• International Journal of Oral Biology
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• 제38권3호
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• pp.101-110
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• 2013

We investigated the synergistic apoptotic effects of co-treatments with Chios gum mastic (CGM) and eugenol on G361 human melanoma cells. An MTT assay was conducted to investigate whether this co-treatment efficiently reduces the viability of G361 cells compared with each single treatment. The induction and augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining, and analyses of DNA hypoploidy. Western blot analysis and immunofluorescent staining were also performed to evaluate expression and translocation of apoptosis-related proteins following CGM and eugenol co-treatment. Proteasome activity and mitochondrial membrane potential (MMP) changes were also assayed.The results indicated that the co-treatment of CGM and eugenol induces multiple pathways and processes associated with an apoptotic response in G361 cells. These include nuclear condensation, DNA fragmentation, a reduction in MMP and proteasome activity, an increase of Bax and decrease of Bcl-2, a decreased DNA content, cytochrome c release into the cytosol, the translocation of AIF and DFF40 (CAD) into the nucleus, and the activation of caspase-9, caspase-7, caspase-3, PARP and DFF45 (ICAD). In contrast, separate treatments of $40{\mu}g/ml$ CGM or $300{\mu}M$ eugenol for 24 hours did not induce apoptosis. Our present data thus suggest that a combination therapy of CGM and eugenol is a potential treatment strategy for human melanoma.

• Biomolecules & Therapeutics
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• 제20권3호
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• pp.293-298
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• 2012

The purpose of this study was to investigate the modification of expression and functionality of the drug transporter P-glycoprotein (P-gp) by tumor necrosis factor-alpha (TNF-${\alpha}$) and interferon-gamma (IFN-${\gamma}$) at the blood-brain barrier (BBB). We used immortalized human brain microvessel endothelial cells (iHBMEC) and primary human brain microvessel endothelial cells (pHBMEC) as in vitro BBB model. To investigate the change of p-gp expression, we carried out real time PCR analysis and Western blotting. To test the change of p-gp activity, we performed rhodamin123 (Rh123) accumulation study in the cells. In results of real time PCR analysis, the P-gp mRNA expression was increased by TNF-${\alpha}$ or IFN-${\gamma}$ treatment for 24 hr in both cell types. However, 48 hr treatment of TNF-${\alpha}$ or IFN-${\gamma}$ did not affect P-gp mRNA expression. In addition, co-treatment of TNF-${\alpha}$ and IFN-${\gamma}$ markedly increased the P-gp mRNA expression in both cells. TNF-${\alpha}$ or IFN-${\gamma}$ did not influence P-gp protein expression whatever the concentration of cytokines or duration of treatment in both cells. However, P-gp expression was increased after treatments of both cytokines together in iHBMEC cells only compared with untreated control. Furthermore, in both cell lines, TNF-${\alpha}$ or IFN-${\gamma}$ induced significant decrease of P-gp activity for 24 hr treatment. And, both cytokines combination treatment also decreased significantly P-gp activity. These results suggest that P-gp expression and function at the BBB is modulated by TNF-${\alpha}$ or/and IFN-${\gamma}$. Therefore, the distribution of P-gp depending drugs in the central nervous system can be modulated by neurological inflammatory diseases.

• 대한한의학회지
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• 제24권2호
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• pp.138-147
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• 2003

Objectives : In this study, we investigated the mechanism by which Chelidonium majus (CM) regulates nitric oxide (NO) production. Methods : Using mouse peritoneal macrophages, the mechanism by which CM regulates NO or tumor necrosis $factor-{\alpha}(TNF-{\alpha})$ production was examined. NO release was measured by the Griess method. $TNF-{\alpha}$ production was measured by the ELISA method. The protein extracts were prepared and samples were analyzed for the inducible NOS(iNOS) expression and nuclear factor kappa $B(NF-{\kappa}B)$ activation by Western blotting. Results : When CM was used in combination with recombinant $interferon-{\gamma}{\;}(rIFN-{\gamma})$, there was a marked cooperative induction of NO production. CM had an effect on NO production by itself. The expression of the iNOS gene was increased in $rIFN-{\gamma}$ plus CM-stimulated peritoneal macrophages and almost completely inhibited by pre-treatment with pyrrolidine dithiocarbamate (PDTC), an inhibitor of $NF-{\kappa}B$. The $NF-{\kappa}B$ activation was increased in rIFN-{\gamma} plus CM-induced peritoneal macrophages. The increased production of NO from $rIFN-{\gamma}$ plus CM-stimulated peritoneal rnacrophages was decreased by the treatment with $N^{G}-monomethyl-{_L}-arginine{\;}(N^{G}MMA){\;}N^{\alpha}-Tosyl-Phe$ chloromethyl ketone (TPCK) , and was almost completely inhibited by pre-treatment with PDTC. Furthermore, treatment with CM alone or rIFN-{\gamma} plus CM in peritoneal macrophages caused a significant increase in $TNF-{\alpha}$ production. PDTC decreased CM-induced $TNF-{\alpha}$ production significantly. After CM treatment in HT-29 or AGS cells, cell viability decreased. Conclusions : These findings demonstrate that CM increases the production of NO and $TNF-{\alpha}{\;}by{\;}rIFN-{\gamma}-primed$ macrophages and suggest that NF-B plays a critical role in mediating these effects of CM.

