Antioxidant, Diabetic and Inflammatory Activities of Alpinia calcarata Roscoe Extract.

BACKGROUND: Alpinia calcarata (AC) Roscoe of Zingiberaceae popularly known as lesser galangal has a widespread occurrence in China, India, Sri-Lanka, Bangladesh, Malaysia, Indonesia and Thailand. Essential oil (Eoil) was obtained from leaves/rhizomes of AC via hydro-distillation process. METHODS: To identify chemical ingredients in oil from leaves/rhizomes of AC through GC/MS technique for volatile components and their anti-oxidant, inflammatory/diabetic activities. RESULTS: The 38 and 65 components were found to make up 99.9 and 99.6 %, respectively in total of Eoil composition of AC leaves/rhizomes. Key chemical constituents were eucalyptol (28.7 % in leaves; 25.4 % in rhizomes), camphor (12.8 % in leaves; 4.2 % in rhizomes), and carotol (9.8 % in leaves; 5.6 % in rhizomes) found in oil of AC leaves/rhizomes. Colorimetric assay showed anti-oxidant activities in leaves and rhizomes are IC50 =71.01±0.71 µg/mL and IC50 =73.83±0.49 µg/mL, respectively in the Eoils. Eoils had high anti-oxidant capabilities in IC50 -values of AC-L-Eoil=43.09±0.82&AC-Rh-Eoil=68.11±0.87 in reducing power in µg/mL was found. Albumin test of rhizome oil had IC50 -values of 15.19±0.25 µg/mL. Concentrations range of 7.81 µg/mL and 250 µg/mL in the Eoils of AC leaves and rhizome, respectively by α-glucosidase inhibition assay. CONCLUSION: Our findings demonstrated that leaf oil was slightly more promising results than rhizome oil of AC extract, which was ultimately showed medicinal potential of secondary metabolites with anti-oxidant, diabetic/inflammatory activities. Further, Eoils of AC have a wide range of pharmacological potential and promising anti-diabetic effects.

Identification of galangin as the bioactive compound from Alpinia calcarata (Haw.) Roscoe rhizomes to inhibit IRAK-1/ MAPK/ NF-κB p65 and JAK-1 signaling in LPS stimulated RAW 264.7 cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Alpinia calcarata (Haw.) Roscoe rhizomes are used to treat diabetes, rheumatism, gastrointestinal problems, inflammatory diseases, cough and respiratory problems in traditional practices. The primary objective of the study is to identify and isolate anti-inflammatory bioactive compounds from A.calcarata rhizomes and to assess its molecular mechanism. MATERIALS AND METHODS: The bioassay-guided fractionation of methanolic extract of A. calcarata rhizomes yielded chloroform fraction as the effective fraction and galangin as the bioactive compound identified by NMR studies. The anti-inflammatory action of galangin was evaluated by determining NO and cytokine production in LPS stimulated RAW264.7 cells. Further, its mechanism was studied on the expression levels of mRNA and protein targets by qPCR and Western blot analysis. RESULTS: Based on the MTT assay, the concentration of 3.1-25 µM of galangin was selected for further studies. Galangin reduced the levels of NO and proinflammatory cytokines (TNF-α, IL-1ß and IL-6) production in LPS induced RAW 264.7 cells in a dose-dependent manner. In addition, the qPCR analysis revealed a reduction in the mRNA expression levels of COX-2, IRAK 1 and JAK 1 in galangin treated LPS stimulated RAW 264.7 cells in a dose-dependent manner. Western blot analysis implicated that galangin has markedly reduced the protein expression levels of cell signaling regulators (JAK-1, IRAK-1, MyD88, MAPK (p38 and ERK) and NF-κB p65). CONCLUSION: From the results, it is evident that the inhibition of these cell signaling regulators has contributed to the anti-inflammatory effects of galangin. To our knowledge, we are the first to report IRAK-1 and JAK-1 as therapeutic targets of galangin for its anti-inflammatory effect.

Investigation of Alpinia calcarata constituent interactions with molecular targets of rheumatoid arthritis: docking, molecular dynamics, and network approach.

