Alpinetin Research Articles

Alpinetin ameliorates bleomycin-induced pulmonary fibrosis by repressing fibroblast differentiation and proliferation

ObjectiveIdiopathic pulmonary fibrosis (IPF) is a progressive and irreversible interstitial lung disease with a poor prognosis. Alpinetin (ALP), derived from Alpinia katsumadai Hayata, has shown potential as a therapeutic measure of various diseases. However, the utilization of ALP in managing pulmonary fibrosis and its underlying mechanisms are still not fully understood. MethodsA well-established mouse model of pulmonary fibrosis induced by bleomycin (BLM) was used in this study. The antifibrotic effects of ALP on histopathologic manifestations and expression levels of fibrotic markers were examined. Subsequently, the impact of ALP on fibroblast differentiation, proliferation, apoptosis, and associated signaling pathways was investigated to elucidate the underlying mechanisms. ResultsIn the present study, we observed that ALP effectively mitigated BLM-induced pulmonary fibrosis in mice, as evidenced by histopathological manifestations and the expression levels of fibrotic markers. Furthermore, the in vitro experiments demonstrated that ALP treatment attenuated the ability of fibroblasts to differentiate into myofibroblasts. Mechanically, our findings provided evidence that ALP suppressed fibroblast-to-myofibroblast differentiation by repressing TGF-β/ALK5/Smad signaling pathway. ALP was found to possess the capability of inhibiting fibroblast proliferation and promoting apoptosis of fibroblasts induced by TGF-β. ConclusionIn general, ALP may exert therapeutic effects on pulmonary fibrosis by modulating the differentiation, proliferation, and apoptosis of fibroblasts. Although its safety has been demonstrated in mice, further studies are required to investigate the efficacy of ALP in treatment of patients with IPF.

Synergistic Inhibitory Effect of Multiple Polyphenols from Spice on Acrolein during High-Temperature Processing.

Acrolein (ACR) is a toxic unsaturated aldehyde that is produced during food thermal processing. Here, we investigated the synergistic effect of polyphenols in binary, ternary, and quaternary combinations on ACR by the Chou-Talalay method, and then explored the synergistic effect of cardamonin (CAR), alpinetin (ALP), and pinocembrin (PIN) in fixed proportion from Alpinia katsumadai Hayata (AKH) combined with curcumin (CUR) in the model, and roasted pork using LC-MS/MS. Our results showed that their synergistic effect depended on the intensification of their individual trapping ACR activities, which resulted in the formation of more ACR adducts. In addition, by adding 1% AKH (as the carrier of CAR, ALP, and PIN) and 0.01% CUR (vs. 6% AKH single) as spices, more than 71.5% (vs. 54.0%) of ACR was eliminated in roast pork. Our results suggested that selective complex polyphenols can synergistically remove the toxic ACR that is produced in food processing.

Exploration of Pharmacological Potential of Alpinetin

Abstract: Secondary metabolites found in plants are a natural source of bioactive chemicals. These secondary metabolites are vital for the survival of plants and have a number of medicinal properties, which can be utilised to treat human illnesses. Alpinetin (ALP) is one of the secondary metabolites which belongs to the Flavonoid category of phytochemicals and is present in Amomum subulatum Roxb's. Alpinetin has been found to possess antioxidant, anti-inflammatory, anticancer, hepatoprotective and renoprotective activity, along with several other biological properties. This review is focused on the exploration of the pharmacological activities of Alpinetin. ALP is considered a prospective candidate for future clinical investigations due to the number of therapeutic properties.

Alpinetin Suppresses Zika Virus-Induced Interleukin-1β Production and Secretion in Human Macrophages.

