Topical application of an ethanol extract prepared from Illicium verum suppresses atopic dermatitis in NC/Nga mice

Ethnopharmacological relevance

Illicium verum is a traditional herbal medicine with anti-inflammatory properties used in Asia. However, its usefulness in the treatment of allergic diseases remains unclear. This study evaluated the anti-inflammatory and antiallergic effects of I. verum extract (IVE) in a mouse model of atopic dermatitis.

Materials and methods

We investigated the effects of IVE on compound 48/80-induced histamine release, and phorbol 12-myristate13-acetate and calcium ionophore A23187-stimulated cytokines secretion in MC/9 mast cells. Atopic dermatitis was induced in NC/Nga mice by exposure to extract of house dust mite (Dermatophagoides farinae). After a topical application of IVE on ear and skin lesions, we evaluated the severity of skin symptoms, ear thickness, inflammatory cell infiltration, and serum levels of immunoglobulin E (IgE), histamine, interleukin (IL)-6, and intercellular adhesion molecule (ICAM)-1. In addition, we determined the expression of IL-4, IL-6, tumor necrosis factor (TNF)-α, interferon (IFN)-γ thymus- and activation-regulated chemokine (TARC), regulated on activation, normal T cell expressed and secreted (RANTES), ICAM-1, and vascular cell adhesion molecule (VCAM)-1 in ear tissues.

Results

IVE inhibited secretion of histamine, IL-4, IL-6, and TNF-α from mast cells in a dose-dependent manner. Topical application of IVE significantly reduced dermatitis scores, ear thickness, and serum levels of IgE, histamine, IL-6, and ICAM-1. Histopathological analysis demonstrated decreased epidermal thickening and dermal infiltration by inflammatory cells. In the ear lesions, IVE treatment reduced expression of IL-4, IL-6, TNF-α, TARC, RANTES, ICAM-1, and VCAM-1, but not IFN-γ.

Conclusions

These results indicate that IVE inhibits atopic dermatitis-like skin lesions by suppressing the expression of cytokines, chemokines, and adhesion molecules. These results suggest that IVE may be a potential therapeutic candidate for atopic dermatitis.     

Inhibitory effects of Cinnamomum cassia extract on atopic dermatitis-like skin lesions induced by mite antigen in NC/Nga mice

Aim of the study

: Cinnamomum cassia (C. cassia) has been traditionally used to treat allergic disease as well as dyspepsia, gastritis, and blood circulation disturbances. However, the antiallergic properties of C. cassia have not been fully verified using scientific tools. This study investigated the effectiveness of C. cassia extract (CCE) as an antiallergic agent in atopic dermatitis model and underlying mechanism.

Materials and methods

: The effect of CCE on mite antigen-treated NC/Nga mice was evaluated by examining skin symptom severity, levels of serum IgE, tumor necrosis factor-α (TNF-α), and histamine, skin histology, and mRNA expression of cytokines in the skin lesions. Moreover, the effect of CCE on TNF-α-and interferon-γ (IFN-γ)-induced chemokine production in human keratinocytes was investigated using ELISA.

Results

: CCE treatment of NC/Nga mice reduced the dermatitis score and the levels of serum IgE, histamine, and TNF-α. Histological examination showed inhibition of the thickening of the epidermis/dermis and reduced dermal infiltration of inflammatory cells. In skin lesions, mRNA expression of IL-4, TNF-α, and thymus and activation-regulated chemokine (TARC) was inhibited by CCE treatment. The production of TARC, macrophage-derived chemokine, and RANTES from IFN-γ-and TNF-α-stimulated human keratinocytes was suppressed by CCE treatment in a dose-dependent manner.

Conclusions

: CCE inhibits the development of atopic dermatitis-like skin lesions in NC/Nga mice by suppressing the T-helper 2 cell response.     

Inhibitory effects of Drynaria fortunei extract on house dust mite antigen-induced atopic dermatitis in NC/Nga mice

Ethnopharmacological relevance

Drynaria fortunei (Kunze) J. Sm has been widely used in traditional medicine for the treatment of inflammation, hyperlipidemia, arteriosclerosis, rheumatism, and bone healing. We investigated the anti-inflammatory effects of a 70% ethanol extract of Drynaria fortunei (DFE).

