Influence of Thermal Treatment on the Composition of Alpinia officinarum Rhizome.

Alpinia officinarum is a representative of the Zingiberaceae family, which is known for its wide use in the food and pharmaceutical industries also due to its precious pharmacological potential. The major aim of the present study was to evaluate the influence of thermal treatment on the composition of the rhizome of Alpinia officinarum and its antioxidant activity. The fresh rhizome was subjected to various thermal treatment processes-boiling, frying and microwave heating during various time intervals-and their composition and antioxidant activity were determined using chromatographic (HPLC - High Performance Liquid Chromatography and HPLC-MS - High Performance Liquid Chromatography Mass Spectrometry) and spectrophotometric (DPPH and TPC - Total Phenolic Content) methods. Pinobanksin was the main compound found in the extract of the fresh rhizome (537.79 mg/kg), followed by galangin (197.7 mg/kg) and zingerone (185.5 mg/kg). The effect of thermal treatment on the rhizome composition was varied. In general, thermal processing significantly decreased the content of active compounds in the rhizome. However, there were some exceptions-boiling for 4 min significantly increased the content of pinobanksin (1162.4 mg/kg) and galangin (280.7 mg/kg), and microwave processing for 4 min increased the content of pinocembrin (213 mg/kg). It was found that boiling and microwave treatment significantly increased the antioxidant activity of the processed rhizomes.

Ultrasonic-Assisted Extraction, Structural Characteristics, and Antioxidant Activities of Polysaccharides from Alpinia officinarum Hance.

Alpinia officinarum Hance, a well known agricultural product in the Lei Zhou peninsula, is generally rich in polysaccharides. In order to enhance the use of A. officinarum Hance polysaccharides (AOP) in functional food, AOP was extracted using an ultrasonic-assisted extraction method, and the ultrasonic extraction parameters of AOP was optimized. Furthermore, this study investigated the physicochemical and antioxidant activities of AOPs. In addition, the structural properties were preliminarily determined using Fourier-transform infrared spectroscopy (FTIR), high performance size exclusion chromatography, and a Zetasizer. Ultimately, this study explored the mechanism underlying the antioxidant activities of AOP. The results showed that the optimal ultrasonic-assisted extraction parameters were as follows: ultrasonic time, 6 min; ratio of water to material, 12 mL/g; and ultrasonic power, 380 W. Under these conditions, the maximum yield of AOPs was 5.72%, indicating that ultrasonic-assisted extraction technology is suitable for extracting AOPs due to the reduced time and water usage. Additionally, AOPs were purified using graded alcohol precipitation, resulting in three fractions (AOP30, AOP50, and AOP70). AOP30 had the lowest molecular weight of 11.07 kDa and mainly consisted of glucose (89.88%). The half inhibitory concentration (IC50) value of AOP30 and AOP70 was lower than that of AOP50 in the ability to scavenge the ABTS radical, while a reverse trend was observed in reducing ferric ions. Notably, the antioxidant activities of AOPs were highly correlated with their polydispersity index (Mw/Mn) and Zeta potential. AOP30, a negatively charged acidic polysaccharide fraction, exhibited electron donating capacities. Additionally, it displayed strong antioxidant abilities through scavenging 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) radicals and reducing ferric ions. In conclusion, the present study suggests that AOP30 could be developed as an antioxidant ingredient for the food industry.

Components study on gastroprotective effect and holistic mechanism of the herbal pair Alpinia officinarum - Cyperus rotundus based on spectrum-effect relationship and integrated transcriptome and metabolome analyses.

