Angelica Yinzi Alleviates Pruritus-Related Atopic Dermatitis through Skin Repair, Antioxidation, and Balancing Peripheral µ- and κ-opioid Receptors.

Background: Angelica Yinzi (AYZ) is a Chinese traditional herbal formula reported to attenuate itches and inflammation caused by atopic dermatitis (AD). However, the underlying mechanism of AYZ in the attenuation of itchiness and inflammation remains unknown. Objective: This study investigated the mechanism of AYZ in reducing itchiness in mice with 1-chloro-2,4-dinitrobenzene- (DNCB-)-induced atopic dermatitis. Methods: Hematoxylin and eosin (H&E) and toluidine blue staining were used to evaluate pathological changes in skin tissue, while an enzyme-linked immunosorbent assay (ELISA) was used to assess the cytokine levels in the skin. After that, qRT-PCR was performed to determine the mRNA levels of cytokines in the skin. Immunofluorescence and western blotting analysis were further used to assess µ-opioid receptor (MOR) expression and immunohistochemistry to assess the p-ERK, p-AKT, and κ-opioid receptor (KOR). Results: The AYZ treatment alleviated the AD clinical symptoms, including decreasing the scratching frequency, the ear thickness, and the infiltration of mast cells, lymphocytes, inflammatory cells, and mononuclear cells. In addition, AYZ inhibited the expression of interleukin (IL)-13, thymic stromal lymphopoietin (TSLP), and reduced neuraminidase (NA), corticotropin-releasing factor (CRF), and reactive oxygen species (ROS) expression. Markers involved in itches, such as p-ERK and p-AKT, were significantly downregulated following AYZ treatment. Besides, AYZ significantly increased MOR expression and downregulated KOR in the epidermis and spinal cord. Conclusion: Our findings imply that AYZ ameliorates pruritus-related AD through skin repair, antioxidation, and balancing peripheral MOR and KOR. The findings in this study lay a theoretical foundation for the control mechanism of peripheral itch.

Angelica Yinzi alleviates 1-chloro-2,4-dinitrobenzene-induced atopic dermatitis by inhibiting activation of NLRP3 inflammasome and down-regulating the MAPKs/NF-kB signaling pathway.

Background: Atopic dermatitis (AD), characterized by eczema as a chronic pruritic inflammatory skin disease, has become a serious health problem with recurrent clinical episodes. However, current clinical treatments have limited relief and are accompanied by adverse effects. Therefore, there is a necessity to develop new effective drugs for AD treatment. Angelica Yinzi (AYZ) is a classic ancient prescription for nourishing blood, moistening dryness, dispelling wind, and relieving itching. However, its mechanism for alleviating atopic dermatitis remains unknown. Therefore, this study aimed at determining the effects of AYZ and its potential mechanism in alleviating AD-like symptoms. Methods: In the present study, we used 1-chloro-2,4-dinitrobenzene (DNCB) to establish a mouse model of atopic dermatitis, where DNCB readily penetrates the epidermis to cause inflammation. Histopathological analysis was performed to examine the thickening of dorsal skin and infiltration in the inflammatory and mast cells in C57BL/6 mice. Additionally, the immunoglobulin E (IgE) levels in serum were determined by enzyme-linked immunosorbent assay (ELISA) kits. The IL-1ß and TNF-α expression were detected using qRT-PCR. Next, the Western blotting and immunohistochemistry assays were performed to assess the contribution of MAPKs/NF-κB signaling pathways and the NLRP3 inflammasome in AD responses. Results: Histopathological examination revealed that AYZ reduced the epidermal thickness of AD-like lesioned skin and repressed the infiltration of mast cells into AD-like lesioned skin. AYZ significantly decreased the phosphorylation of p38 MAPK, JNK, ERK and NF-κB and downregulated serum IgE levels and IL-1ß and TNF-α mRNA levels. Additionally, the NLRP3, ASC, Caspase-1, and IL-1ß expression in dorsal skin were effectively down-regulated following AYZ treatment (p < 0.05 and p < 0.01). Conclusion: These findings revealed that AYZ effectively suppressed AD-induced skin inflammation by inhibiting the activation of the NLRP3 inflammasome and the MAPKs/NF-kB signaling. Therefore, AYZ is a potential therapeutic agent against AD in the clinical setting.

