. Conclusions: These results suggest that the down regulation of iNOS, COX-2, TNF-α, IL-1β‚ and IL-6 expression by SKLC are mediated by the down regulation of nuclear factor-κB (NF-κB) activity, a transcription factor necessary for pro-inflammatory mediators. This might be the mechanism underlying the anti-inflammatory effects of SKLC.