• Nutritional Sciences
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• v.5 no.4
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• pp.197-202
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• 2002

We compared the radical scavenging activity of flavonoids and their antioxidant effects on erythrocyte Na leak, platelet aggregation and TBARS (thiobarbituric acid reactive substance) production, using Sprague Dawley rats. The concentrations of flavonoids needed for scavenging radicals by 50% ($SC_{50}$) in 0.1mM DPPH (2,2 Diphenyl 1-picryl hydrazyl) were: Quercetin, 7.4/$\mu$M; Catechin, 10.6$\mu$M; Morin, 22$\mu$M; Hesperidin, 400uM; and Naringin, 3.95mM. Morin completed its antioxidant activity in 2 minutes, while catechin, hesperidin and naringin had slow but long lasting antioxidant activity. Whole blood platelet aggregation, when incubated with quercetin or catechin, was significantly decreased (P<0.05) compared with the control. Sodium leak in intact erythrocytes was significantly lower when incubated with quercetin, compared with other flavonoids (P<0.05). Morin, hesperidin and naringin somewhat increased Na leak in intact erythrocytes. Sodium leak in erythrocytes treated with phenazine methosulfate (PMS) was increased overall, but was not affected by flavonoids. Intracelluar Na and K were not affected by treatment with PMS. TBARS production in platelet rich plasma (PRP) was significantly lower (P<0.05) than the control when incubated with quercetin or hesperidin. PMS treatment caused an increase in TBARS production regardless of flavonoids. In the present study antioxidant effects of flavonoids were not well correlated with their radical scavenging activities, although quercetin, which showed the strongest radical scavenging activity, had the greatest antioxidant effect.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.20 no.3
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• pp.71-81
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• 2007

Objective : In this study, herbal medicine(GJE, Gardenia jasminoides Ellis; HCT, Houttuynia cordata Thunb.; CIL, Chrysanthemum indicum Linne; PMS,Paeonia moutan Sims, P. subfruticosa Makino; APL, Agrimonia pilosa Ledebour) were screened for their inhibitory activities against Tyrosinase and PMA plus A23187-induced $TNF-{\alpha}$, IL-6, IL-8 productions in HMC-1 cells to reveal their skin -whitening and anti-allergic effect. Method : To investigate Tyrosinase inhibition we treated Mushroom Tyrosinase(Fluka, 93898) $10{\mu}{\ell}$ and 7.5mM Tyrosine (Sigma, T3754) $20{\mu}{\ell}$ with 80% ethanol medicine extracts. Then we observed 96well micro plate extinction at 490nm. In the next experiment, to investigate Anti-allergic effect we blended cultured Human Mast Cells(HMC-1) with medicine extracts. We treated the blended solution with Phorbol 12-myristate 13-acetate(PMA) and A23187, then observed $TNF-{\alpha}$, IL-6, IL-8 by ELISA (enzyme-linked immunosorbent assay) at 450nm. Results : In inhibiting Tyrosinase the results are as follows. 1. We observed 22% inhibition of Mushroom Tyrosinase at $500{\mu}g/m{\ell}$ concentration of GJE extracts. 2. We also could observe that the decreased Mushroom Tyrosinase activities in HCT, CIL extracts. In inhibiing $TNF-{\alpha}$, IL-6, IL-8 productions in HMC-1 cells the results are as follows. 1. Of the extracts examined, HCT, PMS, APL extracts showed over 50% inhibitions of Cytokines at $200{\mu}g/m{\ell}$ concentration. 2. In particular, APL extracts showed the best inhibitory effect on Cytokine productions in a dose-dependent manner. Conclusion : These results suggest that GJE extracts contributes to the anti melanin activities and represent a potential source of whitening agent. Thus these herbal medicines suggest novel drugs on anti-allergic effects.