Apoptosis induced by DICO, a novel non-aromatic B-ring flavonoid via a ROS-dependent mechanism in human colon cancer cells.

5,7-Dihydroxy-2-(1,2-isopropyldioxy-4-oxo-cyclohex-5-enyl)-chromen-4-one (DICO) is a novel non-aromatic B-ring flavonoid, isolated mainly from Macrothelypteris viridifrons and has anti-tumour properties. In this study, we investigated the cytotoxicity and underlying biochemical pathways leading to cell death, in response to DICO treatment of a human colon cancer cell line HT-29. Our results indicated that DICO induced apoptosis by elevating the generation of reactive oxygen species, which could be quenched by the antioxidants N-acetyl cysteine. In addition, activation of signal transducer and activator of transcription 3 and suppression of nuclear factor kappa B played a crucial role in DICO-induced apoptosis. Overall, our results provide mechanistic insights into the apoptotic action of a potential anti-tumour drug, DICO.

HPLC analysis, optimization of extraction and purification conditions, biological evaluation of total protoflavones from Macrothelypteris viridifrons.

The present study aims to evaluate phytochemical and pharmacological potential of total protoflavones from Macrothelypteris viridifrons. In the phytochemical study, an HPLC analysis method was established, and the optimal extraction and purification conditions were analyzed. The extractive condition was optimized as follows: the backflow extraction with 20 folds of 70% ethanol at 80◦C for 1 h twice. Moreover, by combining the alkali-extraction and acid-precipitation method with the macroporous resin purification technology, the final purity rate of total protoflavones was no less than 54.85%. In the pharmacological study, the total protoflavones from M. viridifrons showed a significant tumor-inhibitory effect in the H22 hepatoma cells transplantation model with a higher inhibitory rate of 55.76% in high dosage (100mg/kg) treatment group compared with the positive control group (20 mg/kg cyclophosphamide). Taken of all, these results support that protoflavones are the material basis of M. viridifrons as an anticancer folk medicine.

Determination of protoapigenone in rat plasma by high-performance liquid chromatography with UV detection and its application in pharmacokinetic studies.

A simple and sensitive HPLC method using UV detection was developed to determine the concentration of protoapigenone in rat plasma. Chromatographic separation was conducted on a C18 column with a mobile phase consisting of an acetonitrile-methanol-aqueous phase (containing 0.2% acetic acid, pH 3.0) system at a flow rate of 1.0 mL/min. The UV detector was set at 248 nm. The calibration curve was linear over the range of 0.031-10.0 µg/mL. The lower limit of quantification was 31 ng/mL. The recoveries for plasma samples ranged from 70.3 to 82.5%. The intra- and inter-day accuracy and precision fulfilled the international standards. This method was successfully applied to a pharmacokinetic study of protoapigenone in rats after oral administration of protoapigenone. It was shown that protoapigenone could be absorbed rapidly after oral administration and could reach the maximum concentration within 1 h.

DICO, a novel nonaromatic B-ring flavonoid, induces G2/M cell cycle arrest and apoptosis in human hepatoma cells.

DICO was a novel nonaromatic B-ring flavonoid obtained from Macrothelypteris torresiana. In the present work, we investigated the antitumor activity and the antineoplastic mechanism of DICO. Our study showed that DICO inhibited the growth of HepG2 cells in dose and time-dependent manners. As well as DICO induced G2/M cell cycle arrest and apoptosis via a ROS-mediated mitochondrial pathway. Western blot assay demonstrated that DICO decreased Bcl-2 level and induced Bax translocation to cause cytochrome c release. Subsequently, caspase-9 and caspase-3 were activated. Meanwhile, the alterations of cyclin A and B1, p-CDK1 and p-cdc25c levels were also observed in response to DICO treatment. Taken together, DICO displayed a significant antitumor effect through G2/M cell cycle arrest and apoptosis induction, which suggested DICO might have therapeutic potential against tumors.

Characterisation of phenols and antioxidant and hypolipidaemic activities of Lethariella cladonioides.

