Toxoplasmosis, caused by Toxoplasma gondii (Tg), is one of the most prevalent zoonotic diseases worldwide. Currently, safe and efficient therapeutic options for this disease are still being developed, and are urgently needed. Agrimonia pilosa (A. pilosa) is a rosaceous plant, and widely distributed in Asia. The bioactive compounds in A. pilosa Ledeb include flavonoid, triterpenoids, and isocoumarin, and have been reported to show many pharmacological effects including anti-inflammatory, antioxidant, and antibacterial activities.The aim of this study was to determine Anti-protozoal efficacy of Agrimonia pilosa ethanol extract (APE) against Toxoplasma gondii (Tg). In this study, APE was examined and screened for anti-Tg activity. Seventy percent ethanol was used for herbal extraction. For anti-Tg activity screening, in vitro study with APE using tachyzoit of Tg RH strain infected HeLa cells were performed. Also, in vivo anti- Tg study using the mouse infection model was conducted. As the results, the selectivity of APE was 3.35, which was higher than Sulfadiazine selectivity (2.08). Also, we performed in vivo animal study to evaluate the anti-Tg activity of APE with mouse model. As the results, Anti-toxoplasma activity of the APE was as high as sulfadiazine. These results demonstrate that it can be successfully cured against Tg infection with APE. It could be a promising native herb treatment for the prevention of Tg infection.

The effects of antioxidant measured by DPPH radical scavenger activity, the extract of harvestry in July shown highest activity(91.8±0.01%). The all extracts of 4 parts(leaf, f1ower, root, branch) of harvestry in July were shown better radical scavenger activity than BHT(89.2±0.01%). The antibacteria activity of Agrimonia pilosa extracts was determined by using a paper disc method against bacteria, the methanol extracts(70%) showed the most active antibacteria activity against 12 kinds of harmful microorganisms. The methanol extracts(70%) showed the highest antibacterial activity against S. aureus and B. subtilis. In addition, the minimum inhibitory concentration(MIC) of methanol extracts(70%) against S. aureus, B. subtilis, C. diphtheriae, S. mutans, and S. epidermidis were revealed 0.625~10mg/mL. As a result, antibacteria activity in methanol(70%) extract of Agrimonia pilosa shown higher gram positive bacteria than gram negative bacteria.

본 연구는 짚신나물 열수 추출물의 α-glucosidase 저해 활 성을 측정하고, 분화된 근육세포에서 glucose 이용과 인슐린 신호전달에 미치는 영향을 분석하였다. 짚신나물 열수 추출 물(10 ㎎/㎖)은 α-glucosidase 활성을 67% 저해하였으며, 같은 농도의 양성대조구인 acarbose(63%)와 유사한 저해 효과를 보였다. 짚신나물 열수 추출물이 α-glucosidase에 의한 단당 류 생성을 저해함으로 식사 후 혈당이 급격히 상승하는 것을 억제하는데 효과적인 소재로 이용 가능성을 확인하였다. 또한 근육세포에서 인슐린 저항성을 유발하기 위해 지방산(1 mM, palmitic acid)를 처리하였고, glucose의 세포내 유입이 감소 되는 것을 확인하였다. 지방산 처리 세포 모델에서 짚신나 물 열수 추출물(10 ㎍/㎖)은 glucose 이용을 유의적으로 회복 시켜 주었다. Normal 상태의 배양조건에서 근육세포의 포도 당 이용능은 짚신나물 열수 추출물(100 ㎍/㎖) 처리에 의해 유의적으로 증가하였다. 근육세포 내로 glucose 유입은 운반 단백질인 Glut4를 통해 이루어지며, 이것은 인슐린이 신호전 달을 통해 조절한다. 짚신나물 열수 추출물의 세포 내 glucose 이용 증가 효과는 인슐린 신호전달 관련 분자인 Akt 유전자 와 단백질 발현을 증가시킨 것과 관련되는 것으로 추정된다. 결론적으로, 짚신나물 열수 추출물은 소화기관에서의 탄수화 물 흡수 저해와 근육세포 내 glucose 이용 증가를 통해 혈당 조절 및 당 대사 개선에 긍정적인 영향을 미치고 있음을 확인 하였다.

Weissella viridescens와 Leuconostoc carnosum은 육가공 제품 변패 미생물 중에서 중요한 미생물 로 인식되어 왔다. 용아초(Agrimonia pilosa)는 아시아에 널리 분포해 있으며, agrimonolide, agrimophol, agrimoniin, agrimol, agrimonic acid A, agrimonic acid B와 같은 다양한 플라보노이드를 함유하고 있다. 본 연구에서는 용아초의 추출방법에 따른 W. viridescens와 Leu. carnosum에 대한 생육저해효과를 탐색하였다. pH 11로 조정된 물 추출물과 50%(v/v) 에탄올 추출물은 W. viridescens 와 Leu. carnosum에 대하여 각각 43%와 24%에 해당하는 생육저해효과를 나타내었다. 그러나 사용된 추출용매와 상관 없이, 추가적인 초음파 추출은 용아초 추출물의 항균활성 증진에 큰 영향을 미치지 않았다.