Materials and methods

We evaluated the anti-inflammatory effects of topically applied DFE on house dust mite Dermatophargoides farinae-induced atopic dermatitis-like skin lesions in NC/Nga mice.

Results

Treatment of NC/Nga mice with DFE reduced the dermatitis score, ear thickness, and serum levels of IgE, IgG1, and IL-6. Histopathological analyses of ear and skin lesions showed inhibition of the thickening of the epidermis and reduced epidermal/dermal infiltration of inflammatory cells. In ear lesions, mRNA expression levels of IL-4, IL-6, and tumor necrosis factor-α were reduced by DFE treatment.

Conclusions

DFE inhibited the development of dermatitis-like skin lesions in NC/Nga mice. These results suggest that DFE may be a therapeutic candidate for the treatment of AD.     

Effect of Alnus japonica extract on a model of atopic dermatitis in NC/Nga mice

Aim of the study

The bark of Alnus species has long been used in traditional oriental medicine in the treatment of many pathological conditions, including fever, hemorrhage, diarrhea, alcoholism, various skin diseases (e.g. chronic herpes, eczema and prurigo), and inflammation. In order to assess the immunomodulatory efficacy of a novel herbal medicine in treating atopic dermatitis, we measured serum levels of several allergic and inflammatory biomarkers in NC/Nga mice before and after treatment with this experimental agent.

Materials and methods

Gene and protein expression analyses of iNOS and COX-2 were quantified by real time PCR and Western blot analysis and serum levels of IL-4, -5 and -13 were also measured by ELISA, all of which were reduced after treatment with the experimental agent. Additionally, serum concentrations of IgE and blood eosinophil counts were reduced in treated mice.

Results

The topical application of leaf and bark extract from Alnus japonica suppressed the development of AD-like skin lesions. The percent of blood eosinophils was decreased after treatment with leaf and bark extract from Alnus japonica. The serum IgE and Th2-related cytokine levels were decreased after treatment with leaf and bark extract from Alnus japonica compared with those treated with base cream (vehicle treated AD group). The IL-4, IL-5 and IL-13 were lower than those of vehicle treated AD group.

Conclusions

We contend that leaf and bark extract from Alnus japonica may prove useful in the treatment of atopic dermatitis and other allergic skin diseases, although more in-depth clinical studies are necessary before clinical implementation.     

Inhibitory effects of Chelidonium majus extract on atopic dermatitis-like skin lesions in NC/Nga mice

Aim of the study

Chelidonium majus (CM) has traditionally been used for treatment of various inflammatory diseases including atopic dermatitis (AD). However its action on atopic dermatitis (AD) is unclear. Therefore, we investigated the effect of CM on AD using NC/Nga mice as an AD model.

Materials and methods

The effect of CM on 1-chloro-2,4-dinitrobenzene (DNCB) induced NC/Nga mice was evaluated by examining skin symptom severity, itching behavior, ear thickness, levels of serum immunoglobulin E (IgE), tumor necrosis factor-α (TNF-α), and interlukin-4 (IL-4), skin histology.

Results

The CM significantly reduced the total clinical severity score, itching behavior, ear thickness and the level of serum IgE in AD mouse model. CM not only decreased TNF-α but also IL-4.

Conclusion

These results suggest that CM may be a potential therapeutic modality for AD.     

Rehmannia glutinosa: review of botany, chemistry and pharmacology

Rehmannia glutinosa, a widely used traditional Chinese herb, belongs to the family of Scrophulariaceae, and is taken to nourish Yin and invigorate the kidney in traditional Chinese medicine (TCM) and has a very high medicinal value. In recent decades, a great number of chemical and pharmacological studies have been done on Rehmannia glutinosa. More than 70 compounds including iridoids, saccharides, amino acid, inorganic ions, as well as other trace elements have been found in the herb. Studies show that Rehmannia glutinosa and its active principles possess wide pharmacological actions on the blood system, immune system, endocrine system, cardiovascular system and the nervous system. Currently, the effective monomeric compounds or active parts have been screened for the pharmacological activity of Rehmannia glutinosa and the highest quality scientific data is delivered to support the further application and exploitation for new drug development.     

