• 한국미생물·생명공학회지
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• 제18권5호
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• pp.525-529
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• 1990

식물세포인 Lithospermum erythrorhizon을 교반 반응기와 calcium alghinate에 고정화된 상태로 충전층 반응기에서 n-hexadecane으로 동시 추출하면서 shikonin을 생산하여 각각의 생산성을 비교하였다. 교반 반응기에서 shikonin의 비생산과 부피생산성은 각각 1.5mg shikonin/g cell과 400$\mu g$ shikonin/(L.day)였고 충전층 반응기에서는 가각 2.0mg shikonin/g cell과 2857 $\mu g$ shikonin/(L.day)였으며 이는 각각 교반 반응기에 비하여 1.3, 7.1배 높은 것이다. 충전층 반응기에서 shikonin의 생산성이 높은 것은 calcium alginate 입자에 세포가 고농도고 축적되어 단위 반응기 부피당 세로의 부하 능력이 높고 또한 세포가 서로 접촉하기가 쉽고 고정화 입자내의 환경이 세포가 분화하기에 좋은 조건을 형성하기 때문인 것으로 사료된다.

• 동의생리병리학회지
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• 제19권5호
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• pp.1363-1369
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• 2005

Previously we have shown that shikonin has strong anti-tumor activities via inducing apoptosis and suppressing metastasis on LLC cells in vivo and in vitro. Here we have investigated anti-angiogenic potential of shikonin and its possible mechanism of action in HSE cells. Shikonin inhibited the proliferation of HSE cells in a concentration-dependent manner. It was shown that this proliferation inhibition was caused by apoptosis induced by shikonin via BrdU incorporation and Western blotting analysis. Shikonin treatment was caused that decrease of activation of caspases and cleavage of PARP. And shikonin induced that the activation of mitogen-activated protein kinases (MAPKs), such as extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38. Moreover, shikonin showed anti-angiogenic activities inhibiting tube-like formation of HSE cells in vitro and vascular formation of LLC cells in vivo. These findings suggest that shikonin may a possible candidate not only anti-metastatic agent but also anti-angiogenic agent.

• 동의생리병리학회지
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• 제19권2호
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• pp.501-507
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• 2005

Shikonin is a chemically characterized component of traditional herbal medicine, the root of Lithospermum erythrorhizon and has been shown to possess antitumor activities. Here we investigated anticancer potential of shikonin and its possible mechanism of action in LLC cells. Shikonin inhibited the proliferation of LLC cells in a concentration-dependent manner. It was also demonstrated that shikonin induced apoptosis in LLC cells by Annexin V staining and TUNEL staining analysis. Shikonin treatment was caused that decrease of Bcl-2, activation of caspases and cleavage of PARP. And shikonin also induced that the activation of mitogen-activated protein kinases (MAPKs), such as extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38. Interestingly, the cell proliferation inhibition induced by shikonin was recovered by specific inhibitors of JNK and p38 but the inhibitor of MEK, the upstream kinase of ERK, did not recover. Additionally, shikonin administration at doses of 5 mg/kg in C57BL/6 mice strongly inhibited the primary tumor growth of LLC. Taken together, these results suggest that shikonin may suppress LLC cell proliferation by inducing an apoptotic process via activation of caspases and MAPKs

• Biomolecules & Therapeutics
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• 제30권2호
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• pp.137-144
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• 2022

Radiation resistance represents an imperative obstacle in the treatment of patients with colorectal cancer, which remains difficult to overcome. Here, we explored the anti-proliferative and migration-inhibiting properties of the natural product shikonin on a radiation-resistant human colon carcinoma cell line (SNU-C5RR). Shikonin reduced the viability of these cells in a dose-dependent manner; 38 µM of shikonin was determined as the half-maximal inhibitory concentration. Shikonin induced apoptotic cell death, as demonstrated by increased apoptotic body formation and the number of TUNEL-positive cells. Moreover, shikonin enhanced mitochondrial membrane depolarization and Bax expression and also decreased Bcl-2 expression with translocation of cytochrome c from mitochondria into the cytosol. In addition, shikonin activated mitogen-activated protein kinases, and their specific inhibitors reduced the cytotoxic effects of shikonin. Additionally, shikonin decreased the migration of SNU-C5RR cells via the upregulation of E-cadherin and downregulation of N-cadherin. Taken together, these results suggest that shikonin induces mitochondria-mediated apoptosis and attenuates epithelial-mesenchymal transition in SNU-C5RR cells.

