Hyaluronidase Inhibitory Activity Research Articles

번행초(Tetragonia tetragonioides)로부터 추출한 Phenolic Compounds의 미용 식품 활성

본 연구는 물과 ethanol을 추출용매로 사용한 번행초(Tetragonia tetragonioides) 추출물이 미용 식품 소재로 사용 가능한 기능성을 증명하고자 하였다. Phenolic compounds는 물 추출물에서 3.29 mg/g, 50% ethanol 추출물에서 4.14 mg/g의 결과를 각각 나타내었다. 전자공여능 측정 결과 200 μg/mL phenolic compounds의 농도에서 물 추출물은 98.45%, 50% ethanol 추출물은 91.20%의 활성을 나타내었다. ABTS radical 활성 측정 결과 100 μg/mL phenolic compounds의 농도에서 물 추출물은 97.28%, 50% ethanol 추출물은 97.03%의 활성을 나타내었다. Antioxidant protection factor(PF) 측정 결과 200 μg/mL phenolic compounds의 농도에서 물과 ethanol 추출물 모두 1.77 PF의 같은 결과를 나타내었다. Thiobarbituric acid reactive substance(TBARS) 측정 결과 100 μg/mL phenolic compounds의 농도에서 물과 ethanol 추출물은 94.77%와 95.64%의 높은 활성을 나타내었다. 미백과 관련 있는 tyrosinase 저해 활성을 측정한 결과 200 μg/mL phenolic compounds의 농도에서 ethanol 추출물은 34.96%의 억제력을 나타내었다. 주름 개선 효과를 측정하는 elastase 저해 활성과 collagenase 저해 활성을 측정한 결과 200 μg/mL phenolic compounds의 농도에서 ethanol 추출물만 78.90%와 61.3%의 높은 저해 활성을 나타내었다. 수렴 효과를 나타내는 astringent effect에서 200 μg/mL phenolic compounds 농도의 ethanol 추출물은 7.82%의 효과를 나타내었다. Hyaluronidase 저해 효과 측정 결과 물 추출물 200 μg/mL phenolic compounds의 농도에서 81.04%의 염증 억제 효과를 나타내었다. 이러한 결과들로 미루어 보았을 때 번행초 추출물은 항산화 작용과 미용 식품으로서의 기능성을 기대할 수 있다.

Hyaluronidase Inhibitory Activity of Pentacylic Triterpenoids from Prismatomeris tetrandra (Roxb.) K. Schum: Isolation, Synthesis and QSAR Study.

The mammalian hyaluronidase degrades hyaluronic acid by the cleavage of the β-1,4-glycosidic bond furnishing a tetrasaccharide molecule as the main product which is a highly angiogenic and potent inducer of inflammatory cytokines. Ursolic acid 1, isolated from Prismatomeris tetrandra, was identified as having the potential to develop inhibitors of hyaluronidase. A series of ursolic acid analogues were either synthesized via structure modification of ursolic acid 1 or commercially obtained. The evaluation of the inhibitory activity of these compounds on the hyaluronidase enzyme was conducted. Several structural, topological and quantum chemical descriptors for these compounds were calculated using semi empirical quantum chemical methods. A quantitative structure activity relationship study (QSAR) was performed to correlate these descriptors with the hyaluronidase inhibitory activity. The statistical characteristics provided by the best multi linear model (BML) (R2 = 0.9717, R2cv = 0.9506) indicated satisfactory stability and predictive ability of the developed model. The in silico molecular docking study which was used to determine the binding interactions revealed that the ursolic acid analog 22 had a strong affinity towards human hyaluronidase.

Antioxidant and hyaluronidase inhibitory activities of diverse phenolics in Phyllanthus emblica

Fifty-eight phenolic compounds isolated from Phyllanthus emblica were screened and compared for their in vitro and in vivo antioxidant properties, as well as hyaluronidase (HAase) inhibitory activities. Among them, 20 compounds showed to be promising antioxidants due to the stronger scavenging activity in both DPPH radical and Danio rerio reactive oxygen species assays, while nine compounds were potential HAase inhibitors with 100-fold stronger activities than that of the positive control, DSCG. The structure activity relationship was discussed.

Anti-hyaluronidase Activity in Vitro and Amelioration of Mouse Experimental Dermatitis by Tomato Saponin, Esculeoside A.