Zika virus (ZIKV) infection has been recognized to cause adverse sequelae in the developing fetus. Specially, this virus activates the excessive release of IL-1β causing inflammation and altered physiological functions in multiple organs. Although many attempts have been invested to develop vaccine, antiviral, and antibody therapies, development of agents focusing on limiting ZIKV-induced IL-1β release have not gained much attention. We aimed to study the effects of alpinetin (AP) on IL-1β production in human macrophage upon exposure to ZIKV. Our study demonstrated that ZIKV stimulated IL-1β release in the culture supernatant of ZIKV-infected cells, and AP could effectively reduce the level of this cytokine. AP exhibited no virucidal activities against ZIKV nor caused alteration in viral production. Instead, AP greatly inhibited intracellular IL-1β synthesis. Surprisingly, this compound did not inhibit ZIKV-induced activation of NF-κB and its nuclear translocation. However, AP could significantly inhibit ZIKV-induced p38 MAPK activation without affecting the phosphorylation status of ERK1/2 and JNK. These observations suggest the possibility that AP may reduce IL-1β production, in part, through suppressing p38 MAPK signaling. Our current study sheds light on the possibility of using AP as an alternative agent for treating complications caused by ZIKV infection-induced IL-1β secretion.

Alpinetin alleviates osteoporosis by promoting osteogenic differentiation in BMSCs by triggering autophagy via PKA/mTOR/ULK1 signaling.

Osteoporosis, a systemic bone disease that is characterized by a reduction in bone mass and destruction of bone microstructure, is becoming a serious problem worldwide. Bone marrow mesenchymal stem cells (BMSCs) can differentiate into bone-forming osteoblasts, and play an important role in maintaining homeostasis of bone metabolism, thus being a potential therapeutic target for osteoporosis. Although the phytochemical alpinetin (APT) has been reported to possess a variety of pharmacological activities, it is still unclear whether APT can influence the osteogenic differentiation of on BMSCs and if it can improve osteoporosis. In this study, we found that APT treatment was able to enhance osteogenic differentiation levels of human BMSCs in vitro and mouse ones in vivo as revealed by multiple osteogenic markers including increased alkaline phosphatase activity and osteocalcin expression. Mechanistically, the protein kinase A (PKA)/mTOR/ULK1 signaling was involved in the action of APT to enhance the osteogenic differentiation of BMSCs. In addition, oral administration of APT significantly mitigated the bone loss in a dexamethasone-induced mouse model of osteoporosis through strengthening PKA signaling and autophagy. Altogether, these data demonstrate that APT promotes osteogenic differentiation in BMSCs by augmenting the PKA/mTOR/ULK1 autophagy signaling, highlighting its potential therapeutic application for treating osteoporotic diseases.

Alpinetin Suppresses Effects of TGF-β1 on Stimulating the Production and Organization of Fibrotic Markers in Human Primary Dermal Fibroblasts.

Overgrowths of dermal fibroblasts and myofibroblast phenoconversion in response to TGF-β stimulation are the hallmarks of skin fibrosis. Constitutive activation of dermal fibroblasts by TGF-β induces the excessive production of extracellular matrix as well as certain key intracellular proteins which form a complex interaction network. Current therapies include monoclonal anti-bodies against TGF-β and surgery, but these treatments generally elicit a limited effect on certain kinds of skin fibrosis. In the current study, we investigated the effects of alpinetin (AP) on human primary dermal fibroblasts (HPDFs) stimulated with TGF-β1. Results demonstrated that AP exhibited strong inhibitory effects on TGF-β1-induced proliferation and migration of HPDFs. AP also inhibited TGF-β1-induced morphological changes of fibroblasts to myofibroblasts, and these were found to be from its effects on blocking actin stress fiber formation and organization. The expression of major fibrotic molecules including α-SMA and type I collagen upon TGF-β1 stimulation was also inhibited by AP. In addition, AP attenuated TGF-β1-induced production and organization of vimentin, β-catenin, and N-cadherin, important for the pathophysiology of skin fibrosis. In conclusion, we revealed that AP has an ability to reverse the fibrotic effects of TGF-β1 at the cellular level, and this discovery suggests the therapeutic potential of AP for skin fibrosis.