ETHNOPHARMACOLOGICAL RELEVANCE: The herbal pair Alpinia officinarum-Cyperus rotundus (HPAC) has an extended history of use in the treatment of gastric ulcers, and its curative effect is definite. AIM OF THE STUDY: To explore the material basis and holistic mechanism of HPAC on ethanol-induced gastric ulcers. MATERIALS AND METHODS: Three chemometrics, GRA, OPLS, and BCA, were used to construct the spectrum-effect relationship between the HPLC fingerprints of HPAC extracts and the bioactivity indices (cell viability; the levels of TNF-α, IL-6, COX-2, and PGE2; and wound healing rate) against GES-1 cell damage to screen the bioactive ingredients. The bioactive components were isolated and validated in vitro. Simultaneously, the effects of HPAC with concentrated bioactive ingredients was tested on ethanol-induced gastric ulcers in vivo, and the mechanism was investigated using transcriptomics and metabolomics. The mechanism was further validated by Western blotting. Finally, the contents of the main components of HPAC were determined before and after compatibility. RESULTS: Twelve bioactive components were screened, and the structures of nine compounds were confirmed. An in vitro verification test showed that DPHA and galangin could protect GES-1 cells from injury, and that their content increased after compatibility. The CH2Cl2 fraction of HPAC (HP-CH2Cl2) can protect mice from ethanol-induced gastric mucosal injury by reducing hemorrhage and decreasing inflammatory cell infiltration. Western blot analysis indicated that this fraction may up-regulate TRPV1 protein and down-regulate PI3K and AKT proteins. CONCLUSIONS: DPHA and galangin may be the bioactive components against ethanol-induced GES-1 cell injury. HP-CH2Cl2 may exert gastroprotective effects by regulating PI3K, AKT and TRPV1 proteins.

Alpinia officinarum Hance extract ameliorates diabetic gastroparesis by regulating SCF/c-kit signaling pathway and rebalancing gut microbiota.

Diabetic gastroparesis (DGP) is a common complication of type 2 diabetes mellitus (T2DM). Alpinia officinarum Hance (AOH) is one of the most commonly used both as a food and folk medicines, which is rich in diarylheptanoids and flavonoids. The gastroprotection and hypoglycemic effect make AOH has great potential in developing of anti-DGP complementary medicine. However, the molecular mechanisms of AOH that act against DGP are yet to be elucidated. In this study, we evaluated the therapeutic effects, the potential molecular mechanism, and the changes of gut microbiota of AOH in DGP. The 5 components of the AOH were analyzed, and the potential signaling pathway of AOH improving DGP was predicted by molecular docking. Subsequently, DGP rat model was constructed using high-fat-irregular-diet, AOH intervention significantly reduced blood glucose levels, increased gastrointestinal propulsion rate, and improved gastric histological morphology in DGP rats. Meanwhile, AOH has been shown to regulate the SCF/c-kit signaling pathway and rebalance the gut microbiota, which may be closely related to its role in improving DGP. Taken together, AOH may play a protective role on DGP through multiple mechanisms, which might pave the road for development and utilization of AOH.

Exploring the Underlying Mechanism of Alpinia officinarum Hance Ameliorating Diabetic Gastroparesis through Combining Network Pharmacology, Molecular Docking, and in Vivo Experimental Verification.

BACKGROUND: Alpinia officinarum Hance (AOH) has a long history in China as a Chinese medicine and exerts the pharmacological effects of antidiabetic and gastrointestinal protection. In traditional Chinese medicine theory, AOH is often combined with other Chinese medicines for the treatment of diabetic gastroparesis (DGP). However, the molecular mechanisms, potential targets, and bioactive ingredients of AOH that act against DGP are yet to be elucidated. In this study, network pharmacology, molecular docking, and experimental study were used to predict the therapeutic effects and the potential molecular mechanism of AOH in DGP. METHODS: Network pharmacology analysis was performed to acquire information on the active chemical ingredients, DGP-related target proteins in AOH, and potential signaling pathway. In addition, molecular docking approach was used to simulate the binding of drugs and targets. Finally, DGP-mice model was used for experimental verification in vivo. Results: Through the network pharmacological research, AKT1 was found to be the core protein in AOH for the treatment of DGP and was mainly involved in the PI3K-AKT signaling pathway. Additionally, the interactions between bioactive compounds and target proteins (PIK3CA and AKT1) were analyzed using molecular docking, which verified the results of network pharmacology. Further in vivo studies indicated that AOH could reduce fasting blood glucose levels, improve gastric emptying rate, and ameliorate biochemical indicators in DGP mice. Moreover, AOH could increase the expressions and phosphorylation levels of PI3K and AKT in the stomach to regulate oxidative stress. CONCLUSIONS: The study has shown that AOH may play a protective role on DGP through mediation of the PI3K-AKT signaling pathway to regulate oxidative stress.

Hepatoprotective effects of Alpinia officinarum rhizome extract on cisplatin-induced hepatotoxicity in rats: A biochemical, histopathological and immunohistochemical study.