The increasing incidence of atopic dermatitis during recent decades has prompted the development of safe and effective agents for prevention of atopic diseases. Esculeoside A, a glycoside of spirosolane type, is identified as a major component in ripe tomato fruits. The present study investigated the effects of esculeoside A and its aglycon esculeogenin A on hyaluronidase activity in vitro and antiallergy in experimental dermatitis mice. Esculeogenin A/esculeoside A (esculeogenin A equivalent) with an IC50 of about 2 μM/9 μM dose-dependently inhibited hyaluronidase activity measured by a modified Morgan-Elson method. Oral treatment with esculeoside A 10 mg/kg of experimental dermatitis mice for 4 weeks significantly decreased the skin clinical score to 2.5 without any detectable side effects compared with 6.75 of the control. The scratching frequency of esculeoside A 100 mg/kg application was decreased significantly as 107.5 times compared with 296.67 times of the control. Thus, the present study showed that esculeoside A/esculeogenin A significantly blocks hyaluronidase activity in vitro and that esculeoside A ameliorates mouse experimental dermatitis.

Biological Activities of Extracts fromCornus kousaFruit

The extracted phenolic compounds from Cornus kousa fruit for biological activities as functional resources were examined. The phenolic compounds which were extracted with water and 40% ethanol from Cornus kousa fruit were 7.04±0.27 and 4.47±0.18 mg/g, respectively. The 1,1-diphenyl-2-picrylhydrazyl free radical scavenging activity of water and ethanol extracts were 84% and 86% at 50 µg/mL phenolics, respectively. The 2,2'- Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid radical decolorization activity of water and ethanol extracts were 84 and 95% at 100 µg/ mL phenolics, respectively. Antioxidant protection factor in water and ethanol extracts at 50 µg/mL phenolics were 1.93 and 1.82 PF, respectively. Thiobarbituric acid reactive substance were 69% in water extracts and 89% in ethanol extracts at 150 µg/mL phenolics. The inhibition activity on xanthine oxidase in water and ethanol extracts was 34 and 60%, respectively. The inhibition activity on α-glucosidase was 29% in water extracts and 87% in ethanol extracts. The tyrosinase inhibitory activity was 19% in ethanol extracts. The collagenase inhibition activity of anti-wrinkle effect showed an excellent wrinkle improvement effect as 53% in water extracts and 77% in ethanol extracts at 200 µg/mL phenolics. The hyaluronidase inhibition activity as antiinflammation effect of water extracts was confirmed to 34% of inhibition at 200 µg/mL phenolic. The results can be expected extracts from Cornus kousa fruit to use as functional resource for antioxidant, antigout, inhibitor of carbohydrate degradation, antiwrinkle activity and antiinflammation activity.

Phenolic constituents of the Bangladeshi medicinal plant Pothos scandens and their anti-estrogenic, hyaluronidase inhibition, and histamine release inhibitory activities

Extracts from the stem and roots of the Bangladeshi medicinal plant Pothos scandens L. (Araceae) were isolated, and three hemiterpene glucoside aromatic esters, pothobanosides A (1), B (2), and C (3), and a phenylisobutanoid, pothobanol (4), along with 14 known compounds, were characterized. The isolates were tested for their estrogenic/anti-estrogenic activity using the estrogen-responsive human breast cancer cell lines MCF-7 and T47D, and syringoyl derivatives (2, 3, and canthoside B) showed strong inhibitory activity against both cell lines. Their less oxygenated analogs (1, and markhamioside F) were almost inactive. The isolates were also evaluated for hyaluronidase and histamine release inhibitory activities, and pothobanoside A (1) showed significant hyaluronidase inhibitory activity among the isolated compounds, which was similar to that of the positive control rosmarinic acid. Because hyaluronidase produces an angiogenic response that has been implicated in tumor invasiveness and metastasis, 1 could be valuable as an anti-tumor compound with a different mechanism of action from related compounds (2, 3). Pothobanoside C (3) and pothobanol (4) were also found to inhibit histamine release to a similar degree to the positive control epigallocatechin 3-O-(3″-O-methyl)-gallate. The histamine release inhibitory potency of these isolates may support the traditional uses of this plant in folk medicine.

