동백 유래 추출물의 구강보건효능 및 생체안전성 분석

Abstract

This study evaluates the antioxidant and anti-inflammatory effects of camellia leaf and flower extracts. The safety evaluation was conducted through oral administration to rats, and the potentiality of the extracts as oral health products was analyzed. The antioxidant activity of 70% ethanol extracts of camellia leaves and flowers was analyzed by four methods. As a result of confirming the ABTS radical inhibitory effect of the camellia leaf and flower extracts, the inhibitory effects of 96.66 ± 0.84% and 97.17% were confirmed in the camellia leaf and flower extracts at a concentration of 1 mg/㎖ respectively. In addition, as a result of analyzing DPPH radical scavenging activity, it was confirmed that the inhibitory effect was 93.71% in the camellia leaf extract and 63.50% in the camellia flower extract at a concentration of 1 mg/㎖. As a result of confirming the SOD-like activity, the activity was 44.27% in the camellia leaf sample at 5 mg/㎖ concentration and 23.69% in the camellia flower sample at 5 mg/㎖ concentration. As a result of analyzing the total polyphenol content and the total flavonoid content in order to determine how much of the above antioxidant-related compounds are contained in the camellia extract, the total polyphenol content in the camellia flower sample at a concentration of 1 mg/㎖ is tannic acid standard. As a result of measuring as a curve, the flavonoid content of 139.96 ± 0.75 μg/㎖ and naringin as a standard curve was measured as 53.70 ± 2.57 μg/㎖. In addition, mouse macrophage RAW 264.7 cells were used to determine the anti-inflammatory activity of camellia extract. As a result of treatment with 1, 5, 10, 20, 50 and 100 μg/㎖ of camellia leaf extract on cells induced by LPS, NO production up to 3.37, 2.71, 2.66, 2.54, 2.10 and 1.41μM, respectively, by concentration In the camellia flower extract, it was confirmed that NO generation was suppressed to 4.63, 4.25, 3.77, 3.49, 2.03, and 1.51μM, respectively,

In addition, in this experiment, in order to verify the biosafety of the camellia extract, rats were used as experimental animals, and the non-toxicity of the extract was confirmed by oral administration. No dead animals were observed in all test groups during the experiment. Also, no specific general symptoms were observed in all test animals during the experiment. As a result of body weight measurement, in one case of the female 1,000 mg/kg administration group (G3-31), the body weight on the 14th day after administration of the test substance decreased slightly compared to the 7th day after administration. In other test animals, normal weight gain was observed. As a result of statistical analysis, no significant difference in weight was observed between the groups in both male and female test groups. As a result of autopsy at the end of the experiment, no peculiar macroscopic findings were observed.

The experimental results, verified that natural products derived from camellia leaves and flowers are valuable as natural oral care materials and can be safely administered in vivo. Camellia extract is expected to be available as a plant-derived natural material for various oral health products, which will be increasingly used in future. Therefore, camellia, which is a special plant in the Honam Korea region , will provide added value to the natural products industry.|실험 배경 및 목적: 동백나무(Camellia japonica L.)의 잎과 꽃 추출물의 항산화 및 항염증 활성은 많이 알려져 있다. 또한, 병원성 미생물에 대한 항균 활성을 가지고 있다. 특히, 구강 내에 서식하는 구취균과 충치균에 대한 항균활성을 가지고 있어 본 연구에서는 동백나무 잎과 꽃 추출물의 항산화와 항염증의 효과를 평가하고 랫드 경구투여를 통해 안전성 평가를 진행하였으며 구강보건용품으로 활용 가능성을 분석하였다.

방법: 본 실험에서 동백나무 잎과 꽃에 항산화합물의 함량은 Folin & Denis방법으로 분석하였고 총 페놀(Polyphenol) 과 플라보노이드(Flavonoid)의 함량을 μg/㎖로 표기 하였다. 항산화능은 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 2,2-diphenyl-1-picrylhydrazyl(DPPH), superoxide dismutase (SOD)활성을 평가하였으며, RAW 264.7 대식세포에 Lipopolysaccharides (LPS) 처리한 후 염증 유도인자인 nitric oxide (NO)의 저해활성을 평가하였다. 추출물의 인체안정성을 확인하기 위해 Sprague-Dawley (SD) rat를 이용하여 경구독성시험을 수행하였다. 추출물을 적용한 세치제 및 구강청결제를 제조하여 물성을 평가하였다.

결과: 본 실험에서 동백 잎과 꽃 추출물에 존재하는 항산화 화합물인 폴리페놀화합물은 tannic acid을 표준물질로 측정하였으며 동백 잎과 꽃 추출물에서 각각 186.42 ± 2.17 μg/㎖ 및 139.96 ± 0.75 μg/㎖을 함유하는 것으로 나타났다. Naringin을 표준물질로 측정한 폴라보노이드의 경우 동백 잎과 꽃의 추출물은 각각 116.06 ± 1.29 μg/㎖ 및 53.70 ± 2.57 μg/㎖로 분석되었다. ABTS radica 소거능은 1 mg/㎖의 농도에서 동백잎은 96.66 ± 0.84%, 동백꽃은 96.43 ± 0.04%의 소거율을 나타내었다. DPPH radical 소거능의 경우 1 mg/㎖의 농도에서 동백잎은 93.71 ± 4.73%, 동백꽃은 63.50 ± 1.42%의 억제 효과를 확인하였다. SOD의 소거능을 측정한 결과 5 mg/㎖의 농도에서 동백잎은 44.27 ± 0.90%, 동백꽃은 23.70 ± 2.51%의 활성도를 확인하였다. 대체적으로 동백꽃보다는 동백잎에서 높은 활성을 나타냈었다. RAW 264.7 세포에서 LPS를 처리한 후 NO생성량을 측정한 결과 대조군은 5.34μM로 분석된 반면, 동백잎 시료를 1, 5, 10, 20, 50 및 100 μg/㎖로 처리한 결과 농도별로 각각 3.37, 2.71, 2.66, 2.54, 2.10 및 1.41μM까지 NO 생성량이 감소하였고, 동백꽃 추출물에서는 농도별로 각각 4.63, 4.25, 3.77, 3.49, 2.03 및 1.51μM까지 NO 생성량이 감소하였다. 추출물을 경구 투여한 rat에서 실험기간 동안 모든 시험군에서 사망동물이 관찰되지 않았다. 또한 실험기간 동안 모든 시험동물에서 특이한 일반증상이 관찰되지 않았다. 이상의 실험결과를 토대로 동백나무 잎과 꽃 유래 천연물이 천연 구강케어소재로서 충분한 가치가 있고, 생체 내에서도 안전하다는 것을 검증하였다. 이와 같은 실험결과를 토대로 향후 다양한 구강보건용품 개발에 필요한 식물유래 천연소재로서 동백나무 유래 천연물의 활용가치가 높아질 것으로 사료되며, 천연물산업에서 호남지역 특산식물인 동백나무의 산업적 부가가치가 향상될 것으로 사료된다