Effects of Catalpa ovata stem bark on atopic dermatitis-like skin lesions in NC/Nga mice

Ethnopharmacological relevance

The stem bark of Catalpa ovata has been used as a traditional herbal medicine for the treatment of various inflammatory diseases such as itching and scabies.

Aim of the study

In the present study, we investigated the anti-AD effects of Catalpa ovata stem bark on Dermatophagoides farinae-induced AD in a NC/Nga mouse AD model. We determined dermatitis score, histology, IgE, cytokines, and chemokines related to hypersensitive immune responses in AD. The mechanism of action was also investigated using HaCaT cells.

Materials and methods

We investigated the topical effects of Catalpa ovata stem bark on AD-like skin lesions in NC/Nga mice. Five category-experiments were performed, including assessment of dermatitis score; histological analysis of dorsal skin lesions; quantitative measurement of serum total IgE; quantitative measurement of cytokines (IL-1β, IL-4, IL-5, IL-6, IL-13, TNF-α) from dorsal tissue; and RT-PCR analysis of for TSLP and TARC mRNA expression in HaCaT cells.

Results

The clinical dermatitis score was significantly lower in Catalpa ovata extract (COE) groups than in the control group. Histological analysis showed that COE inhibited hypertrophy and hyperkeratosis of the epidermis, intracellular edema, and reduced the infiltration of inflammatory cells. COE significantly inhibited serum total IgE; Th2 cytokines IL-4, IL-5 and IL-13; pro-inflammatory cytokines IL-1β, IL6 and TNF-α; the Th2 chemokine TARC and the pro-Th2 cytokine TSLP.

Conclusion

These results demonstrate that Catalpa ovata stem bark may be a useful external medicine for treatment of AD. Further investigation is necessary to determine appropriate COE dosage and to evaluate the safety of this medicinal herb.     

地黄块根膨大发生和驱动的组织观察及激素相关基因的调控分析

目的:为了研究地黄块根膨大的发育特征,揭示激素相关基因在这个过程中的调控作用机制。方法:该研究以地黄品种"温-85"为实验材料,采集不同发育时期的地黄块根进行表型观察,利用半薄树脂切片技术观察块根的组织形态。从地黄转录组数据库中挑选与激素合成和响应相关的基因,利用qRT-PCR技术研究激素相关基因在地黄不同发育时期块根组织中的表达变化。结果:地黄的发育阶段可划分为苗期、拉线期、块根膨大初期,膨大中期,膨大后期和成熟期。块根解剖学特征分析表明次生形成层的分裂启动了地黄块根膨大的进程;膨大前期次生形成层的持续快速分裂和副形成层的形成驱动了块根的向外(平周)和侧向(圆周)持续快速膨大。在半薄和超薄切片观察过程中发现在地黄块根中存在有大量的油脂细胞。定量结果分析显示IAA,CK,ABA,乙烯,JA,EB的合成和响应基因整体呈现上调表达趋势,GA的合成与响应基因呈下调趋势,其负调控因子基因则上调表达。大部分激素合成相关基因的表达高峰期处在拉线期、膨大前期。结论:拉线期和块根膨大前期是地黄块根发育最为关键的时期,IAA,CK,ABA,乙烯,JA,EB的合成和响应基因的上调与GA合成基因的下调能促进块根的发育。油脂细胞可能与地黄药用成分的合成与储存有密切关系。该研究为阐述地黄块根发育的规律和分子调控机制进行了有益的探索。     

Effects of Scutellariae radix and Aloe vera gel extracts on immunoglobulin E and cytokine levels in atopic dermatitis NC/Nga mice

Aim of the study

The present study aimed to investigate the effects of Scutellariae radix (SR) and Aloe vera gel (AV), alone or in combination, on levels of immunoglobulin E (IgE) and inflammatory cytokines in spontaneous atopic dermatitis(AD)-like skin lesions.

Materials and methods

After spontaneous AD-like skin lesion was developed by adaptation to conventional conditions, mice were randomly assigned to control, SR (50 mg/kg, p.o.), AV (0.8 mg/kg, p.o.) and SRAV (50 mg of SR and 0.8 mg of AV/kg, p.o.) groups, and were treated for 6 weeks.

Results

SR and SRAV suppressed IL-5 levels compared with control, but had no effects on IgE levels (P < 0.05). AV increased IgE levels, but decreased both IL-5 and IL-10 compared with control (P < 0.05).