BACKGROUND: Alpinia officinarum is a member of the ginger family (Zingiberaceae), which is widely cultivated in Asia and traditionally used for its anti-inflammatory, antimicrobial, and antihyperlipidemic qualities. This study aimed to evaluate the effect of Alpinia officinarum rhizome extract (AORE) on cisplatin (CP)-induced hepatotoxicity in rats. METHODS: Forty-four male rats were divided into six groups including the control group, AORE control group, CP control group, and three groups of CP (7 mg/kg dose, on the 10th day) with AORE (at concentrations of 100, 200 and 400 mg/kg, daily for 14 days). After 14 days, the rats' livers were removed and their liver function was assessed using biochemical marker enzymes including serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) activities and albumin, total protein, and total bilirubin (T. bilirubin). Oxidative stress was assessed by evaluating malondialdehyde concentration and hepatic superoxide dismutase activity, histopathological and immunohistochemical tests were also conducted. RESULTS: Results demonstrated that treatment with AORE reduced the toxicity in levels of the hepatic biomarkers in cp-induced groups. AORE treatment decreased oxidative stress and improved histopathological indexes. Furthermore, immunohistochemical (IHC) investigation showed the B-cell lymphoma 2 (Bcl-2) upsurging and p53 downregulating expression exhibiting the recovery following AORE administration. CONCLUSION: The founding suggested that AORE administration has positive biochemical, histopathological, and immunohistochemical impacts on the ameliorating of hepatotoxicity in CP-induced rats.

Extraction and isolation of diphenylheptanes and flavonoids from Alpinia officinarum Hance using supercritical fluid extraction followed by supercritical fluid chromatography.

In this paper, an off-line combination method of supercritical fluid extraction and supercritical fluid chromatography was developed for the selective extraction and isolation of diphenylheptanes and flavonoids from Alpinia officinarum Hance. The enrichment of target components was successfully achieved using supercritical fluid extraction with the following conditions (8% ethanol as co-solvent at 45°C and 30 MPa for 30 min). Taking full advantage of the complementarity of supercritical fluid chromatography stationary phases, a two-step preparative supercritical fluid chromatography strategy was constructed. The extract was firstly divided into seven fractions on a Diol column (250 × 20 mm internal diameter, 10 µm) within 8 min by gradient elution increasing from 5% to 20% modifier (methanol) at 55 ml/min and 15 MPa. Then the seven fractions were separated by using a 1-AA or a DEA column (250 × 19 mm internal diameter, 5 µm) at 50 ml/min and 13.5 MPa. This two-step strategy showed superior separation ability for structural analogs. As a result, seven compounds, including four diphenylheptanes and three flavonoids with high purity, were successfully obtained. The developed method is also helpful for the extraction and isolation of other structural analogs of traditional Chinese medicines.

Nanographite-assisted matrix solid phase dispersion microextraction of active and toxic compounds from complex food matrices using cyclodextrin aqueous solution as elution solvent.

In this study, a cyclodextrin aqueous solution was used as an environmentally friendly eluent to simultaneously extract active and toxic compounds from food matrices with the aid of nanographite-assisted matrix solid phase dispersion microextraction (NG-MSPDM). The NG-MSPDM procedure was optimized by single-factor experiments and response surface methodology to obtain optimum conditions. The proposed method achieved excellent linearity at 0.10-20 µg/mL for all target analytes with a coefficient of correction (R2) ≥ 0.9909, limits of detection < 52.01 ng/mL, satisfactory reproducibility below 3.21 %, and acceptable recoveries of 82.0-112 %. To accurately determine the target components in the complex matrix, collision cross-section values of the analytes were obtained using ion mobility quadrupole time-of-flight mass spectrometry (IM-Q-TOF/MS). Results indicated that the NG-MSPDM method successfully achieved the simultaneous extraction of flavonoids and phenoxyacetic herbicides from Alpinia officinarum.

The ameliorative effects of Alpinia officinarum rhizome hydroalcoholic extract on cisplatin-induced testicular toxicity in rats.

OBJECTIVE: In this study we evaluated the influence of Alpinia officinarum rhizome extract (AO) on the alleviation of testicular damage induced by cisplatin in rats. METHODS: The study groups included the control group, AO-administered group, cisplatin-administered group, and three groups administered with cisplatin and AO (different concentrations of 100, 200, and 400 mg/kg). On the 14th day we removed the testes of the rats, and the testicular organ parameters were measured. Moreover, through the malondialdehyde concentration we assessed the oxidative stress and superoxide dismutase (SOD) activity of the testes and ran a histopathological analysis. RESULTS: The results demonstrated that cisplatin-induced oxidative stress and severe testicular damage on the AO-administered group showed no harm compared with the control group. AO- treatment in cisplatin-received rats led to the reduction of oxidative stress, enhancement of SOD activity, and prevention of testicular damage. The lowest testis damage was attributed to the group which received 400 mg/kg of AO compared to 100 and 200 mg/kg. CONCLUSIONS: Overall, the Cis+/AO+400 group had the best antioxidant effect. The findings could lead to changes in cancer care guidelines that incorporate phytochemicals, making cancer therapies safer.