Isolation and analysis of polysaccharide showing high hyaluronidase inhibitory activity in Nostochopsis lobatus MAC0804NAN

An active substance with high hyaluronidase inhibitory effect was isolated from the edible cyanobacterium Nostochopsis lobatus MAC0804NAN strain and characterized. The active component in the hot water extract was purified by anion exchange and gel filtration chromatography and was found to be a polysaccharide. The IC(50) against hyaluronidase of the purified polysaccharide was 7.18 μg/ml whose inhibitory activity is 14.5 times stronger than that of disodium cromoglycate (DSCG), an anti-allergy medication. The carbohydrate composition which was analyzed by GC-MS and NMR was found to be composed mainly of glucose, glucuronic acid, fucose, 2-O-methylfucose, mannose, galactose and xylose.

In vitro inhibition of hyaluronidase by sodium copper chlorophyllin complex and chlorophyllin analogs.

BackgroundInhibitors of hyaluronidase are potent agents that maintain hyaluronic acid homeostasis and may serve as anti-aging, anti-inflammatory, and anti-microbial agents. Sodium copper chlorophyllin complex is being used therapeutically as a component in anti-aging cosmeceuticals, and has been shown to have anti-hyaluronidase activity. In this study we evaluated various commercial lots of sodium copper chlorophyllin complex to identify the primary small molecule constituents, and to test various sodium copper chlorophyllin complexes and their small molecule analog compounds for hyaluronidase inhibitory activity in vitro. Ascorbate analogs were tested in combination with copper chlorophyllin complexes for potential additive or synergistic activity.Materials and methodsFor hyaluronidase activity assays, dilutions of test materials were evaluated for hydrolytic activity of hyaluronidase by precipitation of non-digested hyaluronate by measuring related turbidity at 595 nm. High-performance liquid chromatography and mass spectroscopy was used to analyze and identify the primary small molecule constituents in various old and new commercial lots of sodium copper chlorophyllin complex.ResultsThe most active small molecule component of sodium copper chlorophyllin complex was disodium copper isochlorin e4, followed by oxidized disodium copper isochlorin e4. Sodium copper chlorophyllin complex and copper isochlorin e4 disodium salt had hyaluronidase inhibitory activity down to 10 µg/mL. The oxidized form of copper isochlorin e4 disodium salt had substantial hyaluronidase inhibitory activity at 100 µg/mL but not at 10 µg/mL. Ascorbate derivatives did not enhance the hyaluronidase inhibitory activity of sodium copper chlorophyllin. Copper isochlorin e4 analogs were always the dominant components of the small molecule content of the commercial lots tested; oxidized copper isochlorin e4 was found in increased concentrations in older compared to newer lots tested.ConclusionThese results support the concept of using the hyaluronidase inhibitory activity of sodium copper chlorophyllin complex to increase the hyaluronic acid level of the dermal extracellular matrix for the improvement of the appearance of aging facial skin.

Enzymatic synthesis of hyaluronan hybrid urinary trypsin inhibitor

Human urinary trypsin inhibitor is a proteoglycan that has a single low-sulfated chondroitin 4-sulfate chain at the seryl residue in position 10 of the core protein as a glycosaminoglycan moiety, and is used as an anti-inflammatory medicine based on the protease inhibitory activity of the core protein. However, the functions of the glycosaminoglycan moiety have not yet been elucidated in detail. In the present study, the glycosaminoglycan chains of a native urinary trypsin inhibitor were remodeled to hyaluronan chains, with no changes to the core protein, using transglycosylation as a reverse reaction of the hydrolysis of bovine testicular hyaluronidase, and the properties of the hybrid urinary trypsin inhibitor were then analyzed. The trypsin inhibitory activitiy of the hyaluronan hybrid urinary trypsin inhibitor was similar to that of the native type; however, its inhibitory effect on the hydrolysis of hyaluronidase were not as strong as that of the native type. This result demonstrated that the native urinary trypsin inhibitor possessed hyaluronidase inhibitory activity on its chondroitin sulfate chain. The hyaluronan hybrid urinary trypsin inhibitors obtained affinity to a hyaluronan-binding protein not exhibited by the native type. The interactions between the hyaluronan hybrid urinary trypsin inhibitors and phosphatidylcholine (abundant in the outer layer of plasma membrane) were stronger than that of the native type. Hyaluronan hybrid urinary trypsin inhibitors may be useful for investigating the functions of the glycosaminoglycan chains of urinary trypsin inhibitors and hyaluronan, and our hybrid synthesizing method may be used widely in research for future medical applications.