• Tuberculosis and Respiratory Diseases
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• 제62권1호
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• pp.33-42
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• 2007

연구배경: 다양한 고형암에서 HO-1의 높은 발현이 알려져 있고, 그것의 항산화와 항세포고사의 역할로 인해 빠른 암종의 성장에 중요한 역할이 있음이 보고되고 있다. 대표적인 활성산소종 생성 항암제인 Cisplatin은 현재까지 폐암치료에 가장 광범위하게 사용되고 있으나, 여러 내성발생이 임상치료의 주요문제로 대두되고 있다. 저자들은 A549 폐암세포주에서 HO-1의 발현이 증가되었고 HO-1 활성억제제나 siRNA 방법을 통해 생존율의 의미 있는 감소와 세포고사가 유도됨을 보고한 바 있다. 이 연구의 목적은 A549 폐암세포주에 cisplatin 처리시 HO-1의 발현의 증가유무와 기전을 규명하고 실제 HO-1의 억제가 cisplatin에 의한 항암제 감수성을 증가시키는지를 알아보는데 있다. 방 법: 비소세포폐암세포주인 A549, NCI-H23, NCIH157, NCI-H460을 이용하였다. 세포독성은 MTT 방법으로 구하였고, HO-1, Nrf2, MAPK의 발현은 Western blotting으로 확인하였다. 또한 MAPK억제제들을 전처치한 후 cisplatin에 의해 유도된 Nrf2와 HO-1의 발현에 미치는 영향을 역시 Western blotting으로 관찰하였다. A549세포에 활성억제제인 ZnPP나 HO-1 small interfering RNA (siRNA)을 주입하여 cisplatin과의 병합요법시 생존율의 배가효과 유무를 MTT 방법으로 확인하였고, 이러한 효과가 ROS 형성과 HO-1의 발현변화와 관련되는지를 알아보기 위해 $carboxy-H_2DCFDA$ 방법과 Western blotting을 통해 각각 확인하였다. 결 과: Cisplatin 처리시 다른 세포주에 비해 A549 세포가 의의 있게 내성을 보였다. $10{\mu}M$의 농도에서 시간 의존적으로 HO-1, Nrf2, MAPK의 발현이 증가하였고, MAPK 억제제들을 전 처치하였을 때 cisplatin에 의해 유도된 HO-1과 Nrf2의 발현이 억제됨을 확인하였다. HO-1의 활성억제제인 ZnPP와 HO-1 siRNA를 통해 HO-1 mRNA를 직접 억제하는 방법으로 cisplatin과 병합치료시 단독치료에 비해 의의 있는 생존율의 감소를 보였다. 이러한 효과는 활성산소종의 생성 증가와 HO-1의 발현억제에 의한 결과임을 확인하였다. 결 론: Cisplatin 처리시 HO-1의 발현은 MAPKNrf2-HO-1의 경로를 통해 증가하였고, 부분적으로 치료에 대한 내성과 관련이 있었으며, ZnPP 등의 활성억제제나 siRNA를 통한 knock-down 방법으로 HO-1 을 표적으로 억제하는 치료방법을 통해 cisplatin의 항암제 감수성을 증가시켰다.

• 사상체질의학회지
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• 제25권4호
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• pp.432-441
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• 2013

Objectives Despite the treatment with antibiotics, patients with sepsis has a high mortality (80%) in the underlying disease group. The aim of this study was to report the improvement of septic condition of the cholangiocarcinoma patient after the treatment with Handayeolso-tang, Fel Tauri, and antibiotics. Methods We retrospectively reviewed the medical records. The patient's subjective symptoms such as chilling and abdominal pain were evaluated by NRS and the performance status was evaluated by ECOG. This case was literally compared with relevant published studies on prognosis of sepsis. Results Despite poor prognostic factor(MEDS score 18), the patient's symptoms such as fever, chilling, abdominal pain, and diarrhea and ECOG(Eastern Cooperative Oncology Group) improved. The patient was hemodynamically stabilized on 3rd day from the treatment, and her laboratory test results were normalized on 7th day. Conclusions A female patient of metastatic cholangiocarcinoma came to the hospital for cholangitis, later causing septic shock. Both her symptoms and laboratory tests showed significant improvement after the treatment of antibiotics, Handayeolso-tang and Fel Tauri. To our knowledge, this is the first case reporting the synergistic combination of Korean oriental medicine and Western medicine approaching to sepsis.