Alpinia officinarum mediated copper oxide nanoparticles: synthesis and its antifungal activity against Colletotrichum gloeosporioides.

Green synthesis offers an environmentally friendly and cost-effective alternative for the synthesis of copper oxide nanoparticles (CuO NPs). In this study, the synthesis of CuO NPs was optimized by using copper sulfate (CuSO4) and the aqueous extract of Alpinia officinarum and its antifungal activity were investigated. The synthesized CuO NPs were characterized by UV-visible spectroscopy (UV-vis), X-ray diffraction (XRD), Fourier-transform infrared radiation spectroscopy (FT-IR), scanning electron microscope (SEM), energy dispersive spectroscopy (EDS), dynamic light scattering (DLS), and transmission electron microscopy (TEM). The results showed that the optimized conditions for the synthesis of CuO NPs were 1:2 ratio of extract and CuSO4 solution, pH 7, and 30 °C. The characteristic UV-vis peak of A. officinarum synthesized CuO NPs was at 264 nm. The synthesized CuO NPs had high crystallinity and purity and were spherical in morphology with the mean size of 46.40 nm. The synthesized CuO NPs reduced the fungal growth of Colletotrichum gloeosporioides in a dose-dependent manner. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of the CuO NPs were 125 µg·mL-1 and 500 µg·mL-1, respectively. The antifungal activity of CuO NPs may be attributed to its ability to deform the structure of fungal hyphae, induce excessive reactive oxygen species accumulation and lipid peroxidation in fungi, disrupt the mycelium cell membrane, and result cellular leakage.

Alpinia officinarum Rhizome ameliorates the UVB induced photoaging through attenuating the phosphorylation of AKT and ERK.

BACKGROUND: Chronic ultraviolet (UV) exposure is one of the major external factors in skin aging, and repetitive UVB exposure induces extracellular matrix (ECM) damage as well as metabolic disease. Alpinia officinarum Rhizome (AOR) is a medicinal plant that has been traditionally used for treating rheumatism and whooping cough. However, the antiphotoaging effects of AOR remain unclear. We investigated the protective effects of water extracts of AOR (WEAOR) in terms of UVB-mediated ECM damage, wrinkle formation, inflammatory responses, and intracellular signaling on hairless mice and NIH-3T3 skin fibroblast cells. METHODS: WEAOR was administered to UVB-irradiated hairless mice. Wrinkle formation was assessed using the replica assay, epidermal changes through H&E staining, and collagen contents in mice skin through Masson's trichrome staining. The expression of procollagen type-1 (COL1A1), metalloproteinase-1a (MMP-1a), and inflammatory cytokines (IL-6, IL-8, and MCP-3) in hairless mice skin and NIH-3T3 cells was investigated through qRT-PCR. The effects of WEAOR or signaling inhibitors on UVB-induced expression of intracellular mitogen-activated protein kinases (MAPKs) were estimated by Western blotting and qRT-PCR, respectively. RESULTS: Topical WEAOR significantly attenuated the UVB-induced wrinkle formation and epidermal thickening in the skin of hairless mice. WEAOR treatment also attenuated the UVB-induced expression of MMP-1a and COL1A1 and recovered the reduction of collagen content in mouse skin. These effects were confirmed in NIH-3T3 skin fibroblast cells. WEAOR treatment restored the UVB-induced COL1A1 and MMP-1a gene expression and attenuated the UVB-induced expression of IL-6, IL-8, and MCP-3 in NIH-3T3 cells. Notably, WEAOR attenuated UVB-induced phosphorylation of AKT and ERK, but not that of p38 and JNK in NIH-3T3 cells. In addition, the administration of AKT and ERK inhibitors restored the UVB-induced expression of MMP-1a and COL1A1 to an equal extent as WEAOR in NIH-3T3 cells. CONCLUSIONS: The antiphotoaging properties of WEAOR were first evaluated in this study. Our results suggest that WEAOR may be a potential antiphotoaging agent that ameliorates UVB-induced photoaging processes via the AKT and ERK signaling pathways.