Rheumatoid arthritis (RA) is a systemic autoimmune disorder that commonly affects multiple joints of the body. Currently, there is no permanent cure to the disease, but it can be managed with several potent drugs that cause serious side effects on prolonged use. Traditional remedies are considered promising for the treatment of several diseases, particularly chronic conditions, because they have lower side effects compared to synthetic drugs. In folklore, the rhizome of Alpinia calcarata Roscoe (Zingiberaceae) is used as a major ingredient of herbal formulations to treat RA. Phytoconstituents reported in A. calcarata rhizomes are diterpenoids, sesquiterpenoid, flavonoids, phytosterol, and volatile oils. The present study is intended to understand the molecular-level interaction of phytoconstituents present in A. calcarata rhizomes with RA molecular targets using computational approaches. A total of 30 phytoconstituents reported from the plant were used to carry out docking with 36 known targets of RA. Based on the docking results, 4 flavonoids were found to be strongly interacting with the RA targets. Further, molecular dynamics simulation confirmed stable interaction of quercetin with 6 targets (JAK3, SYK, MMP2, TLR8, IRAK1, and JAK1), galangin with 2 targets (IRAK1 and JAK1), and kaempferol (IRAK1) with one target of RA. Moreover, the presence of these three flavonoids was confirmed in the A. calcarata rhizome extract using LC-MS analysis. The computational study suggests that flavonoids present in A. calcarata rhizome may be responsible for RA modulatory activity. Particularly, quercetin and galangin could be potential development candidates for the treatment of RA. Investigation of Alpinia calcarata constituent interactions with molecular targets of rheumatoid arthritis: docking, molecular dynamics, and network approach.

Alpinia calcarata Roscoe: A potential phytopharmacological source of natural medicine.

Alpinia calcarata Roscoe (Family: Zingiberaceae), is a rhizomatous perennial herb, which is commonly used in the traditional medicinal systems in Sri Lanka. Alpinia calcarata is cultivated in tropical countries, including Sri Lanka, India, and Malaysia. Experimentally, rhizomes of Alpinia calcarata are shown to possess antibacterial, antifungal, anthelmintic, antinociceptive, anti-inflammatory, antioxidant, aphrodisiac, gastroprotective, and antidiabetic activities. Phytochemical screening revealed the presence of polyphenols, tannins, flavonoids, steroid glycosides and alkaloids in the extract and essential oil of this plant. Essential oil and extracts from this plant have been found to possess wide range of pharmacological and biological activities. This article provides a comprehensive review of its ethnomedical uses, chemical constituents and the pharmacological profile as a medicinal plant. Particular attention has been given to the pharmacological effects of the essential oil of Alpinia calcarata in this review so that the potential use of this plant either in pharmaceutics or as an agricultural resource can be evaluated.

Amylase production by endophytic fungi Cylindrocephalum sp. isolated from medicinal plant Alpinia calcarata (Haw.) Roscoe

Amylases are among the most important enzymes used in modern biotechnology particularly in the process involving starch hydrolysis. Fungal amylase has large applications in food and pharmaceutical industries. Considering these facts, endophytic fungi isolated from the plant Alpinia calcarata (Haw.) Roscoe were screened for amylolytic activity on glucose yeast extract peptone agar (GYP) medium. Among thirty isolates of endophytic fungi, isolate number seven identified as Cylindrocephalum sp. (Ac-7) showed highest amylolytic activity and was taken for further study. Influence of various physical and chemical factors such as pH, temperature, carbon and nitrogen sources on amylase production in liquid media were studied. The maximal amylase production was found to be at 30ºC and at pH 7.0 of the growth medium. Among the various carbon and nitrogen sources tested, maltose at 1.5% and Sodium nitrate at 0.3% respectively gave optimum amylase production.

Amylase production by endophytic fungi Cylindrocephalumsp. isolated from medicinal plant Alpinia calcarata (Haw.) Roscoe.