• The Korean Journal of Physiology and Pharmacology
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• 제27권4호
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• pp.357-364
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• 2023

Sjögren syndrome (SS) is a systemic inflammatory autoimmune disease that involves exocrine glands. Shikonin is extracted from comfrey, which is conventionally used as an anti-tumor, antibacterial, and antiviral drug in China. However, the application of Shikonin in SS remains unreported. This study aimed to verify the potential functions of Shikonin in SS progression. Firstly, non-obese diabetic mice were used as the SS mouse model, with C57BL/6 mice serving as the healthy control. It was demonstrated that the salivary gland damage and inflammation were aggravated in the SS mouse model. Shikonin improved salivary gland function decline and injury in the SS mouse model. Moreover, Shikonin reduced inflammatory cytokines and immune infiltration in the SS mouse model. Further experiments discovered that Shikonin attenuated the MAPK signaling pathway in the SS mouse model. Lastly, inhibition of the MAPK signaling pathway combined with Shikonin treatment further alleviated the symptoms of SS. In conclusion, Shikonin ameliorated salivary gland damage and inflammation in a mouse model of SS by modulating the MAPK signaling pathway. Our findings indicate that Shikonin may be a useful drug for SS treatment.

• 한국미생물·생명공학회지
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• 제17권4호
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• pp.343-348
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• 1989

L. erythrorhizon 세포를 polyurethane foam과 함께 증식시킬 경우 shikonin 유도체가 polyurethane에 효과적으로 흡착됨과 동시에 polyurethane을 사용하지 않은 경우와 비교하여 shikonin 생산량이 현저히 증가하였다. 이 같은 증가는 세포를 polyurethane pore에 고정하여 증식시킴으로써 원활한 세포간 접촉을 유지하고 세포 내에 shikonin 농도를 저하시켜shikonin 생성에 좋은 조건을 제공함에 기인한 것으로 생각되었다. 공정의 생산성을 높이기 위하여 여러가지 배양시스템이 검토되었는데, indole-3-acetic acid(1.75mg/ι)와 kinetin(0.1mg/ι)을 함유하는 Schenk-Hildebrandt 배지 (SHIK 배지) 시스템이 가장 효과적이었다. p-Chlorophenoxyacetic acid (2.0 mg/ι)와 kinetin (0.1 mg/ι)를 함유하는 Schenk-Hildebrandt 배지 (SHND 배지) 시스템에 비교하여 SHIK 배지 시스템에서 Shikonin 생성량은 약 4.5배 증가하였다. Polyurethane을 세포를 고정화하는 지지체로 사용할 경우에는 현재 행하여지고 있는 2단계 배양보다 1단계 배양이 더욱 효과적이며 경제적으로도 매우 유리할 것으로 판단되었다.

• The Korean Journal of Physiology and Pharmacology
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• 제14권4호
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• pp.199-204
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• 2010

The potential therapeutic action of shikonin in an experimental model of rheumatoid arthritis (RA) was investigated. As a RA animal model, DBA/1J mice were immunized two times with type II collagen. After the second collagen immunization, mice were orally administered shikonin (2 mg/kg) once a day for 35 days, and the incidence, clinical score, bone mineral density (BMD), bone mineral content (BMC) and joint histopathology were evaluated. BMD in the proximal regions of the tibia largely increased in the shikonin treatment group compared with the control group. We also examined the effect of shikonin on inflammatory cytokines and cartilage protection. Shikonin treatment significantly reduced the incidence and severity of collagen-induced arthritis (CIA), markedly abrogating joint swelling and cartilage destruction. Shikonin also significantly inhibited the production of matrix metalloproteinase (MMP)-1 and up-regulated tissue inhibitors of metalloproteinase (TIMP)-1 in mice with CIA. In conclusion, shikonin exerted therapeutic effects through regulation of MMP/TIMP; these results suggest that shikonin is an outstanding candidate as a cartilage protective medicine for RA.

• 약학회지
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• 제24권3_4호
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• pp.151-157
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• 1980

Color transition mechanism of shikonin as an acid-alkali indicator was studied. It was confirmed that the presence of phenolic hydroxy radical was essential for the color change of shikonin. But in accordance with shikonin sodium salt (blue color), which was presumed to make chelation as six membered rings. Shikonin in alkaline solution, by dissociated phenolic protons of naphthoquinone nucleous, converted to the corresponding anion and instead of disappearance tautomerization, electron delocalization occurred and an additional pair of nonbonding electrons in the anion was available for interaction with .phi. electron system of the ring with further extension of the conjugation. It was responsible for its blue color(corresponding color: orange) with needs less energy difference (${\phi}{\rarw}{\phi}^{*}$) because of conjugation extension. Shikonin sodium salt seems to have similar nuclear structure as shikonin anion.