• The Journal of Korean Obstetrics and Gynecology
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• v.36 no.4
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• pp.96-120
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• 2023

Objectives: This study reports the findings that support the efficacy of herbal medicine (HM) for premenstrual syndrome (PMS). Methods: We conducted meta-analysis of findings from randomized controlled trials (RCTs) for PMS treated with HM. The articles were published before July 2022, located using 9 databases (Pubmed, EMBASE, Cochrane Library, CINAHL, CNKI, CiNii, SCIENCE ON, KoreaMed, OASIS). Results: We observed 2,034 studies, of which 23 RCTs met our inclusion criteria. The risk of bias in the included studies was relatively unclear or high. Meta-analysis of 3 RCTs showed that HM group had a significantly higher total effective rate than the western medicine group (RR 1.20 [95% CI 1.06, 1.36, p=0.004]). Meta-analysis of 1 RCT showed that HM group had a significantly lower symptom score (MD -3.04 [95% CI -5.36, -0.72, p=0.01]), while there was no significant difference in daily record of severity of problems scale (MD -20.52 [95% CI -49.33, 8.29, p=0.16]). Conclusions: HM significantly improved PMS symptoms than general treatment and no serious adverse events were reported. However, the evidence on the effectiveness and safety of HM for PMS was not enough to provide reliable results due to the small number and low quality of included studies. We believe that rigorous RCTs will lead to more reliable evidence of the intervention.

• Journal of Animal Reproduction and Biotechnology
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• v.36 no.3
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• pp.121-128
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• 2021

Increasing the efficiency of HR (homologous recombination) is important for a successful knock-in. Rad51 is mainly involved in homologous recombination and is associated with strand invasion. The HR-related mismatch repair system maintains HR fidelity by heteroduplex rejection and repair. Therefore, the purpose of this study is to control Rad51, which plays a critical role in HR, through UV-induced DNA damage. It is also to confirm the effect on the expression of MMR related genes (Msh2, Msh3, Msh6, Mlh1, Pms2) and HR-related genes closely related to HR through treatment with the MMR inhibitor CdCl₂. The mRNA expression of Rad51 gene was confirmed in both HC11 cells and mouse testes, but the mRNA expression of Dmc1 gene was confirmed only in mouse testes. The protein expression of Rad51 and Dmc1 gene increased in UV-irradiated HC11 cells. After 72 hours of treatment with 1 ㎛ of CdCl₂, the mRNA expression level of Msh3, Pms2, and Rad51 decreased, but the mRNA expression level of Msh6 and Mlh1 increased in HC11 cells. There was no significant difference in Msh2 mRNA expression between CdCl₂ untreated-group and the 72 hours treated group. In conclusion, HR-related gene (Rad51) was increased by UV-induced DNA damage. Treatment of the MMR inhibitor CdCl₂ in HC11 cells decreased the mRNA expression of Rad51.

• The Journal of Korean Medicine
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• v.36 no.4
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• pp.19-34
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• 2015

Objectives: This study was performed to investigate the anti-tumor effect of O. japonicus extracts on intrahepatic cholangiocarcinoma cell line SNU-1079. Methods: Cholangiocarcinoma SNU-1079 cells were treated with various concentrations of O. japonicus DW and EtOH extracts ($0-300{\mu}g/ml$) for 24, 48 or 72 h. Cell viability was evaluated through a PMS/MTS assay, and the apoptosis rate was examined through ELISA assay and flow cytometry analysis. The mRNA expression of apoptosis- and cell cycle progression-related genes (Bcl-2, Mcl-1, Bax, Survivin, Cyclin D1, and p21) was evaluated using real-time PCR, and the caspase activity was examined using immunoblot analysis. Results: O. japonicus extracts inhibited cell proliferation and increased apoptosis rate in both ELISA assay and flow cytometry analysis. O. japonicus extracts decreased Bcl-2, Mcl-1, Survivin, and Cyclin D1 mRNA expression and increased Bax mRNA level. O. japonicus extracts also increased Caspase-3 activation. Overall, O. japonicus DW extracts were more effective than EtOH extracts. Conclusions: O. japonicus inhibited cell proliferation and induced apoptosis in SNU-1079 cells via mitochondria -mediated intrinsic pathway, which leads to Caspase-3 activation. The results indicate that O. japonicus is a potential therapeutic herb with anti-tumor effect against intrahepatic cholangiocarcinoma.