DPHB, a diarylheptane from Alpinia officinarum Hance, ameliorates insulin resistance: A network pharmacology and in vitro study.

(4E)-7-(4-Hydroxy-3-methoxyphenyl)-1-phenylhept-4-en-3-one (DPHB) derived from A. officinarum Hance has been reported to exert anti-inflammatory and anti-insulin resistance (IR) effects. We explored the molecular mechanism of DPHB ameliorating IR through network pharmacological prediction and in vitro analysis. The PI3K/AKT and TNF signaling pathways are the core pathways for DPHB to exert anti-IR, and the key proteins of this pathway were confirmed by molecular docking. In the IR-3T3-L1 adipocyte model, DPHB significantly promoted glucose uptake and the glucose transporter type 4 (GLUT4) translocation. In addition, DPHB significantly improved lipid accumulation, triglyceride content, and the mRNA expression of key adipokines [such as peroxisome proliferator-activated receptors-gamma (PPARγ), CCAAT enhancer-binding protein alpha (C/EBPα), and sterol regulatory element-binding protein-1 (SREBP-1)]. DPHB inhibited the protein expression of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and phosphorylated nuclear factor-κB (NF-kB), as well as promoted the expression of phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), phosphorylated PI3K, and phosphorylated AKT. More interestingly, validation of the PI3K inhibitor LY294002 revealed that these changes were dependent on the activation of PI3K. Our cumulative findings thereby validate the potential of DPHB to alleviate and treat IR and the related diseases by regulating the PI3K/AKT and TNF-α signaling pathways.

Synthesis of silver nanoparticles using Alpinia officinarum rhizome extract induces apoptosis through down-regulating Bcl-2 in human cancer cells.

In this study, silver nanoparticles were synthesized using Alpinia officinarum rhizome extract via an eco-friendly green synthesis method. The silver nanoparticles (AO-AgNPs) were characterized by UV-Vis spectrometry, scanning electron microscopy, energy-dispersive X-ray spectroscopy, and dynamic light scattering. Further, the cytotoxic and apoptotic effects of AO-AgNPs were investigated in human cancer cells with different tissue origins via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and flow cytometric analyses, respectively. The expression levels of anti-apoptotic Bcl-2 protein were evaluated via a real-time polymerase chain reaction. The synthesized AO-AgNPs induced a significant cytotoxic effect in all tested cancer cells but not in normal cells. AO-AgNPs induced the percentage of apoptotic cells and reduced the levels of anti-apoptotic Bcl-2 mRNA levels in cancer cells. These results demonstrate the potential application of AO-AgNPs in cancer treatment.

Antibacterial Activity and Components of the Methanol-Phase Extract from Rhizomes of Pharmacophagous Plant Alpinia officinarum Hance.

The rhizomes of Alpinia officinarum Hance (known as the smaller galangal) have been used as a traditional medicine for over 1000 years. Nevertheless, little research is available on the bacteriostatic activity of the herb rhizomes. In this study, we employed, for the first time, a chloroform and methanol extraction method to investigate the antibacterial activity and components of the rhizomes of A. officinarum Hance. The results showed that the growth of five species of pathogenic bacteria was significantly inhibited by the galangal methanol-phase extract (GMPE) (p < 0.05). The GMPE treatment changed the bacterial cell surface hydrophobicity, membrane fluidity and/or permeability. Comparative transcriptomic analyses revealed approximately eleven and ten significantly altered metabolic pathways in representative Gram-positive Staphylococcus aureus and Gram-negative Enterobacter sakazakii pathogens, respectively (p < 0.05), demonstrating different antibacterial action modes. The GMPE was separated further using a preparative high-performance liquid chromatography (Prep-HPLC) technique, and approximately 46 and 45 different compounds in two major component fractions (Fractions 1 and 4, respectively) were identified using ultra-HPLC combined with mass spectrometry (UHPLC-MS) techniques. o-Methoxy cinnamaldehyde (40.12%) and p-octopamine (62.64%) were the most abundant compounds in Fractions 1 and 4, respectively. The results of this study provide data for developing natural products from galangal rhizomes against common pathogenic bacteria.

Unraveling inhibitory effects of Alpinia officinarum Hance and curcumin on methylimidazole and acrylamide in cookies and possible pathways revealed by electron paramagnetic resonance.