Amylases are among the most important enzymes used in modern biotechnology particularly in the process involving starch hydrolysis. Fungal amylase has large applications in food and pharmaceutical industries. Considering these facts, endophytic fungi isolated from the plant Alpinia calcarata (Haw.) Roscoe were screened for amylolytic activity on glucose yeast extract peptone agar (GYP) medium. Among thirty isolates of endophytic fungi, isolate number seven identified as Cylindrocephalum sp. (Ac-7) showed highest amylolytic activity and was taken for further study. Influence of various physical and chemical factors such as pH, temperature, carbon and nitrogen sources on amylase production in liquid media were studied. The maximal amylase production was found to be at 30(º)C and at pH 7.0 of the growth medium. Among the various carbon and nitrogen sources tested, maltose at 1.5% and Sodium nitrate at 0.3% respectively gave optimum amylase production.

Comparative powder microscopy of Alpinia calcarata Roscoe and Alpinia galanga (Linn.) Willd.

Medicinal plant materials are being adulterated in commerce due to many reasons such as similar morphological features, same name as written in classical text, presence of similar active principles in the substituted plant etc., that may badly affect the therapeutic activity of the finished products. Therefore, systematic identification is becoming essential in order to produce standardized finished herbal products. The present study includes two medicinal plant rhizomes; Alpinia calcarata Roscoe (Heenaratta) and A. galanga (Linn.) Willd (Aratta) whose microscopical build up was different from each other and was assessed by standard Pharmacognostical methods. Diagnostic identification characters of A. calcarata were compound starch grins, triangular-shaped starch grains, and plenty of simple starch grains in one parenchyma cell compared to that of A. galanga. Diamond-shaped silica crystals were found only on A. galanga rhizome powder. Present study has revealed an easy technique to identify two similar medicinal plant materials microscopically and this method can also be employed to detect the degree of adulteration in powdered raw medicinal plant materials as well.

The influence of Alpinia calcarata extract on the serum lipid and leptin levels of rats with hyperlipidemia induced by high-fat diet

Objective: To study the effect of ethanolic extract from Alpinia calcarata rhizomes on the serum lipid and leptin levels of rats with hyperlipidemia induced by high-fat diet. Methods: Male wistar rats were divided into six groups: normal, high-fat diet control (HFD) and ethanolic extract of Alpinia calcarata rhizomes (ACRE) (100, 200 and 300 mg/kg; p.o.) was administered to the high fat-diet-induced hyperlipidemic rats for 30 days to evaluate its antihyperlipidemic activity. Atorvastatin (10 mg/kg; p.o.) was used as a standard drug. Results: The results demonstrated that in HFD-induced hyperlipidemic rat, ACRE reduced rat weight gain, triglyceride (TG), total cholesterol levels (TC), low-density lipoprotein (LDL), very low-density lipoprotein (VLDL), total protein (TP) and leptin level (pg/mL). Conclusions: These encouraging findings suggest that ACRE has excellent pharmacological potential to prevent hyperlipidemia.

Safety profile of Alpinia calcarata Roscoe, used in traditional medicine in Sri Lanka / Perfil de seguridad de Alpinia calcarata Roscoe usada en medicina tradicional en Sri Lanka

The aim of the present study was to investigate whether Alpinia calcarata Roscoe (Family: Zingiberaceae) rhizomes have any toxic effects in rats. Wistar rats were used as the experimental model and orally administered hot water extract (HWE) and hot ethanolic extract (HEE) of A. calcarata rhizomes at a dose of 1500 mg/kg respectively for 42 consecutive days. Administration of the HWE or HEE to rats did not result in any chronic toxic effects as evident from their effects on (a) liver function (b) kidney function, (c) hematological parameters such as red blood cell (RBC) count, white blood cell (WBC) count and hemoglobin (Hb) concentration (d) external morphology and wet weights of selected organs. Further, the HWE and the HEE did not appear to mediate any unacceptable effects on food and water intake, percent weight gain, consistency of faeces and color of urine. In conclusion, the results of this study have revealed that the HWE and the HEE of A. calcarata at the doses tested do not produce any serious toxic side effects in rats.