• Korean Journal of Animal Reproduction
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• v.4 no.1
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• pp.75-82
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• 1980

This experiment was carried out to improve a simple and effective procedure of egg transfer which is considered to be the most useful technique for the improvementand proliferation of domestic animals. Several experiment procedures such as superovulation, surgical recovery of ovulated egg, in vitro capacitation of ejaculated spermatozoa, in vitro fertilization and culture of embryo were conducted. The results obtained in this experiment were summarized as follows: 1. In rabbits treated with PMS in combination with estradiol and HCG, 10 to 43 eggs were obtained in one rabbit and the average ovulation number of 20 rabbits was 21 eggs. 2. Six to 24 eggs (average 13 eggs) were recovered from the removed oviducts by the methods of flushing technique and the average recovery rate of 20 rabbits was 61.9 percent. 3. Most rabbit spermatozoa ejaculated by artificial vagina were capacitated in vitro by the culture of the spermatozoa with PBI medium at 37$^{\circ}C$ for 4 hours after removal of seminal plasma. 4. Normal fetilization following in vitro fetilization was observed in 88 (42.7%) of 206 eggs. 5. The number of eggs developed to 2-, 4-, and 8-cell stage following in vitro culture with PBI medium at 37$^{\circ}C$ was 26 (70.3%), 20 (54.1%) and 17 (45.9%) of 37 eggs used for in vitro culture.

• The Journal of Korean Medicine
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• v.41 no.4
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• pp.12-26
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• 2020

Objectives: The aim of this study is to investigate the mechanisms involved in the anti-inflammatory and anti-tumor effects of Rhus verniciflua Stokes (RVS) on PMA-differentiated human monocytic leukemia THP-1 cells. Methods: Cells were treated with various concentrations of RVS decoction (0-300㎍/ml) for 24, 48, and 72h. Cell viability was evaluated by MTS/PMS assay. The expressions of MMP-2, MMP-9, TIMP-1, TIMP-2, iNOS and COX-2 mRNA and proteins were measured using RT-PCR and western blotting, respectively. Results: RVS suppressed expression of MMP-2 and MMP-9 mRNA. It also down-regulated iNOS and COX-2 mRNA and protein expression. RVS inhibited NF-𝜅B p65 activity and the phosphorylation of Akt and MAPK (ERK and p38 MAPK). Instead, the phosphorylation of JNK is increased at a very low concentration but decreased at higher concentrations. Conclusion: RVS is regarded to inhibit the expression of MMP and TIMP as well as iNOS and COX-2 gene expression via directly inhibiting the activation of NF-𝜅B and phosphorylation of MAPK pathway in THP-1 cells. This suggests RVS have potential to be used as a therapeutic agent for acute myeloid leukemia (AML).

• Journal of Animal Reproduction and Biotechnology
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• v.38 no.3
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• pp.99-108
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• 2023

Background: Efficient gene editing technology is needed for successful knock-in. Homologous recombination (HR) is a major double-strand break repair pathway that can be utilized for accurately inserting foreign genes into the genome. HR occurs during the S/G2 phase, and the DNA mismatch repair (MMR) pathway is inextricably linked to HR to maintain HR fidelity. This study was conducted to investigate the effect of inhibiting MMR-related genes using CdCl₂, an MMR-related gene inhibitor, on HR efficiency in HC11 cells. Methods: The mRNA and protein expression levels of MMR-related genes (Msh2, Msh3, Msh6, Mlh1, Pms2), the HR-related gene Rad51, and the NHEJ-related gene DNA Ligase IV were assessed in HC11 cells treated with 10 μM of CdCl₂ for 48 hours. In addition, HC11 cells were transfected with a CRISPR/sgRNA expression vector and a knock-in vector targeting Exon3 of the mouse-beta casein locus, and treated with 10 μM cadmium for 48 hours. The knock-in efficiency was monitored through PCR. Results: The treatment of HC11 cells with a high-dose of CdCl₂ decreased the mRNA expression of the HR-related gene Rad51 in HC11 cells. In addition, the inhibition of MMR-related genes through CdCl₂ treatment did not lead to an increase in knock-in efficiency. Conclusions: The inhibition of MMR-related gene expression through high-dose CdCl₂ treatment reduces the expression of the HR-related gene Rad51, which is active during recombination. Therefore, it was determined that CdCl₂ is an inappropriate compound for improving HR efficiency.