BACKGROUND: This study aimed to investigate the antioxidant and hypolipidaemic activities of an ethanol extract of Lethariella cladonioides (Nyl.) Krog (EE) and to characterise its chemical constituents. RESULTS: Nine phenols were identified as canarione, thamnolic acid, squamatic acid, vermicularin, norstictic acid, baeomycesis acid, lecanoric acid, barbatinic acid and usnic acid from analysis of EE by using high-performance liquid chromatography with a diode array detector-mass spectrometry. In antioxidant analysis in vitro, the highest scavenging rate of EEs on the 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals, superoxide anion, hydroxyl radicals and hydrogen peroxide was 81.55 ± 1.95%, 81.84 ± 4.00%, 74.28 ± 3.71% and 74.28 ± 3.71%, respectively. Meanwhile, after administration of EE for 6 weeks in high fat/cholesterol diet mice, the most significant reduction in levels of serum triglyceride, serum total cholesterol, serum low density lipoprotein cholesterol and liver malondialdehyde were 24%, 20%, 15% and 35%, respectively. The most significant increase in levels of serum high density lipoprotein cholesterol and liver superoxide dismutase was 35% and 88%, respectively. CONCLUSION: L. cladonioides possesses strong antioxidant and hypolipidaemic activities.

Anti-tumor and anti-angiogenic effects of Macrothelypteris viridifrons and its constituents by HPLC-DAD/MS analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Macrothelypteris viridifrons is widely distributed in south of China and has been used as folk medicine to treat cancer, hydropsy, and traumatic bleeding. AIM OF THE STUDY: To investigate the chemical constituents and the anti-tumor and anti-angiogenic effects of Macrothelypteris viridifrons. MATERIALS AND METHODS: An HPLC-DAD/MS technique was used to determine the flavonoid profile of Macrothelypteris viridifrons. The anti-tumor effect of Macrothelypteris viridifrons was evaluated by in vivo mice bearing H22 hepatoma cells transplantation tumor model. And the anti-angiogenic activity was investigated by measuring the effects on the in vitro proliferation, migration, and tube formation of human umbilical vein endothelial cells (HUVECs). Furthermore, the in vivo zebrafish model was applied to evaluate the anti-angiogenic effect of Macrothelypteris viridifrons. RESULTS: 18 flavonoids were identified from Macrothelypteris viridifrons. Administration of Macrothelypteris viridifrons significantly inhibited the tumor growth and the expression of vascular endothelial growth factor (VEGF) and CD34. Meanwhile, Macrothelypteris viridifrons showed significant inhibition on proliferation, migration and tube formation of HUVECs in vitro and the intersegmental vessels formation in zebrafish model. CONCLUSIONS: Macrothelypteris viridifrons showed significant anti-tumor and anti-angiogenic effects and might be developed as a novel anti-tumor drug.

A novel non-aromatic B-ring flavonoid: isolation, structure elucidation and its induction of apoptosis in human colon HT-29 tumor cell via the reactive oxygen species-mitochondrial dysfunction and MAPK activation.

The aim of the present study was to elucidate the chemical structure of a novel non-aromatic B-ring flavonoid (DHEC) isolated from Macrothelypteris viridifrons and to evaluate its putative molecular mechanism of action on induction of apoptosis in human colon HT-29 cancer cell. On the basis of MS, UV, IR, 1D and 2D NMR data, DHEC was identified as 2-(cis-1, 2-dihydroxy-4-oxo-cyclohex-5-enyl)-5-hydroxy-7-ethoxy-chromone. In addition, the cytotoxicity of DHEC and its effect on induction of apoptosis were confirmed by several assays. After treatment of HT-29 cell with DHEC, we observed the accumulation of intracellular reactive oxygen species, the loss of mitochondrial membrane potential, the alteration of expression of the Bcl-2 family members, the releasing of cytochrome c, the cleavage of poly (ADP-ribose) polymerase (PARP), and the activation of caspase-3, -8, and -9. Further analysis showed that the mitogen-activated protein kinase (MAPK) related proteins were stimulated by treatment with DHEC. These results suggest that DHEC exhibits potential anti-cancer activity in HT-29 cell through induction of apoptosis, which may highly be associated with reactive oxygen species-mitochondrial dysfunction as well as activation of MAPK signaling pathway.

[Flavonoids with special B-ring from Macrothelypteris viridifrons and their anti-proliferative effects on tumor cell].