The synchronous mitigative effects of Alpinia officinarum Hance (AOH) and curcumin on the generation of methylimidazole and acrylamide in cookies were investigated. Possible mechanisms related to quenching free radicals, reducing lipid oxidation and eliminating carbonyl intermediates were explored by electron paramagnetic resonance (EPR) and HPLC. The total methylimidazole and acrylamide contents raised with an increase in heating temperature and time, and reached a maximum at 200 °C for 11 min. AOH and curcumin reduced methylimidazole and acrylamide simultaneously; the maximum inhibition rates for methylimidazole and acrylamide were 51.55% (0.015% curcumin) and 73.66% (1.5% AOH). Alkyl free radicals and HO· were proved to be the critical free radicals for methylimidazole and acrylamide, AOH and curcumin quenched these radicals in a dose-dependent manner. The lipid oxidation, active carbonyl intermediates glyoxal, methylglyoxal, and acrylaldehyde were also reduced by AOH and curcumin simultaneously, which may be resulted from the quenching of free radicals.

Vasodilatory Effect of Alpinia officinarum Extract in Rat Mesenteric Arteries.

BACKGROUND: Alpinia officinarum (A. officinarum) is known to exhibit a beneficial effect for anti-inflammatory, anti-oxidant, and anti-hyperlipidemic effects. However, no sufficient research data are available on the cardiovascular effect of A. officinarum. Thus, in this study, we investigate whether A. officinarum extract has direct effects on vascular reactivity. METHODS: To examine whether A. officinarum extract affects vascular functionality, we measured isometric tension in rat mesenteric resistance arteries using a wire myograph. After arteries were pre-contracted with high-K+ (70 mM), phenylephrine (5 µM), or U46619 (1 µM), A. officinarum extract was treated. RESULTS: A. officinarum extract induced vasodilation in a concentration-dependent manner, and this effect was endothelium independent. To further investigate the mechanism, we incubated arteries in a Ca2+-free and high-K+ solution, followed by the cumulative addition of CaCl2 (0.01-2.5 mM) with or without A. officinarum extract (30 µg/mL). Pre-treatment of A. officinarum extract reduced the contractile responses induced by cumulative administration of Ca2+, which suggests that extracellular Ca2+ influx was inhibited by the treatment of A. officinarum extract. These results were associated with a reduction in phosphorylated MLC20 in VSMCs treated with A. officinarum extract. Furthermore, eucalyptol, an active compound of A. officinarum extract, had a similar effect as A. officinarum extract, which causes vasodilation in mesenteric resistance arteries. CONCLUSION: A. officinarum extract and its active compound eucalyptol induce concentration-dependent vasodilation in mesenteric resistance arteries. These results suggest that administration of A. officinarum extract could exert beneficial effects to treat high blood pressure.

A diarylheptanoid compound from Alpinia officinarum Hance ameliorates high glucose-induced insulin resistance by regulating PI3K/AKT-Nrf2-GSK3ß signaling pathways in HepG2 cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Alpinia officinarum Hance, a perennial natural medicine-food herb, has been traditionally used to treat colds, stomachache, and diabetes for thousands of years. 1,7-Diphenyl-4E-en-3-heptanone (DPH5), a diarylheptanoid isolated from the rhizome of A. officinarum has been reported to be safe and to have antioxidant and hypoglycemic effects, suggesting its potential in the treatment of insulin resistance (IR). AIM OF THE STUDY: Aim of to investigate the protective effect of DPH5 on IR and elucidate its underlying mechanism of action. MATERIALS AND METHODS: HepG2 cells were used as the research objects. Glucose uptake and reactive oxygen species (ROS) levels in high glucose-induced insulin-resistant HepG2 cells were assessed using flow cytometry. Glucose consumption and the levels of malondialdehyde (MDA) and superoxide dismutase (SOD) were analyzed using the corresponding assay kits. The expression of mRNA and proteins related to insulin signaling, glucose metabolism, and antioxidant factor, including insulin receptor substrate-1 (IRS1), phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), translocation of glucose transporter-4, glycogen synthase kinase-3ß (GSK3ß), glucokinase (GCK), pyruvate kinase (PK), phosphoenolpyruvate carboxykinase (PEPCK), glucose-6-phosphatase (G6Pase), nuclear factor-erythroid 2 related factor 2 (Nrf2), heme oxygenase-1 (HO-1), NADPH quinoneoxidoreductase (NQO1), and glutathione peroxidase (GSH-Px) was determined using real-time quantitative polymerase chain reaction and western blotting. Furthermore, molecular docking was performed to determine the spatial mechanism of DPH5 on the key targets PI3K, AKT, Nrf2, and GSK3ß. RESULTS: DPH5 could improve IR that manifested as increased glucose uptake and glucose consumption in insulin-resistant HepG2 cells. Moreover, DPH5 could enhance antioxidant capacity by activating Nrf2/HO-1 elements, including increasing Nrf2, HO-1, SOD, NQO1, and GSH-Px expression and reducing MDA, ROS, and JNK levels, thereby improving oxidative stress and ultimately alleviating IR. Additionally, DPH5 could promote the expression of IRS1, PI3K, AKT, GSK3ß, GCK, and PK, and downregulate the expression of PEPCK and G6pase, thereby accelerating glucose utilization and enhancing insulin sensitivity. The mechanism underlying the effect of DPH5 in alleviating IR was related to the PI3K/AKT- and Nrf2/HO-1-mediated regulation of the GSK3ß signaling pathway, and the results were further confirmed using the specific inhibitors LY294002 and ML385. Results from molecular docking indicated that there were different regulatory sites and interacting forces between DPH5 and PI3K, AKT, Nrf2, and GSK3ß; however, the binding force was relatively strong. CONCLUSIONS: DPH5 improved oxidative stress and glucose metabolism via modulating the PI3K/AKT-Nrf2-GSK3ß pathway, thereby ameliorating IR. Overall, our findings suggest the potential of DPH5 as a natural medicine to treat type-2 diabetes mellitus.