• The Journal of Korean Medicine
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• v.40 no.2
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• pp.35-50
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• 2019

Objectives: Orostachys japonicas (O. japonicus) has been known for its anti-tumor effect. In the present study, it was investigated whether O. japonicus EtOH extracts could induce apoptosis and autophagy which are part of the main mechanism related to anti-tumor effect in THP-1 cells. Methods: Cells were treated with various concentrations of O. japonicus EtOH extracts ($0-300{\mu}g/ml$) for 24, 48, and 72h. Cell viability was evaluated by MTS/PMS assay and apoptosis rate was examined by flow cytometry and ELISA assay. The mRNA expression of apoptosis-related genes (Bcl-2, Mcl-1, Survivin, Bax) and autophagy-related gene (mTOR) was evaluated using real-time PCR. The protein expression of Caspase-3, Akt, LC3 II, Beclin-1, Atg5, $NF-{\kappa}B$, p38, ERK was evaluated using western blot analysis. Results: O. japonicus EtOH extracts inhibited cell proliferation and apoptosis rate was increased in both flow cytometry and ELISA assay. Bcl-2, Mcl-1, Survivin (anti-apoptosis factors) mRNA expressions were decreased and Bax (pro-apoptosis factor) mRNA level was increased. mTOR mRNA expressions was decreased and LC3 II protein expressions was increased. Activation of $NF-{\kappa}B$ was decreased and phosphorylation of p38 was increased. Conclusion: O. japonicus is regarded to inhibit cell proliferation, to induce apoptosis and to regulate autophagy-related genes in THP-1 cells via $NF-{\kappa}B$ and p38 MAPK signaling pathway. This suggests O. japonicus could be an effective herb in treating acute myeloid leukemia.

• The Journal of Internal Korean Medicine
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• v.32 no.2
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• pp.153-169
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• 2011

Objectives : This study was performed to investigate the anti-fibrogenic effect and the effect on cell growth and apoptosis in YJCGT, YJCGT YSO and YJCGT YSCO on thioacetamide-induced rat liver tissue and the immortalized human hepatic cell line LX2. Materials and Methods : LX2 cells were treated with various concentrations (0, 50, 150, 300 ug/ml) of YJCGT, Y+YSO, and Y+YSCO extract for 24, 48 and 72 hours. After the treatment, cell viability was measured by using MTT assay. Caspase inhibitor assay, and cell viability were determined by a colorimetric assay with PMS/MTS solution. Rat liver fibrosis was induced by intraperitoneal thioacetamide injection 150 mg/kg 3 times a week for 5 weeks. After the treatment, body weight, liver & spleen weights, liver function test, the complete blood cell count and the change of portal pressure were studied. After YJCGT, Y+YSO, and Y+YSCO treatment, percentages of collagen in thioacetamide-induced rat liver tissue were measured. Results : The viability of the LX2 cell decreased in a dose- and time-dependent manner. Exposure of LX2 cells to YJCGT, YJCGT+YSO and YJCGT+YSCO induced caspase-3 activation, but co-treatment of YJCGT, YJCGT+YSO and YJCGT+YSCO with the pan-caspase inhibitor Z-VAD-FMK, and the caspase-3 inhibitor Z-DEVE-FMK, blocked apoptosis. There was no difference in rat body weight between the thioacetamide only group and the YJCGT, YJCGT+YSO and YJCGT+YSCO groups. In the YJCGT, YJCGT YSO and YJCGT YSCO groups, the serum level of GPT significantly went down compared with the thioacetamide only group. In the YJCGT, Y+YSO, Y+YSCO groups, white blood cell elevated by thioacetamide injection decreased but RBC, Hgb, and Hct increased. In the Y+YSO group, the portal pressure elevated by thioacetamide injection significantly decreased. In the histological finding, thioacetamide injections caused severe fibrosis, but YJCGT, Y+YSO, and Y+YSCO treatment significantly reduced the amounts of hepatic collagens. Conclusions : YJCGT, Y+YSO, and Y+YSCO inhibit the growth of LX2 cells by inducing apoptosis through caspase activity. YJCGT, Y+YSO, and Y+YSCO have beneficial effects on the treatment of cirrhotic patients as well as patients with chronic hepatitis.