선학초 메탄올 추출물을 유기용매를 이용하여 분획하고 각각의 분획별 항산화 및 항암 활성을 조사하였다. 항산화 활성으로 총페놀, 전자공여능, 아질산염 소거능을, 효소 저해 활성으로 XOase 저해 활성을 조사하였으며, 암세포인 HT-29, SNU-1, HeLa 세포에 대한 성장 저해 활성을 조사하여 기능성 식품으로의 개발 가능성을 알아보고자 하였다. 총 페놀 함량은 EtOAC와 BuOH 분획에서 39.89%, 39.56%로 가장 높았고, 전자공여능 또한 EtOAC 분획에서 92.90% (500 μg/ml), 94.47% (1000 μg/ml), BuOH 분획에서 93.77% (500 μg/ml), 92.90% (1000 μg/ml)로 90% 이상이었다. 아질산염 소거능도 1000 μg/ml의 농도에서 50% 이상이었으며, XOase 저해활성은 93.06% (EtOAC) 와 91.73% (BuOH)로 높은 활성을 보였다. 암세포 성장 저해활성을 측정한 결과 hexane 분획에서 가장 높았고 HT-29에 대해 11.63%-90.07%, SNU-1에 대해 16.84%-95.11%, HeLa에 대해 4.44%-96.40%였다. 이상의 결과에서 선학초의 EtOAc, BuOH 분획물은 항산화 활성이 있는 기능성 식품으로의 개발이 가능할 것으로 생각되며, hexane, chloroform 분획은 위암, 대장암과 자궁경부암 치료보조제로서의 개발을 위한 추가적인 연구가 필요하다고 생각된다.

Background : Agrimonia pilosa (A. pilosa) Ledebour has been registered in The Korean Herbal Pharmacopeia (KHP). In the recent study, A. Coreana showed antioxidant and anti-inflammatory effect. However, Studies of components in Agrimonia coreana (A. Coreana) Nakai was not much. So, we compared A. pilosa and A. coreana by High performance liquid chromatography (HPLC). we perfomed Thin layer chlromatography (TLC) including the anatomical characteristics by using microscope. Methods and Results : The anatomical characteristics of A. pilosa were similar to them of A. coreana. But, fascicular fivers of A. Coreana was broader than it of A. pilosa. TLC were performed to identify the Rutin and Apigenin-7-glucuronide compound. In the extract of Agrimoniae Herba, they were identified on the spot of Rf 0.2, 0.4 in Ethyl acetate - Formic acid – Water (8 : 1 : 1). The Rutin and Apigenin-7-glucuronide were analysed by HPLC/UV with Thermo Column (5 μm, 4.6 × 250 mm, C18), the column temperature at 40 ℃ and a diode-array detector (DAD) seted at 255 nm and 338 nm. The mobile phase was composed of water and acetonitrile containing 0.1% formic acid with the flow rate 1 mL/min. All compounds showed good linearity (R2>0.999) within test ranges. Agrimoniae herba was extrated by four kinds of extraction methods: MeOH, 50% MeOH, EtOH and water. The highest extraction rate occurred, when it was treated with 50% Methanol for refluxing extraction (60min). Content of Rutin was found to be 0.07±0.00 mg/g in A. pilosa and 0.02±0.00 mg/g in A. coreana. Content of Apigenin-7-glucuronide was found to be 0.12±0.00 mg/g in A. pilosa and 0.11±0.00 mg/g in A. coreana. Conclusion : The anatomical characteristics of A. pilosa were similar to them of A. coreana. Contents of Rutin and Apigenin-7-glucuronide in A. pilosa was higher than them in A. coreana slightly. But there were observed the similar patterns of Agrimonia pilosa Ledebour and Agrimonia coreana Nakai on the finger print anelysis.

Background: In this study, microwave extraction was used, which is an effective method to extract useful bioactive substances as it requires low quantities of solvent and short time periods. The aim of this study was to determine the optimal extraction conditions for Agrimonia pilosa Ledeb. Methods and Results: The independent variables were ethanol concentration, microwave power, and extraction time, each of which had five levels. The dependent variables were total polyphenol and total flavonoid content, and DPPH radical scavenging activity. To determine the optimal extraction conditions for bioactive compounds, a response surface methodology was employed. Contour maps were generated from polynomial equations. The optimal conditions were then assumed by superimposing these contour maps. Based on the resulting graph, the optimal microwave extraction conditions for Agrimonia pilosa Ledeb were determined as 42 - 48% ethanol concentration, 240 - 280W microwave power, and 13 - 20 min of extraction time. Conclusions: Ethanol concentration had a significant effect on microwave extraction, in terms of total polyphenol and total flavonoid content, as well as DPPH radical scavenging activity. Microwave power and extraction time influenced the total polyphenol content, but not the total flavonoid content or the DPPH radical scavenging activity.