Hypoglycemic, hypolipidemic and hepatoprotective effects of Alpinia officinarum on nicotinamide/streptozotocin induced type II diabetic rats.

OBJECTIVES: Alpinia officinarum Hance, commonly known as lesser galangal, is a member of the ginger family (Zingiberaceae) traditionally used for many decades to treat inflammation, pain, stomach ache and cold. In the present study, the antidiabetic and hypolipidemic potentials of the hydroalcoholic extract of A. officinarum (AO) were investigated in the nicotinamide/streptozotocin induced type II diabetic rats. METHODS: Male Wistar rats were divided into following six groups: Group I was normal control rats. Group II: normal diabetic control, Group III: Diabetic rats treated with glibenclamide (0.25 mg/kg), IV, V and VI: Diabetic rats treated with 100, 200 and 500 mg/kg AO hydroalcoholic extract by daily gavage for 28 days, respectively. At the end of treatment, biochemical analysis, histological study, phytochemical analysis and acute toxicity tests were carried out. RESULTS: The results show significant reduction in blood glucose, serum lipid profiles, and liver enzyme levels in diabetic rats compared with diabetic control in AO treated group. CONCLUSIONS: In conclusion, the present study demonstrated that AO extract had significant (p<0.05) antidiabetic and anti-hyperlipidemia effects in addition to hepatoprotective effect in type II diabetic rats.

Comparison for quantification of eight components in Alpinia officinarum Hance by using high-performance liquid chromatography coupled with diode array detector and charged aerosol detector with individual and substitute reference compound.

An efficient HPLC-DAD-CAD method was developed and compared for simultaneous quantification of four flavonoids and four diarylheptanoids in Alpinia officinarum Hance (A. officinarum) using individual and substitute reference compound. All calibration curves for investigated analytes showed good linear regression (R2> 0.9991). The LODs of investigated compounds for DAD and CAD were 0.15-7.92 ng (0.03-1.58 µg/mL) and 2.91-3.95 ng (0.58-0.79 µg/mL), respectively, whereas the LOQs were 0.52-26.39 ng (0.10-5.28 µg/mL) for DAD, and 9.70-13.18 ng (1.94-2.64 µg/mL) for CAD. Recoveries of all analytes, which ranged from 96.58% to 100.06% for DAD, and from 96.29% to 99.61% for CAD, were acceptable. According to the quantitative results, the eight compounds in A. officinarum can be accurately quantified with individual calibration curves by two detectors. In addition, to overcome the bottleneck of shortage of reference standards, diphenylheptane A and galangin, respectively, were selected for direct or calibrated quantitative determination of other diarylheptanoids and flavonoids in A. officinarum. The results showed the contents of eight components in A. officinarum determined by these methods were similar, which suggested that substitute reference compound was suitable for quantification of its analogues.