• 생명과학회지
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• 제19권11호
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• pp.1637-1643
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• 2009

천연생약 자초의 한 성분인 shikonin은 항염증, 항암 및 항비만 등 다양한 분야에 효과를 보여왔다. 이번 연구에서는 shikonin이유도하는 adipogenesis억제 과정에 어떤 인자들이 작용하는지 살펴보았다. Shikonin의 효과에 대한 분자적 메커니즘을 규명하기 위해, real-time PCR을 이용하여 C/EBPs, $PPAR{\gamma}$를 포함한 다양한 adipogenesis 인자들의 mRNA 발현량을 분석하였다. 그 결과, 초기 분화의 주요 조절자인 C/$EBP{\beta}$와 C/$EPB{\delta}$는 shikonin에 의해 거의 변화가 없었으나, 후기 분화의 주요 조절자인 $PPAR{\gamma}$와 C/$EPB{\alpha}$의 mRNA 발현은 유의하게 감소하였다. Shikonin에 의한 adipogenesis억제의 메커니즘을 좀 더 자세히 밝히기 위해 adipogenesis과정의 상위 단계에 위치한 조절자들의 mRNA 발현을 분석하였다. C/$EBP{\beta}$의 상위 조절자인 C/$EBP{\gamma}$, CHOP은 shikonin에 의해 영향을 받지 않았으나, KROX20의 경우 유의하게 감소하였다. 이러한 결과는 Pro-adipogenic 인자인 KROX20의 감소가 C/$EBP{\beta}$에 영향을 주기 보다는 C/$EBP{\beta}$와 독립적으로 그 하위의 인자들에게 영향을 줄 수 있음을 제시한다. $PPAR{\gamma}$의 상위 조절자로 알려져 있는 KLF 들 중에서 pro-adipogenic 인자인 KLF15의 mRNA 발현은 shikonin에 의해 급격히 감소하였으나 anti-adipogenic 인자인 KLF2는 shikonin에 의한 변화가 거의 없었다. 또 다른 pro-adipogenic 인자인 KLF5의 경우, 주로 작용하는 초기 분화에서는 shikonin에 의해 거의 변화가 없었지만, 후기 분화에서는 조금 증가하였다. 이러한 후기 분화에서의 KLF5의 변화는 KLF15에 비해 전체 분화에 크게 영향을 주지 못하는 것 같다. 결론적으로, shikonin은 pro-adipogenic 인자인, KROX20과 KLF15의 조절을 통해 $PPAR{\gamma}$ 및 C/$EPB{\alpha}$의 mRNA 발현을 억제함으로써 지방 세포의 분화를 저해한다고 사료된다.

• International Journal of Oral Biology
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• 제40권1호
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• pp.51-61
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• 2015

Shikonin, a major ingredient in the traditional Chinese herb Lithospermumerythrorhizon, exhibits multiple biological functions including antimicrobial, anti-inflammatory, and antitumor effects. It has recently been reported that shikonin displays antitumor properties in many cancers. This study was aimed to investigate whether shikonin could inhibit oral squamous carcinoma cell (OSCC) growth via mechanisms of apoptosis and cell cycle arrest. The effects of shikonin on the viability and growth of OSCC cell line, SCC25 cells were assessed by MTT assay and clonogenic assays, respectively. Hoechst staining and DNA electrophoresis indicated that the shikonin-treated SCC25 cells were undergoing apoptosis. Western blotting, immunocytochemistry, confocal microscopy, flow cytometry, MMP activity, and proteasome activity also supported the finding that shikonin induces apoptosis. Shikonin treatment of SCC25 cells resulted in a time- and dose-dependent decrease in cell viability, inhibition of cell growth, and increase in apoptotic cell death. The treated SCC25 cells showed several lines of apoptotic manifestation as follows: nuclear condensation; DNA fragmentation; reduced MMP and proteasome activity; decrease in DNA contents; release of cytochrome c into cytosol; translocation of AIF and DFF40 (CAD) onto the nuclei; a significant shift in Bax/Bcl-2 ratio; and activation of caspase-9, -7, -6, and -3, as well as PARP, lamin A/C, and DFF45 (ICAD). Shikonin treatment also resulted in down-regulation of the G1 cell cycle-related proteins and up-regulation of $p27^{KIP1}$. Taken together, our present findings demonstrate that shikonin strongly inhibits cell proliferation by modulating the expression of the G1 cell cycle-related proteins, and that it induces apoptosis via the proteasome, mitochondria, and caspase cascades in SCC25 cells.