ISOLATION AND CHARACTERIZATION OF A NOVEL HYALURONIDASE INHIBITOR FROM A MARINE ACTINOMYCETES STRAIN

A novel hyaluronidase inhibitor (HI) was isolated from the culture extract of a marine- derived actinomycete strain. This strain MB-PO13 was isolated from ascidian (Molgula manhattensis) in Tokyo Bay. Out of about 1,000 isolates from various marine organisms, strain MB-PO13 had the strongest inhibitory activity and was selected for further study. The strain showed abundant-to-moderate growth on most media, forming a grayish mycelium. On the basis of the taxonomical characteristics, the strain was classified as belonging to the genus of Streptomyces and was named as Streptomyces sp. strain MB-PO13. The structure of HI was elucidated by interpretation of NMR data. HI displayed about 25-fold potent hyaluronidase inhibitory activity against hyaluronidase than glycyrrhizin. Keywords: marine actinomycetes; Streptomyces; hyaluronidase inhibitor

Production and characterisation of hyaluronidase and elastase inhibitory protein hydrolysates from Venus clam

The hydrolysates of fresh and boiled Venus clams with five different proteases for the production of low-molecular protein hydrolysates were optimised by response surface methodology. Alcalase hydrolysates exhibited the strongest hyaluronidase inhibitory activity. The optimum hydrolysis conditions of fresh and boiled clams were< enzyme-to-substrate ratio (E/S), 2.15%; time, 150 min; water-to-substrate ratio (W/S), 83.84 mL g− 1 for fresh clam, and E/S, 2.02%; time, 4.11 h; W/S, 69.74 mL g− 1 for boiled clam. The fresh and boiled clam protein hydrolysates were fractionated by S-200 HR size-exclusion chromatography, which resulted in one (FH1) and two (BH1 and BH2) fractions, respectively. BH1 exhibited the highest hyaluronidase and elastase inhibitory activities with specific activities of 141.15 and 81.36% mL mg− 1, respectively. Therefore, the boiled Venus clam hydrolysate might be developed as a cosmeceutical agent because of its strong hyaluronidase and elastase inhibitory activities.

Antioxidative and anti-inflammatory activities of Citrus unshiu peel extracts using a combined process of subcritical water extraction and acid hydrolysis

Antioxidant and in vitro anti-inflammatory activities of citrus (Citrus unshiu) peel extracts and their acid hydrolysates were evaluated. Citrus peel extracts were extracted by subcritical water, hot water, and ethanol. Subcritical water extraction led to extract more phenolic compounds than hot water or ethanol extraction. Acid hydrolysis more than doubled the extracts’ total phenolics and flavonoids. Acid hydrolysates showed greater DPPH-radical scavenging activities and antioxidant activities, as assayed by β-carotene bleaching and ferric thiocyanate, than the initial extracts. Anti-inflammatory activities of citrus peel extracts and hydrolysates, determined by the inhibition of hyaluronidase activity, showed that the inhibition activities of hot water and ethanol extracts increased from 2.1 and 1.8% to 37.0 and 18.5%, respectively, upon acid hydrolysis; however, the anti-inflammatory activity of the subcritical water extract was not improved. These results indicated that acid hydrolysis of citrus peel extracts regardless of their extraction methods improved the antioxidative and anti-inflammatory activities.

Monoterpene glycosides, phenylpropanoids, and acacetin glycosides from Dracocephalum foetidum

Chemical investigation of the acetone extract from the aerial parts of the Mongolian medicinal plant Dracocephalum foetidum resulted in the isolation of three limonene glycosides, a caffeic acid trimer, four rosmarinic acid glucosides, and five acacetin acyl glycosides, together with 13 known natural products. The chemical structures of all of the compounds were determined by spectroscopic analyses. Among these compounds three showed hyaluronidase inhibitory activity. In addition, one other compound showed stronger 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity than the positive control Trolox, whereas three other compounds demonstrated a similar activity to that of Trolox.