OBJECTIVE: To study the chemical constituents of Macrothelypteris viridifrons and their anti-proliferative effects on tumor cell. METHOD: The compounds were isolated by column chromatography with silica gel, C18 reverse-phase silica gel, sephadex LH-20, and their structures were elucidated on the basis of physiochemical propertities and spectral analysis. The antitumor activities of all compounds were tested with MOLT4, Hep G2, A-549, MCF-7, HT-29, PC-3 tumor cell lines. RESULT: Five compounds were isolated and identified as protoapigenone (1), protoapigenin (2), protoapigenin-4'-O-beta-D-glucopyanoside (3), 5,7-dihydroxy-2-(1,2-isopropyldioxy-4-oxo-cyclohex-5-enyl) -chromen-4-one (4), 5,7-dihydroxy-2-(1-hydroxy-2,6-dimethoxy-cyclohex-4-oxo) -chromen-4-one (5), respectively. CONCLUSION: All compounds were obtained from this plant for the first time. Compounds 1, 4 and 5 showed strong anti-proliferative effects on six tumor cells, which were in concentration-dependent manner.

Flavonoids from the aerial parts of Macrothelypteris torresiana.

Two new flavone derivatives (1 and 2) were isolated from the aerial parts of Macrothelypteris torresiana, along with four known flavonoids: protoapigenin, apigenin, kaempferol and quercetin. The structures were determined on the basis of spectroscopic data. Compound 1 showed weak cytotoxic activity against human tumour cell lines HepG2 , MCF7 and K562.

Parathelypteriside attenuates cognition deficits in d-galactose treated mice by increasing antioxidant capacity and improving long-term potentiation.

Parathelypteriside (PG), a stilbenoid compound, was extracted from Parathelypteris glanduligera (kze.) ching that exhibits antioxidative and anti-inflammatory effects. The aim of this study was to investigate the protective effect of PG against the d-galactose (d-gal)-induced neurotoxicity in mice. It was found that long-term intraperitoneal (i.p.) injection of PG (5 or 10 mg/(kg day)) for two weeks significantly improved the behavioral performance of d-gal-treated mice in both Morris water maze test and step-down avoidance test. Biochemical examination revealed that PG reduced the increased levels of malondialdehyde (MDA), and attenuated the decreased activities of superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase in the hippocampus of d-gal-treated mice. Furthermore, the electrophysiological assay showed that PG significantly rescued the long-term potentiation (LTP) impairment in mice hippocampus, and western blotting analysis indicated that the effects of PG on LTP might be attributed to the activation of cAMP-response element-binding protein (CREB). Together, these results suggested that the natural product PG represented a potential source of medicine for the treatment of the neurodegenerative diseases.

Antioxidant, free radical scavenging, anti-inflammatory and hepatoprotective potential of the extract from Parathelypteris nipponica (Franch. et Sav.) Ching.

AIM OF THE STUDY: The present study was conducted to evaluate the antioxidant, free radical scavenging, hepatoprotective and anti-inflammatory potential of Parathelypteris nipponica (Franch. et Sav.) Ching. METHODS AND RESULTS: Antioxidant activity of the methanolic extract of Parathelypteris nipponica (Franch. et Sav.) Ching (TMPN) was studied using in vitro and in vivo models, total flavonoids content in TMPN was found to be 262 +/- 5.6 mg/g (w/w). The TMPN exhibited strong antioxidant activity with EC50 values in reductive ability (0.18 +/- 0.02 mg/ml) and ferric thiocyanate (FTC) assay (0.10 +/- 0.01 mg/ml), strong free radical scavenging activity as evidenced by the low EC50 values in DPPH (1,1-diphenyl-2-picrylhydrazyl) (2.00 +/- 0.02 mg/ml), superoxide anion (0.60 +/- 0.05 mg/ml), OH radicals (0.26 +/- 0.03 mg/ml), and hydrogen peroxide (0.45 +/- 0.03 mg/ml) methods. Acute toxicity study revealed that the LD50 value of the extract was more than the dose 2000 mg/kg bodyweight of mice. Hepatoprotective activity of TMPN was determined by the carbon tetrachloride (CCl4)-induced oxidative tissue injury in rat liver, the extract showed significant hepatoprotective activity that was evident by enzymatic examination and histopathological study. In assessing anti-inflammatory activity the carrageenan-induced rat paw oedema test was used, the extract reduced carrageenan-induced rat paw oedema in dose-dependent manner, achieving high degree of anti-inflammatory activity. CONCLUSION: This study provides a scientific basis for the ethnomedical claims that Parathelypteris nipponica (Franch. et Sav.) Ching is effective against inflammation and liver injury.