Conclusion

These results suggest that SR and AV modulate immunological responses in AD, mainly through influencing IL-5 or IL-10 levels.     

The bark of Betula platyphylla var. japonica inhibits the development of atopic dermatitis-like skin lesions in NC/Nga mice

The bark of Betula platyphylla Sukat. var. japonica Hara (Asian White Birch, AWB) is one of the herbs used in Eastern countries for the treatment of various inflammatory diseases including atopic dermatitis (AD). The present study was performed to examine if and how the bark of AWB inhibits the development of AD-like skin lesions in NC/Nga mice induced by repeated application of picryl chloride (PC). With this aim, we examined the skin symptom severity, itching behavior, serum immunoglobulin (Ig) E level and mRNA expression of cytokines at iliac and cervical lymph nodes in the mice. Oral administration of AWB extracts (25, 100 and 250 mg/kg) to the PC-treated mice inhibited the development of AD-like skin lesions as exemplified by a significant decrease in the total skin severity scores, itching behavior and a decrease in hypertrophy and infiltration of inflammatory cells into dermis. The serum IgE level was also significantly reduced by AWB extract. In the RT-PCR results, the expression of interleukin-4 mRNA was reduced by AWB extract, whereas the expression of interferon-gamma mRNA was not changed. These results suggest that AWB inhibits the development of AD-like skin lesions in NC/Nga mice through the suppression of the T-helper 2 cell response.     

Korean red ginseng extract ameliorates skin lesions in NC/Nga mice: an atopic dermatitis model

Aim of the study

Korean red ginseng (KRG, Panax ginseng C.A. Meyer) has traditionally been considered to harbor anti-allergic effects, however its action on atopic dermatitis (AD) is unclear. Therefore, we investigated the effect of KRG on AD using NC/Nga mice as an AD model. In addition, we examined the effect of aprepitant (substance P specific neurokinin receptor antagonist) on AD.

Materials and methods

The KRG extract and aprepitant were administered orally to NC/Nga mice. The efficacy of KRG and aprepitant was evaluated by assessing total clinical severity score, ear thickness, serum IgE level and histology. In addition, mRNA and protein expression were measured by real-time RT-PCR and immunohistochemistry, respectively.

Results

The KRG extract significantly reduced the total clinical severity score, ear thickness and the level of serum IgE in AD mouse model, whereas aprepitant reduced only the serum IgE level. KRG not only decreased TNF-α, IFN-γ and substance P but also reduced the infiltration of FOXP3+ regulatory T (Treg) cells and CD1a+ Langerhans cells in the lesions, whereas aprepitant decreased only substance P and the infiltration of Treg cells.

Conclusion

These results suggest that KRG extract may be a potential therapeutic modality for AD and aprepitant could be used as adjunctive agent to control pruritus in AD.     

地黄资源遗传多样性分析

目的:研究不同地区栽培地黄及野生地黄品种间ITS序列差异,为地黄的系统发育和分子鉴定提供依据。方法:用CTAB法提取地黄叶片中总DNA,对PCR产物进行克隆测序,采用MEGA4.0进行系统进化分析。结果:各供试地黄的ITS区序列总长度为613~614 bp,长度变异仅1 bp,其中ITS1长度为224~225 bp,G+C占60.4%~63%;ITS2长度为224~225 bp,G+C占57.1%~65.3%;5.8S长度高度保守,均为164 bp。系统发育分析表明:北京2号地黄与其他地黄资源差异性较大,亲缘关系较远。野生地黄居群内差异较小,居群间没有差异。河南栽培地黄与山西、山东省栽培地黄间没有差异;野生地黄资源中神农山和青天河一带地黄与山东省栽培地黄亲缘关系最近,而河南栽培地黄和山西省栽培地黄与野生地黄没有差异。结论:不同地区地黄资源种间亲缘关系较近,系统分化不明显。     