미세분쇄에 의한 감국(Chrysanthemum incidicum Linne) 추출물의 생리활성

감국 phenolic compound의 추출을 위한 최적 추출조건은 70% ethanol로 6시간 추출이 최적이었다. 그때 추출수율은 물 추출물이 7.12±1.61 mg/g, ethanol 추출물이 7.51±2.14 mg/g이었고 초미세분쇄 했을 경우 물 추출물이 8.63±1.15 mg/g, ethanol 추출물이 9.33±1.35 mg/g이었다. 감국 추출물의 항산화력 측정에서 DPPH는 일반분쇄 추출물이 83.52%, 초미세분쇄 추출물이 92.37%이었다. ABTS radical cation decolorization의 경우 일반분쇄, 미세분쇄 및 초미세분쇄 추출물 모두 90% 이상이었다. Antixodant protection factor(PF)의 경우 초미세분쇄 한 감국의 물과 에탄올 추출물에서 1.82 PF와 2.16 PF의 높은 항산화력을 나타내었다. TBARS도 일반분쇄 및 미세분쇄에 비해 초미세분쇄 한 감국 추출물에서 더 낮은 TBARS값을 나타내었다. 감국 추출물의 xanthin oxidase 저해효과는 초미세분쇄물 추출물이 67.53%, ethanol 추출물이 83.45%였다. Xanthin oxidase 저해는 물 추출물보다는 ethanol 추출물이, 초미세분쇄 추출물이 일반 추출물에 비해 효소억제효과가 더 우수한 결과를 나타내었다. Angiotensin converting enzyme 저해효과는 초미세분쇄 추출물의 경우 물 추출물에서 24% 이상의 억제효과를 나타내었고, ethanol 추출물은 34%의 억제효과를 나타내었다. 감국 추출물의 elastase 억제효과는 미세분쇄 추출물의 저해율 20.34%에 비하여 초미세분쇄 추출물의 억제율이 25.56%로 더 우수한 기능성을 나타내었다. Hyaluronidase의 경우 감국 물 추출물에서는 억제효과가 관찰되지 않았으며, ethanol 추출물에서만 35% 정도의 염증억제효과를 보여주었다. 감국 추출물에 의해 대식세포에서 iNOS와 COX-2 단백질 발현억제 효과를 Western blot analysis로 측정한 결과 감국 에탄올 추출물에 의해 iNOS와 COX-2 단백질 발현이 100 μg/mL의 농도에서 각각 40%와 15%의 발현억제를 나타내었고, 농도 의존적으로 억제됨을 확인하였다. 본 연구결과 초미세분쇄에 의한 감국 추출물은 일반분쇄 추출물에 비해 고혈압과 통풍억제 효과가 확연히 높아짐을 알 수 있었다.

Hyaluromycin, a New Hyaluronidase Inhibitor of Polyketide Origin from Marine Streptomyces sp.

Hyaluromycin (1), a new member of the rubromycin family of antibiotics, was isolated from the culture extract of a marine-derived Streptomyces sp. as a HAase inhibitor on the basis of HAase activity screening. The structure of 1 was elucidated through the interpretation of NMR data for the compound and its 3″-O-methyl derivative in combination with an incorporation experiment with [1,2-13C2]acetate. The compound’s absolute configuration was determined by the comparison of its circular dichroism (CD) spectrum with those of other rubromycins. Hyaluromycin (1) consists of a γ-rubromycin core structure possessing a 2-amino-3-hydroxycyclopent-2-enone (C5N) unit as an amide substituent of the carboxyl function; both structural units have been reported only from actinomycetes. Hyaluromycin (1) displayed approximately 25-fold more potent hyaluronidase inhibitory activity against hyaluronidase than did glycyrrhizin, a known inhibitor of plant origin.

Effect of balanced low pressure drying of curcuma longa leaf on skin immune activation activities.

The effect of balanced low pressure drying pretreatment associated with ultrasonication extraction (BU) on the enhancement of skin immune modulatory activities of Curcuma longa leaf was studied by comparing with conventional hot air drying (HE), freeze drying (FE) and balanced low pressure drying (BE) pretreatment processes. In considering skin immune activation activities such as the inhibition of hyaluronidase activity, the BU extract showed ca. 10% higher than those of HE, and even higher than that of the FE extract. Nitric oxide production from macrophage of the BU extract in adding 1.0 mg/mL was increased up to 16.5 μM. When measuring inhibition of IL-6 and TNF-a production from the human T lymphocytes (T cell), the BU extract also showed 53% and 78% of inhibition effect, respectively. It is found that the BU extract could effectively suppress the expression levels of skin inflammation related genes such as Cox-2 and iNOS, down to 80% and 85% compared to the control, respectively. Balanced low pressure drying process was especially active on dehydration of the leaves with minimizing the destruction and making easier elution of the bioactive substances, which resulted in higher extraction yield and better biological activities.