Antioxidant and hepatoprotective activity of ethanol extract of Arachniodes exilis (Hance) Ching.

AIM OF THE STUDY: The study was aimed to investigate the ethanol extract of Arachniodes exilis for the antioxidant and hepatoprotective activity. MATERIALS AND METHODS: Antioxidant activity was evaluated by different assays, including reducing power, lipid peroxidation, 2, 2'-diphenyl-1-picrylhydrazyl (DPPH), 2, 2'-azinobis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS), superoxide anion, hydroxyl radicals and hydrogen peroxide. The hepatoprotective activity of ethanol extract was studied on mice liver damage induced by CCL(4) by monitoring biochemical parameters. RESULTS: The extract showed potent activities on reducing power, lipid peroxide, DPPH, ABTS, superoxide anion, hydroxyl radical and hydrogen peroxide. And oral administration of Arachniodes exilis at different doses resulted in significant improvement on the levels of glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, malondialchehyche and superoxidedismutase. CONCLUSION: The results indicate that this plant possesses potential antioxidant and hepatoprotective properties and has therapeutic potential for the treatment of liver diseases.

[Phenols from the rhizomes of Arachniodes exilis].

OBJECTIVE: To study the chemical constituents of Arachniodes exilis. METHODS: The methanolic extract was isolated and purified by silica gel, C18 reverse phase silica gel and Sepadex LH-20 gel column chromatography. The chemical structures were elucidated on the basis of the physicochemical properties and spectral data. RESULTS: Eight phenolic compounds were obtained and identified as aspidin-BB (I), 4-methyl-2-butyl-3,5-dihydroxyphenol (II ), eriodictyol (III), epicatechin (IV), procyanidin B2 (V), miscathoside (VI), eriodictyol-7-O-beta-D-glucopyranuronide (VII), luteolin-4'-O-beta-D-glucopyranuronide (VIII), respectively. CONCLUSION: Compounds II-VIII are obtained from this plant for the first time.

Three new polyphenols from the rhizomes of Arachniodes exilis.

Three new polyphenols, araspidin BB (1), arachniodesin A (2) and arachniodesin B (3), together with two known compounds, epicatechin (4) and procyanidin B-2 (5), were isolated from the rhizomes of Arachniodes exilis. The structures of three new compounds were elucidated as 5-methyl-methylene-bis-phlorobutyrophenone (1), 4beta-ethoxycarbonylmethylepicatechin (2) and epicatechin-(4beta --> 8)-4beta-ethoxycarbonylmethylepicatechin (3), on the basis of their spectral analysis and by comparing them with the related model compounds. Compounds 4 and 5 were obtained from the title plant for the first time.

Chemical constituents of radix Ranunculus ternati.

3 Beta-acetoxy-(20S, 22E)-dammaran-22-en-25-ol, a new triterpene, was isolated along with five known triterpenes (ursolic acid, oleanolic acid, betulinic acid, 3-epiocotillol acetate, and dimmarenediol II acetate), and alpha-D-glc and sucrose from Radix Ranunculus ternati All of them, except oleanolic acid and alpha-D-glc, were isolated from the family of Ranunculaceae for the very first time, and the NMR data of sucrose was first described. In addition, the absolute configurations of alpha-D-glc and the glucose component of sucrose were determined.

Acetylated flavanone glycosides from the rhizomes of Cyclosorus acuminatus.

Six new flavanone glycosides (1-6) were isolated from the methanol extract of the rhizomes of Cyclosorus acuminatus, together with the parent flavanone glycoside 2a. Their structures were established on the basis of spectroscopic and chemical methods. All compounds showed moderate activity against Streptococcus pneumoniae and Haemophilus influenzae.

Flavan-4-ol Glycosides from the Rhizomes of Abacopteris penangiana.

Four new flavan-4-ol glycosides, abacopterins A-D (1-4), were isolated from a methanol extract of the rhizomes of Abacopteris penangiana, together with two known compounds, triphyllin A (5) and 6,8-dimethyl-7-hydroxy-4'-methoxyanthocyanidin-5-O-beta-D-glucopyranoside (6). Their structures were elucidated by extensive spectroscopic analysis and chemical methods. The cytotoxic activity of 1-5 against HepG2 human hepatoma cells was investigated.