• 대한한방내과학회지
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• 제18권1호
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• pp.207-217
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• 1997

The oriental medicine based on the traditional Chinese medicine has developed characteristically according to the history and racial character respectively; China, Korea and Japan. Japan, among these nations, has accepted western medicine earlier than other nations and has tried to compare western and oriental medicine and combine them. In Japanese traditional medicine, it is characteristic that the old medical classics focusing on Sanghannon (傷寒論) and Geumgyeyoryak(金?要略) has developed The recent tendencies of clinical medicine and researches in Korean oriental medicine are mostly about the study of oriental medicine in view of western medicine and the combination of western and oriental medical treatment like Japan. But the study on the Japanese oriental medicine hasn't so far been tried before in Korea. From now on, we should not overlook that a more interest on Japanese oriental medicine will be very useful. Therefore we have surveyed the background of its origin and the process of development of the theory of ${\ulcorner}$Qi, Blood and Body Fluids${\lrcorner}$. What we wish to show in this paper is to provide a source for the basic understanding by explaining a fundamental theory of physiology and pathology of Japanese oriental medicine. Concepts of ${\ulcorner}$Qi, Blood and Body Fluids${\lrcorner}$ suggested by Nangai Yoshimashi in 1792 is the way of thinking that the circulation of 3 factors- ${\ulcorner}$Qi, Blood and Body Fluids${\lrcorner}$ nourish human body. Among these 3 factors, if Qi does not function smoothly, it causes the condition of a disease like Qi-deficiency, imbalance of Qi-distribution or Qi-depression and stasis; in Blood's case, deficiency of Blood and Blood stasis; and as for Body Fluids, stasis of Body Fluids. In the recent trend of study, there's a try to combining the western and oriental medicine, Qi is considered as psychoneurotic system, Blood as circulatory and endocrinologic system and Body Fluids as immunologic system.

• 대한한의학방제학회지
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• 제27권1호
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• pp.45-52
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• 2019

Objectives : Hyperthermia is a widely used therapeutic tool for cancer therapy and a well-known inducer of apoptosis. Although the Cinnamomi cortex (CC) is a potent anticancer agent for several human carcinomas, it is less potent in the human U937 cell line. To explore any enhancing effects of CC with hyperthermia induced apoptosis, this study investigated the combined effects and apoptotic mechanisms of hyperthermia and CC in U937 cells. Methods : U937 cells were heat treated at $43^{\circ}C$ for 30 min with or without pre-treatment for 1h with CC and then incubated at $37^{\circ}C$ with 5% $CO_2$. Cell viability was analyzed by MTT assay and Trypan blue assay. Morphological changes reflecting apoptosis were visualized under microscope. Synergy effect of CC combined with hyperthermia were calculated by Compusyn software. The expression of proteins related to apoptosis and signaling pathways was determined by western blotting. Results : Hyperthermia with CC reduced cell viability and induced apoptosis. Combined hyperthermia and CC treatment markedly augmented apoptosis by upregulating proapoptotic proteins and suppressing antiapoptotic proteins, culminating in caspase-3 activation. Furthermore, the combined treatment, decreased the expression of in Bcl-2 family, cyclin D1, VEGF, MMP2 and MMP9 expression. Conclusion : This study provides compelling evidence that hyperthermia, in combination with CC, is a promising therapeutic strategy for enhancement of apoptosis and suggests a promising therapeutic approach for cancer.