유산균 발효를 통한 황금 추출물의 항염증 효과 증진

본 연구에서는 소염, 항염 작용이 뛰어난 작물로 알려진 황금을 유산균인 Lactobacillus rhamnosus를 이용해 발효를 통한 황금의 추출물에 대한 추출공정별 항염증 효과 증진을 측정하였다. 추출 공정으로는 일반열수추출(WE), 일반 70% 에탄올 추출(EE), L. rhamnosus 발효 열수 추출(FWE), L. rhamnosus 발효 70% 에탄올 추출(FEE)로 나누어 진행하였다. 각각의 추출 공정별 수율 측정결과 발효 70% 에탄올 추출이 25.9%로 최대 수율을 얻었으며, 발효 열수 추출, 일반 70% 에탄올 추출, 일반 열수 추출 순으로 높은 수율을 얻었다. 추출 공정별 세포독성 측정에서 발효 황금 에탄올 추출물의 농도가 1.0 mg/mL일때 9.8%로 가장 낮은 세포독성을 나타냈으며, 일반 에탄올 추출의 경우 22.6%로 가장 높은 세포 독성을 가졌다. Hyaluronidase 저해 효과는 발효 황금 에탄올 추출물의 농도가 1.0 mg/mL에서 최대 46.8%의 hyaluronidase 저해 활성을 보였으며, NO 생성량 및 <TEX>$PGE_2$</TEX> 생성량 측정도 1.0 mg/mL 농도의 발효 황금 에탄올 추출물이 각각 36%의 최대 NO 생성량 감소 및 810 pg/mL의 낮은 <TEX>$PGE_2$</TEX> 값을 나타냈다. 따라서 황금의 항염증 관련 효과를 위한 최적 추출 공정으로는 발효를 통한 황금의 70% 에탄올 추출 수율이 가장 높았으며, 이에 따라 항염증 관련 생리활성 물질이 보다 더 많이 용출됨을 확인할 수 있다. This study was performed to investigate the anti-inflammatory activities of fermented Scutellaria baicalensis extracts using Lactobacillus rhamnosus. The extracts were WE (water extract at <TEX>$100^{\circ}C$</TEX> for 24 h), EE (70% ethanol extract at <TEX>$60^{\circ}C$</TEX> for 24 h), FWE (fermented and water extract at <TEX>$60^{\circ}C$</TEX> for 24 h), FEE (fermented and 70% ethanol extract at <TEX>$60^{\circ}C$</TEX> for 24 h). The cytotoxicity of the extracts was in the range of 11.2 ~ 15.6 % at 1.0 mg/mL concentratioin. The FEE showed the lowest activity at 1.0 mg/mL concentratioin. Compared to the WE, hyaluronidase inhibitory activity contents in the FEE increased to 9.2% at 1.0 mg/mL concentratioin. Nitric oxide production of WE, EE, FWE and FEE at 1.0 mg/mL concentration was mesured as 7.6, 7.9, 6.9, 6.4 <TEX>${\mu}M$</TEX>, respectively. <TEX>$PGE_2$</TEX> secretion of the human fibroblast of the FEE were lower than 810 pg/mL. Our results suggested that the extracts from fermentation process after 70% ethanol extraction had relatively high anti-inflammatory activities and that the Scutellaria baicalensis could be more extracted in FEE than others.

Improved cosmetic activity by optimizing the Lithospermum erythrorhizon extraction process

This study was conducted to expand the use of Lithospermum erythrorhizon, which is a good source of natural dye, in skin whitening and immune activation cosmetics. The goal was to provide cosmeceutical data about the extraction yield and shikonin contents of this plant by optimizing the ultrasonic extraction and high pressure extraction conditions. Under optimal extraction conditions, which consisted of 500MPa for 60min and 120kHz for 90min, 27.49 and 3.19% (w/w) of the highest extraction yield and shikonin contents were obtained, compared to 16.32 and 1.81% from a conventional ethanol extract (EE) control. Hyaluronidase inhibition activity was measured as 44.24% after adding 1.0mg/ml of ethanol extract, but it was as high as 64.19% when using extract produced by ultrasonication with high pressure extraction (UE+HPE). The MMP-1 expression levels from skin fibroblast cells (CCD-986sk) treated with or without UV irradiation were also lowered by as much as 110.6% after adding 1.0mg/ml of the UE+HPE extract, relative to 126.9% from the EE. After UVA exposure, prostaglandin E2 production from RAW 264.7 was also lower, at 110.6%, which also indicates that the extract from the UE+HPE process enhanced skin immune activation activities. For the skin whitening activity, tyrosinase inhibitory activity was observed at 67.15% in the HPE+UE extract, which was ca. 20% higher than that of the EE extract (57.48%). To reduce melanin production in Clone M-3 cells, 79.5% of the melanin production was estimated after adding 1.0mg/ml of the UE+HPE extract compared to that of the control (no treatment), which was similar to the 77.4% result found in an ascorbic acid positive control. The highest shikonin secretion was conclusively obtained under the optimal conditions and resulted in a significant improvement of the cosmetic activities of L. erythrorhizon extracts.