Alpinetin promotes hair regeneration via activating hair follicle stem cells

BackgroundAlopecia affects millions of individuals globally, with hair loss becoming more common among young people. Various traditional Chinese medicines (TCM) have been used clinically for treating alopecia, however, the effective compounds and underlying mechanism are less known. We sought to investigate the effect of Alpinetin (AP), a compound extracted from Fabaceae and Zingiberaceae herbs, in hair regeneration.MethodsAnimal model for hair regeneration was mimicked by depilation in C57BL/6J mice. The mice were then topically treated with 3 mg/ml AP, minoxidil as positive control (PC), or solvent ethanol as vehicle control (VC) on the dorsal skin. Skin color changes which reflected the hair growth stages were monitored and pictured, along with H&E staining and hair shaft length measurement. RNA-seq analysis combined with immunofluorescence staining and qPCR analysis were used for mechanism study. Meanwhile, Gli1CreERT2; R26RtdTomato and Lgr5EGFP−CreERT2; R26RtdTomato transgenic mice were used to monitor the activation and proliferation of Gli1+ and Lgr5+ HFSCs after treatment. Furthermore, the toxicity of AP was tested in keratinocytes and fibroblasts from both human and mouse skin to assess the safety.ResultsWhen compared to minoxidil-treated and vehicle-treated control mice, topical application of AP promoted anagen initiation and delayed catagen entry, resulting in a longer anagen phase and hair shaft length. Mechanistically, RNA-seq analysis combined with immunofluorescence staining of Lef1 suggested that Lgr5+ HFSCs in lower bulge were activated by AP via Wnt signaling. Other HFSCs, including K15+, Lef1+, and Gli1+ cells, were also promoted into proliferating upon AP treatment. In addition, AP inhibited cleaved caspase 3-dependent apoptosis at the late anagen stage to postpone regression of hair follicles. More importantly, AP showed no cytotoxicity in keratinocytes and fibroblasts from both human and mouse skin.ConclusionThis study clarified the effect of AP in promoting hair regeneration by activating HFSCs via Wnt signaling. Our findings may contribute to the development of a new generation of pilatory that is more efficient and less cytotoxic for treating alopecia.

Alpinetin: a Dietary Flavonoid with Diverse Anticancer Effects.

Cancer is a global burden and mechanistically complex disease with a plethora of genetic, physiological, metabolic, and environmental alterations. The development of dietary nutraceuticals into cancer chemotherapeutics has emerged as a new paradigm in cancer treatment. Alpinetin (ALPI) is a novel flavonoid component of multiple edible and medicinal plants and possesses a wide range of biological and pharmacological activities including antibacterial, anti-hemostatic, anti-oxidative, anti-hepatotoxic, stomachic, immunosuppressive, and anti-inflammatory. Recently, ALPI has been reported as a bioactive dietary nutraceutical with promising anticancer activity in various human cancers through multiple mechanisms. The purpose of this review is to compile the data on natural sources of ALPI, and its anticancer activity including cellular targets and anticancer mechanism in various human cancers. Moreover, this review will set the stage for further design and conduct pre-clinical and clinical trials to develop ALPI into a lead structure for oncological therapy.

Interconversion and Acrolein-Trapping Capacity of Cardamonin/Alpinetin and Their Metabolites In Vitro and In Vivo.

People are at high risk of exposure to endogenous and exogenous acrolein (ACR). ACR can cause a multitude of illnesses, including cardiovascular disease, Alzheimer's disease, and diabetes. In this study, we investigated the reaction pathway of cardamonin (CAR) or alpinetin (ALP) with ACR and the interconversion of CAR and ALP in vitro at 37 °C in phosphate-buffered saline using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Subsequently, ACR adducts of CAR, ALP, and their metabolites, for example, CAR-ACR-1, ALP-ACR, mono-ACR-pinocembrin chalcone (PIN-ACR), and mono- and di-ACR-naringenin (NAR-ACR and NAR-2ACR), were detected in urine samples, but only CAR-ACR-1 and ALP-ACR were detected in fecal samples from the CAR- and ALP-treated mouse groups using ultraperformance liquid chromatography-MS/MS, respectively. Quantitative analyses showed that CAR, ALP, and their metabolites markedly scavenged ACR in a dose-dependent manner in vivo. Furthermore, we also found that the metabolites of CAR or ALP remained and promoted the ACR-trapping ability.

Alpinetin delays high-fat diet-aggravated lung carcinogenesis.