본 연구에서 짚신나물(A. pilosa)의 뿌리부분에 대한 다양한 생리활성을 조사하여 기능성소재 응용가능성을 검토하였다. 짚신나물 뿌리 추출물은 멜라노마 세포에 대하여 낮은 세포독성을 나타냈다. 세포독성이 거의 없는 농도에서 짚신나물 뿌리 추출물 처리 시 항산화(ID50, 20.70 mg/L) 및 항균활성이 매우 우수하게 나타났으며 특히 그람양성세균에 대한 항균활성이 높았다. 또한, 짚신나물 뿌리 추출물 처리시 높은 tyrosinase 활성저해(ID50, 90.18 mg/L) 및 멜라닌 함량 저하를 보여주었다. 이는 짚신나물 추출물이 세포내 tyrosinase 발현을 억제시킴으로서 멜라닌 합성을 저해하는 것으로 판단된다. 이와 같은 결과로 미루어 볼 때 짚신나무 뿌리 추출물은 피부미백 소재 등 피부개선 효과를 비롯한 기능성 화장품에 활용하기 위한 매우 효과적인 재료가될 수 있다고 판단된다.

This study was performed to assess the antioxidant activities and whitening effects of Agrimonia pilosa Ledeb on melanin synthesis. The whitening effects of Agrimonia pilosa Ledeb water extracts were examined by in vitro mushroom tyrosinase assay and B16BL6 melanoma cells. We assessed inhibitory effect of Agrimonia pilosa Ledeb water extract on expression of melanogenic enzyme proteins including tyrosinase, tyrosinase-related protein 1 (TRP-1) and tyrosinase-related protein 2 (TRP-2) in B16BL6 cells. Inhibitory effect of Agrimonia pilosa Ledeb onto free radical generation was determined by measuring DPPH and hydroxyl radical scavenging activitie. Our results indicated that Agrimonia pilosa Ledeb water extract effectively inhibited free radical generation. In DPPH and hydroxy radical scavenging activity, Agrimonia pilosa Ledeb water extract had a potent anti-oxidant activity in a dose-dependent manner. They significantly inhibited tyrosinase activity in vitro and in B16BL6 melanoma cells. Also, Agrimonia pilosa Ledeb suppressed the expression of tyrosinase in B16BL6 melanoma cells. These results show that Agrimonia pilosa Ledeb inhibited melanin production on the melanogenesis. The underlying mechanism of Agrimonia pilosa Ledeb on whitening activity may be due to the inhibition of tyrosinase activity. We suggest that Agrimonia pilosa Ledeb may be useful as new natural active ingredients for antioxidant and whitening cosmetics.

Complementary and alternative medicines are considered as a promising research field to develop new therapies for various allergic diseases. In this study, we investigated the anti-allergic effect of Agrimonia pilosa Ledeb (AP) by using passive cutaneous anaphylaxis in mice and its mechanism of action in mast cells. The extract of AP reversibly inhibited degranulation in RBL-2H3 cells and bone marrow-derived mast cells (BMMCs). AP also suppressed the passive cutaneous anaphylaxis inducing by IgE and antigen (Ag) in a dose-dependent manner. In the study to find its mechanism of action, AP inhibited the phosphorylation of Syk kinase, a pivotal protein which is regulated by Src-family kinase for activation of mast cells. In addition, AP also suppressed activation of Akt and Erk1/2 that are critical for the production of cytokines in mast cells. The results strongly suggest that AP exerts anti-allergic activity in vitro and in vivo through the inhibition of activation of Syk in mast cells.

Changes in the contents of total phenolic compounds in as Agrimonia pilosa well as their antioxidant capacity according to the havest time and positions were examined. The contents of the total phenolic compounds were determined by extraction with MeOH. Among havest times harvestry in July showed highest contents of the total phenolic compounds and harvestry in May showed lowest contents of the phenolic compounds. Among the 4 positions (root, branch, leaf, flower) of Agrimonia pilosa the root contained highest contents of the phenolic compounds. The antioxidant capacities of Agrimonia pilosa were increased roughly with increasing level of contents of phenolic compounds according to positions.

The antioxidant activities of Agrimony (Agrimonia pilosa L.) and Chinese lizardtail (Saururus chinensis Baill) according to extraction methods were measured. SOD-like activity showed greater antioxidant effects with ethanol solvent than those with water. Ethanol extracts of Agrimony leaves showed the highest SOD-like activity of 94.4%. SOD-like activity differed according to the extraction solvents. The contents of polyphenolic compounds were higher in water extracts than those in ethanol extracts. The contents were 161.4 mg for Agrimony roots, 100.2 mg for Agrimony leaves, and 79.1 mg for Agrimony stalks in order. EDA in Agrimony leaves that were highest among medicinal plants were 83.4% in the water extract and 81.7% in the ethanol extract. The anticancer effects of the extracts by water and ethanol from Agrimony and Chinese lizardtail were experimented. The growth of stomach cancer cells, SNU-719 was inhibited 94.5% by the hexane fractions of Agrimony and also the growth of liver cancer cells, Hep3B was inhibited 83.2% by the hexane fractions of Agrimony, while the growth of normal cell, DC2.4 was not affected.