怀地黄不同主栽品种叶形态特征比较

目的:通过观察怀地黄不同主栽品种叶的形状,非腺毛和气孔器大小、密度等特征,为怀地黄主栽品种的鉴定及优良品种的选育提供依据。方法:用直尺测量叶片的长度和宽度,在扫描电镜下计数不同主栽品种怀地黄叶表面非腺毛和气孔的密度,测量其长度。结果:叶片长度、宽度及长宽比,非腺毛和气孔的长度、密度等,在怀地黄不同主栽品种间具有差异。结论:叶的形状,非腺毛和气孔的密度、长度等叶的形态特征可以作为怀地黄不同品种间鉴别的依据。     

The fruits of Cudrania tricuspidata suppress development of atopic dermatitis in NC/Nga mice

The fruits of Cudrania tricuspidata are a medicinal herb in Korea, known for its antiatherosclerotic and antiinflammatory effects. Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by the influx of lymphocytes into the dermis. Using an animal model of AD, we assessed whether C. tricuspidata suppresses the development of AD-like skin lesions. Cudrania tricuspidata was administered orally to NC/Nga mice with Dermatophagoides-farinae-induced AD-like lesions for 49 days. The effects of C. tricuspidata were assessed by measuring clinical symptoms, swelling of the skin on the back and ears, and plasma concentrations of mTARC (mouse thymus and activation regulated chemokine), histamine and immunoglobulin E (IgE). We found that C. tricuspidata (60 mg/kg/day) inhibited the development of AD-like skin lesions, reduced skin dermatitis scores and inhibited the histological changes induced by repeated application of D. farinae. In addition, C. tricuspidata inhibited the increases in plasma concentrations of mTARC, histamine and IgE induced by D. farinae. These findings indicate that C. tricuspidata inhibits the development of AD-like skin lesions induced by repeated applications of D. farinae in sensitized NC/Nga by suppressing plasma concentrations of mTARC, histamine and IgE.     

地黄生长期根叶中梓醇及总环烯醚萜苷含量变化

地黄主要活性成分为环烯醚萜苷,其中含量最高的是梓醇。该文主要对地黄生长发育过程中根叶所含总环烯醚萜苷和梓醇的含量变化进行研究。以地黄85-5为材料,于地黄不同生长期进行不同直径块根、不同叶龄的样品采集,分别采用紫外分光光度法和高效液相色谱法测定总环烯醚萜苷和梓醇的含量。研究结果表明,7—9月同一生长期地黄不同叶龄中总环烯醚萜苷和梓醇含量,幼嫩组织中含量较高,随着叶龄的增加而逐步下降;比较各生长期相同着生部位的叶片中梓醇和总环烯醚萜苷含量,大多数部位的叶片在10月达到最高;同一生长期块根直径越大,有效成分含量越高;梓醇含量同总环烯醚萜苷含量呈正相关。以期通过以上初步结论可以为地黄的萜类代谢调控研究和栽培技术提供参考依据。     

地黄叶片试管块根诱导体系优化的研究

目的:研究不同浓度蔗糖和植物生长物质对地黄叶片试管块根诱导的影响,建立从地黄叶片诱导试管块根的高效体系.方法:以85-5地黄试管苗的叶片作为外植体,先经生根诱导,后转接至正交设计的培养基中,诱导地黄试管块根.结果与结论:NAA对试管地黄诱导影响显著,其次分别为蔗糖和6-BA,以试管苗叶片为外植体诱导地黄试管块根的最佳培养基为MS+6-BA3 mg·L-1+NAA0.1 mg·L-1+蔗糖70g·L-1.该研究为今后利用地黄试管块根进行人工种子和次生代谢研究提供了高效的培养体系.     

应用HPLC指纹图谱技术确定熟地黄炮制终点

目的:研究熟地黄炮制过程中HPLC指纹图谱的变化及确定炮制终点。方法:用HPLC指纹图谱技术检测生地黄炮制成熟地黄过程中主要活性成分梓醇、5-羟甲基糠醛(5-HMF)、麦角甾苷等及指纹的变化,并通过与标准熟地黄指纹图谱的比较寻找最佳加工时间。结果:在炮制过程中梓醇快速下降至几乎消失,5-HMF呈直线上升趋势,而麦角甾苷变化不明显,生地黄在蒸制26h后所制样品与标准熟地黄相似性最大,因此26h是熟地黄清蒸蒸制的最佳时间。结论:指纹图谱有利于显示样品成分变化及精确控制加工过程,可有效用于熟地黄加工过程中的质量控制。