Alpinetin (ALP) has been reported to act as an anticancer agent. This study was carried out to elucidate the effect of ALP on high-fat diet (HFD)-induced aggressive cancer progression. C57BL/6 mice were fed with a control diet (CD) or HFD and administered with ALP. Following 6weeks of feeding, mice were inoculated subcutaneously with Lewis lung carcinoma cells (LLC) to develop transplanted lung tumour. ALP suppressed cell proliferation which drives HFD-induced lung cancer progression. ALP inhibited lipid accumulation in tumour and tumour cells cultured in vitro. qPCR and ELISA analysis of tumour tissues revealed ALP restrained macrophages accumulation, M2s polarization and chemokine secretion. Further, in macrophages cultured in tumour cells conditioned medium (CM), ALP was confirmed to decrease M2s markers expression and chemokine production under high fat. These results demonstrate that ALP suppresses HFD-promoted harmful changes in tumour microenvironments which are crucial in curbing pulmonary tumour aggravation.

Alpinetin Protects Chondrocytes and Exhibits Anti-Inflammatory Effects via the NF-κB/ERK Pathway for Alleviating Osteoarthritis.

Osteoarthritis (OA) is a chronic degenerative joint disease which is greatly affected by the inflammatory response triggered by the NF-κB signaling pathway. Alpinetin (APT) is a natural flavonoid compound, which has been reported to have many important biological activities such as antibacterial, antitumor, and anti-inflammatory. However, the action of its effect on chondrocytes in OA has yet to be elucidated. In this study, we investigated APT's anti-inflammatory action. The effects of APT on cell viability and cytotoxicity of rat chondrocytes was investigated by CCK8. Western blotting, qRT-PCR, and immunofluorescent staining were used to elucidate the molecular mechanisms and signaling pathways of APT mediating anti-inflammatory effects on chondrocytes. An OA model was induced by destabilization of the medial meniscus (DMM) in rats, then APT was injected into the knee articular cavity to examine its anti-inflammatory effects in vivo. These results showed that APT could reduce the TNF-α-induced increase of MMP-13 and ADAMTS-5 and decrease of COL2A1 levels. APT antagonized TNF-α-induced down-regulation of BCL-2 and CDK1. Further studies have shown that APT simultaneously repressed cell nucleus translocation of p65 and the phosphorylation of IκB and activated the phosphorylation of ERK. In vivo, APT suppressed cartilage matrix degradation. In conclusion, APT appears to favorably modulate anti-inflammatory effects in chondrocytes making it a promising compound for OA treatment. Graphical Abstract Inhibitory effects of Alpinetin on TNF-α-induced NF-κB activation resulted in destruction of cartilage in rat OA chondrocytes in vitro. The TNF-α receptor were stimulated by TNF-α, activating the cytoplasmic IκBα kinases(IKKS), then IKKs will be phosphorylated, and subsequently degraded by the ubiquitin-proteasome system. NF-κB transfer to the nucleus and bind various NF-κB regulates the NF-κB recognition site in the promoter region. Which triggers the gene expression of pro-inflammatory and pro-apoptotic. However, Alpinetin couldinhibits the NF-κB signaling pathway in different ways: APT inhibits IκBα phosphorylation, preventing phosphorylated ubiquitination of IκBα further. Moreover, APT blocks translocation of the activated NF-κB to the nucleus, to protect the cartilage tissue from damage.

Alpinetin Inhibits Oral Squamous Cell Carcinoma Proliferation via miR-211-5p Upregulation and Notch Pathway Deactivation

Background: The effect of alpinetin (ALP) on miR-211-5p level and function in oral squamous cell carcinoma (OSCC) remains unclear.Materials and methods: Human OSCC cell lines (CAL-27 and TCA-8113) and a mouse xenograft model with subcutaneously injected TCA-8113 cells were used. Effect of ALP treatment on cell viability, cell cycle distributions, and p-p53, p21, c-PARP, cyclin D1, NICD, HES1, and miR-211-5p expression levels was analyzed. Influence of ALP on tumor volume and weight was determined.Results: ALP treatment (at doses 400 and 500 µM) significantly decreased the viability of CAL-27 and TCA-8113 cells (P < 0.05). It upregulated the number of cells in G1 phase and miR-211-5p expression, increased p-p53, p21, and c-PARP levels, and decreased cyclin D1 levels. Furthermore, miR-211-5p mimic treatment increased the number of cells in G1 phase, and p53, p21, and c-PARP levels, and decreased cyclin D1 levels. Contrasting effects were observed under anti-miR-211-5p treatment. ALP downregulated NICD and HES1, whereas anti-miR-211-5p increased NICD and HES1 expression. ALP effects were alleviated in both cell lines under Jagged-1 overexpression plasmid treatment. Finally, ALP inhibited tumor growth and increased miR-211-5p expression in vivo.Conclusion: ALP-induced miR-211-5p upregulation and Notch pathway deactivation may be involved in its anti-proliferative effects in OSCC.