• 한방재활의학과학회지
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• 제27권3호
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• pp.137-146
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• 2017

Objectives The purpose of this study is to compare the effects between the KL-Grade and improvement of knee pain treated by Korean Medicine therapy. Methods 114 patients who received inpatient treatment from July 2014 to May 2017 in the Daejeon Jaseng of Korean Medicine Hospital were divided into 5 groups by the KL-Grade. All patients received a combination of treatment including acupunture, pharmacopunture, herbal medication. They were compared and analyzed on the basis of improvement between measuring Numeric Rating Scale (NRS), Western Ontario and McMaster Universities Arthritis Index (WOMAC Index), EuroQol-5 Dimension Index (EQ-5D Index) as they were hospitalized and as they were discharged. The statistically significance was evaluated by SPSS 23.0 for windows. Results After treatment, KL-Grade 0 group's Numeric Rating Scale (NRS), Western Ontario and McMaster Universities Arthritis Index (WOMAC Index), EuroQol-5 Dimension Index (EQ-5D Index) improvement was $2.02{\pm}1.69$, $7.50{\pm}9.67$ and $0.11{\pm}0.15$ respectively. KL-Grade 1 group's improvement was $2.09{\pm}1.23$, $11.75{\pm}13.99$ and $0.12{\pm}0.13$ respectively. KL-Grade 2 group's improvement was $1.60{\pm}1.07$ and $14.70{\pm}14.19$ respectively. But In this group, EQ-5D Index has decreased by $0.01{\pm}0.10$. KL-Grade 3 group's improvement was $1.88{\pm}1.31$, $7.81{\pm}13.35$ and $0.13{\pm}0.20$ respectively (p<0.034). In the case of KL-Grade 4, the population was not statistically significant (N=2) and therefore excluded from statistical significance. And there was no statistically significance between 4 group's improvement after treatment (p>0.05). Conclusions The above study showed that Korean medicine treatments showed significant therapeutic effects on knee pain and degenerative knee joints, but there was no significant difference in the effectiveness of degenerative arthritis (KL-Grade).

• Molecules and Cells
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• 제38권7호
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• pp.638-642
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• 2015

Quercetin can suppress osteosarcoma cell growth and metastasis. However, other effects of quercetin on osteosarcoma remain largely unknown. This research aims to evaluate the effects of quercetin in combination with cisplatin as treatment for osteosarcoma and investigate its regulatory mechanism. Cell viability and apoptosis in 143B cell line were determined after treatment with quercetin and/or cisplatin. RT-PCR and Western blot analysis were performed to determine the RNA or protein expression levels. Moreover, transwell assay was used to evaluate metastasis. Furthermore, rescue experiments were performed to investigate the potential regulatory mechanism of the treatment. Results showed that quercetin with concentration that was equal to or greater than $10{\mu}M$ inhibited 143B proliferation, while $5{\mu}M$ quercetin enhanced the cisplatin sensitivity of 143B cells. Expression of miR-217 was upregulated after quercetin and/or cisplatin treatment, while its target KRAS was downregulated both at mRNA and protein levels. MiR-217 knockdown led to the loss of enhanced cisplatin sensitivity while miR-217 overexpression showed the opposite effects, indicating that quercetin regulated cisplatin sensitivity by modulating the miR-217-KRAS axis. In conclusion, $5{\mu}M$ quercetin enhanced the cisplatin sensitivity by modulating the miR-217-KRAS axis. This finding suggests that quercetin may be administered with cisplatin to improve the treatment for osteosarcoma.

• 대한한방내과학회지
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• 제36권1호
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• pp.58-68
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• 2015

목 적: 소세포 폐암 및 전이성 폐암은 폐암 중에서도 가장 예후가 불량하고 생존율도 상대적으로 낮은 암으로 알려져 있다. 난치성 폐암 환자에 대해 한방치료가 생존 기간 연장 및 삶의 질 관리면에 있어서 효과가 있음을 보여주고자 한다. 연구방법 및 대상: 본원에서 입원 및 통원치료를 병행한 소세포폐암 환자 1명과 원발성 간암에서 폐로 전이된 전이성 폐암 환자 1명에 대하여 한방치료의 효과 및 임상경과를 후향적으로 조사하였다. 치료기간은 각각 2000년 1월-2009년 12월과 2004년 9월-2014년 2월이었으며 한약치료는 평균 1개월 간격으로 행해졌으며 입원기간 동안에는 한약치료를 포함한 침구치료를 추가로 시행하였다. 치료효과 및 경과 판정을 위해 흉부 방사선 검사 및 혈액검사를 평균 1개월 간격으로 시행하였으며 내원시마다 환자의 증상 및 상태를 확인하였다. 결 과: 2명의 폐암 환자 모두 꾸준한 한방치료를 받으며 진단시점부터 9년 이상의 상당히 오랜 기간 동안 비교적 좋은 삶의 질을 유지하면서 종양으로 인한 임상경과 또한 완만하게 진행이 되었다. 결 론: 본 증례는 한방치료가 불응성 폐암 환자에 대해 삶의 질을 양호하게 유지하고 증상 조절 및 종양의 진행양상을 완화시켜 주며 나아가 생존기간 연장에도 효과가 있음을 보여준다.