Effects of Payena dasyphylla (Miq.) on hyaluronidase enzyme activity and metalloproteinases protein expressions in interleukin-1β stimulated human chondrocytes cells.

BackgroundHyaluronidases have been found as the target enzymes in the development of osteoarthritis (OA) disease. While there is still no curative treatment for this disease, recent studies on the treatment of OA were focused on the effectiveness of natural products which are expected to improve the symptoms with minimal side effects. The aim of this study was to screen selected Malaysian plants on their anti-hyaluronidase activity as well as to evaluate the active plant and its derived fractions on its potential anti-arthritic and antioxidant activities.MethodsA total of 20 methanolic crude extracts (bark and leaf) from ten different plants were screened using a colorimetric hyaluronidase enzymatic assay. The active plant extract (Payena dasyphylla) was then studied for its hyaluronidase inhibitory activity in the interleukin-1β (IL-1β) stimulated human chondrocytes cell line (NHAC-kn) using zymography method. The Payena dasyphylla methanolic bark extract was then fractionated into several fractions in where the ethyl acetate (EA) fraction was evaluated for its inhibitory effects on the HYAL1 and HYAL2 gene expressions using reverse transcription-polymerase chain reaction (RT-PCR) technique. While the MMP-3 and MMP-13 protein expressions were evaluated using western blot method. The phenolic and flavonoid contents of the three fractions as well as the antioxidant property of the EA fraction were also evaluated.ResultsBark extract of Payena dasyphylla (100 μg/ml) showed the highest inhibitory activity against bovine testicular hyaluronidase with 91.63%. The plant extract also inhibited hyaluronidase expression in the cultured human chondrocyte cells in response to IL-1β (100 ng/ml). Similarly, treatment with Payena dasyphylla ethyl acetate (EA) fraction (100 μg/ml) inhibited the HYAL1 and HYAL2 mRNA gene expressions as well as MMP-3 and MMP-13 protein expression in a dose dependent manner. Payena dasyphylla EA fraction has demonstrated the highest amount of phenolic and flavonoid content with 168.62 ± 10.93 mg GAE/g and 95.96 ± 2.96 mg RE/g respectively as compared to water and hexane fractions. In addition, the Payena dasyphylla EA fraction showed strong antioxidant activity with IC50 value of 11.64 ± 1.69 μg/mL.ConclusionThese findings have shown that Payena dasyphylla might contained potential phenolic compounds that inhibiting the key enzyme in osteoarthritis development, which is the hyaluronidase enzyme through interruption of HYAL1 and HYAL1 gene expressions. The degradation of cartilage could also be inhibited by the plant through suppression of MMP-3 and MMP-13 protein expressions. We also reported that the inhibitory effect of Payena dasyphylla on hyaluronidase activity and expression might be due to its anti-oxidant property.

Diastereomers of lithospermic acid and lithospermic acid B from Monarda fistulosa and Lithospermum erythrorhizon

Monardic acids A (1) and B (2), which are (7R,8R) diastereomers of lithospermic acid (LA) and lithospermic acid B, respectively, were isolated from Monarda fistulosa. A (7S,8R) isomer (3) of LA was also isolated from this plant, and a (7R,8S) isomer (7) of LA was obtained from Lithospermum erythrorhizon. The absolute configuration of 1 was confirmed by analysis of its hydrolysates, 7-epiblechnic acid and 2R-3-(3,4-dihydroxyphenyl)-2-hydroxypropanoic acid. The configuration in the dihydrobenzofuran moieties of 2, 3, and 7 was extrapolated by using the phenylglycine methyl ester method and a Cotton effect at approximately 250–260nm in their electronic circular dichroism spectra. Diastereomers (1–3 and 7) displayed moderate hyaluronidase inhibitory and histamine release inhibitory activities.