Determination of the raw material feeding (Gentiana scabra, Katsumade Galangal Seed and dried tangerine peel) in compound gentian tincture by HPLC-MS/MS

Objective To establish a HPLC-MS/MS method for comprehensive monitor and control of the raw material feeding (Gentiana scabra, Katsumade Galangal Seed and dried tangerine peel), and determination of Gentiopicroside, Alpinetin, Cardamonin and Hesperidin in Compound gentian tincture. Methods The separation was performed on an Inertsil ODS-3 (4.6 mm×150 mm, 5 μm) analytical column with the mobile phase consisting of acetonitrile-0.1% formic acid solution by gradient elution program, and the column temperature was 40 ℃. Active ingredients were separated by HPLC. The Electrospray Ionization Mass (ESI) source was applied and operated in the negative ion mode, and reactions ion monitoring mode (MRM) for quantitative analysis were selected. Results Through the analysis of the samples with mixed extract the same characteristic peak in MS was found to determine the proprietary Chinese medicine according to the prescription feeding process. The calibration curve of Gentiopicroside, Alpinetin,Cardamonin and Hesperidin were linear: 103.26-619.56 μg (r=0.999 0), 109.50-657.00 μg (r=0.999 5), 105.50-633.00 μg (r=0.996 9), 105.02-630.12 μg (r=0.999 5). The precision was Gentiopicroside 0.81%, Alpinetin 0.48%, Cardamonin 0.61% and Hesperidin 1.55% respectively. The average recovery rate were Gentiopicroside 95.08%, Alpinetin 93.28%, Cardamonin 94.78% and Hesperidin 95.04% respectively. Conclusions The method was proved to be simple, accurate, reliable, high sensitivity and can be used for determination and control of the raw material feeding (Gentiana scabra, Katsumade Galangal Seed and dried tangerine peel). Key words: Gentiopicroside; Alpinetin; Cardamonin; Hesperidin; Compound gentian tincture; HPLC-MS/MS

Alpinetin improved high fat diet-induced non-alcoholic fatty liver disease (NAFLD) through improving oxidative stress, inflammatory response and lipid metabolism

The non-alcoholic fatty liver disease (NAFLD) has become a serious medical problem and an increasing threat to public health. It is characterized by the abnormal fat accumulation in liver without excessive alcohol intake. The concurrent NAFLD might up-regulate the risk of chronic kidney disease as well as the mortality rate. Though various drugs have been investigated to attenuate NAFLD, further study is still necessary to find new therapeutic strategy and to reveal the underlying molecular mechanism. In the present study, NAFLD animal models were induced by feeding with high fat (HF) diet for 8 weeks. Alpinetin (ALP) was given to mice for another 8 weeks together with HF. Hepatic and renal function, oxidative stress, inflammatory response and lipid metabolism were calculated. And human liver cells of HL-7702 were cultured with high fructose (5mM) with or without ALP. The findings indicated that ALP down-regulated lipid accumulation in liver tissue samples. The higher inflammatory score induced by HF in liver and renal were reduced by ALP. HF-triggered oxidative stress was inhibited in ALP-treated groups, as evidenced by enhanced SOD1/HO-1/Nrf-2 expressions and reduced thioredoxin-interacting protein (TXNIP)/xanthine oxidase (XO) levels. ALP also suppressed inflammatory response by decreasing pro-inflammatory cytokines through inactivating toll-like receptor 4-nuclear factor kappa B (TLR4-NF-κB) pathway. The anti-oxidant and anti-inflammatory effects of ALP were confirmed in HL-7702 cells. Further, abnormal lipid metabolism caused by HF was alleviated by ALP, which was associated with the decreased Stearoyl-CoA desaturase 1 (SCD1), fatty acid synthase (FAS), sterol element regulatory binding protein 1c (SREBP-1c), Liver X Receptor (LXR)-α, elongases of very long-chain fatty acids (Elovl)-2, p-insulin receptor substrate 1 (IRS1) expressions, and increased PPARα levels. Taken together, the results above indicated that ALP could suppress oxidative stress, reduce inflammatory response and attenuate lipid metabolism, preventing NAFLD.

An in silico approach in predicting the possible mechanism involving restoration of wild-type p53 functions by small molecular weight compounds in tumor cells expressing R273H mutant p53.

R273H mutant p53 is a DNA-contact mutant that renders p53 dysfunctional due to a single substitution of Arg273 for His273. Rescuing R273 mutant p53 implies that a competent molecule would have to bind to the site of DNA-contact hot spots to complement the loss of contact with the DNA-binding domain. Here, curcumin, flavokawain B, and alpinetin were docked against the crystal structure of R273H mutant p53 in silico. Consequently, all the compounds bind to the cavity of R273H mutant p53 with a dissociation constant estimated to have 36.57, 70.77, and 75.11 µM for curcumin, flavokawain B, and alpinetin, respectively. Subsequently, each molecule was able to bind to the R273H mutant p53 by interacting with the DNA-contact hot spot Arg248 and mutant R273H, thereby compensating for the loss of direct contact with the DNA-binding domain. Furthermore, all the molecules were able to induce a direct contact with the consensus site of the DNA binding domain, thus maintaining DNA-contact residues with the DNA. The present findings offer preliminary indirect supporting evidence that small molecular weight compounds may certainly rescue DNA-contact mutant p53, which may lay a foundation for designing a competent and effective molecule capable of rescuing mutant p53 in tumor cells expressing R273H mutant p53.

Spectrometric study of the interaction between Alpinetin and bovine serum albumin using chemometrics approaches

The binding interaction of Alpinetin (APT) with bovine serum albumin (BSA) was studied by fluorescence, UV–visible and synchronous fluorescence spectroscopy (SFS) under simulated physiological conditions. The measured complex spectra were resolved by multivariate curve resolution-alternating least squares (MCR-ALS), yielding a host of data and information, which otherwise would have been impossible to obtain. The extracted profiles corresponded to the spectra of the single species in the APT/BSA mixture. In addition, the presence of the APT–BSA complex was demonstrated, and it was shown that the associated quenching of the fluorescence from the BSA protein resulted from the formation of APT–BSA complex via a static mechanism. The binding constant ( K a(ave) = 2.34 × 10 6 L mol −1) and the number of sites ( n = 1) were obtained by fluorescence methods as were the thermodynamic parameters (Δ H 0, Δ S 0 and Δ G 0). This work suggested that the principal binding between APT to BSA was facilitated by hydrophobic interactions. The thermodynamic parameters for APT were compared to those from the structurally similar Chrysin and Wogonin molecules. It appeared that the entropy parameters were relatively more affected by the small structural changes. SFS from the interaction of BSA and APT showed that the ligand affected the conformation of BSA. The competitive interaction of APT and site makers with BSA indicated site I as the binding area of APT in BSA.

Studies on the constituents of the seeds of Alpinia katsumadai Hayata.

Four new diarylheptanoids [(5R)-trans-1, 7-diphenyl-5-hydroxy-6-hepten-3-one (VIIa), (3S, 5S)-trans-1, 7-diphenyl-3, 5-dihydroxy-1-heptene (VIIIa), trans-1, 7-diphenyl-5-hydroxy-1-heptene (IX) and trans, trans-1, 7-diphenyl-5-hydroxy-4, 6-heptadien-3-one (Xa)] have been isolated from the seeds of Alpinia katsumadai HAYATA, together with trans, trans-farnesol (I), trans-cinnamaldehyde, three flavonoids [alpinetin (II), cardamomin (III) and pinocembrin (IV)], and two known diarylheptanoids [(3S, 5R)-3, 5-dihydroxy-1, 7-diphenylheptane (V) and trans, trans-1, 7-diphenyl-4, 6-heptadien-3-one (VI)], and their structures were determined on the basis of chemical and spectral evidence. This is the first time that trans-cinnamaldehyde, IV, V and VI have been isolated from these seeds.

A note on the constitution of alpinetin

A note on the constitution of alpinetin