El objetivo del presente estudio fue investigar si los rizomas de Alpinia calcarata Roscoe (Familia: Zingiberaceae) tienen algún efecto tóxico en las ratas. Se utilizaron ratas Wistar como modelo experimental y administrado por vía oral de extracto de agua caliente (EAC) y el extracto de etanol caliente (EEC) de A. rizomas calcarata a una dosis de 1500 mg / kg, respectivamente, durante 42 días consecutivos. La administración de la EAC o EEC a las ratas no produjo ningún efecto crónico tóxico como se desprende de sus efectos sobre la (a) función hepática (b) la función renal, (c) los parámetros hematológicos, como conteo de los glóbulos rojos (GR), conteo de glóbulos blancos celular (GB) y hemoglobina (Hb), (d) morfología externa y el peso húmedo de los órganos seleccionados. Además, el EAC y el EEC no parecen mediar efectos inaceptables en la ingesta de alimentos y agua, porcentaje de aumento de peso, la consistencia de las heces y el color de la orina. En conclusión, los resultados de este estudio han revelado que los extractos EAC y la EEC de A. calcarata en las dosis utilizadas no producen graves efectos secundarios tóxicos en ratas.

Unusual intron in the second exon of a Type III polyketide synthase gene of Alpinia calcarata Rosc.

Plant phenolic compounds form a valuable resource of secondary metabolites having a broad spectrum of biological activities. Type III polyketide synthases play a key role in the formation of basic structural skeleton of the phenolic compounds. As a group of medicinal plants, PKSs with novel features are expected in the genome of Zingiberaceae. The genomic exploration of PKS in Alpinia calcarata conducted in this study identified the presence of an unusual intron at the region forming the second exon of typical PKSs, forming a gateway information of distribution of novel PKSs in Zingiberaceae.

Antioxidant and antifungal activities of essential oil of Alpinia calcarata Roscoe rhizomes.

Antioxidant and antifungal activity were determined for the essential oil of Alpinia calcarata Roscoe (Zingiberaceae) rhizomes. Its antioxidant properties were investigated by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay and thiobarbituric acid reactive substances (TBARS) assay. Butylated hydroxy toluene (BHT) and vitamin E served as positive controls. Antifungal activities were investigated against crop pathogens Curvularia spp. and Colletorichum spp. using the agar plate method. Fifty percent effective concentration (EC(50)) and % antioxidant index of the essential oil were 45 ± 0.4 and 16.1 ± 0.2 for DPPH and TBARS assays, respectively. The degree of, the essential oil's inhibition of the growth of crop pathogens Curvularia spp. and Colletorichum spp. varied with time period its effects were higher than greater than for the positive control, daconil. In conclusion, the essential oil of A. calcarata rhizomes possess moderate antioxidant property and promising antifungal activity.

Standardization of Alpinia calcarata Roscoe rhizomes.

BACKGROUND: Rhizomes of Alpinia calcarata Roscoe (Family: Zingiberaceae) possess several bioactivities and are used in the traditional medicinal systems of Sri Lanka. METHODS: The present investigation was carried out to standardize the rhizomes of A. calcarata by (a) screening for phytochemicals (b) determination of physico-chemical parameters and (c) development of a Densitogram. RESULTS: Phytochemical screening revealed the presence of polyphenols, tannins, flavonoids, steroid glycosides and alkaloids in A. calcarata rhizomes. The percentages of moisture, total ash, acid insoluble ash, water soluble ash, ethanol extractable matter and water extractable matter were of 5.5 - 6.8, 8.3 - 8.8, 0.036 - 0.040, 7.2 - 7.8, 22.6 - 24.8 and 18.6 - 20.5 respectively. CONCLUSION: The results obtained from this study can be used to standardize rhizomes of A. calcarata grown in Sri Lanka.

Unusual intron in the second exon of a Type III polyketide synthase gene of Alpinia calcarata Rosc.

Plant phenolic compounds form a valuable resource of secondary metabolites having a broad spectrum of biological activities. Type III polyketide synthases play a key role in the formation of basic structural skeleton of the phenolic compounds. As a group of medicinal plants, PKSs with novel features are expected in the genome of Zingiberaceae. The genomic exploration of PKS in Alpinia calcarata conducted in this study identified the presence of an unusual intron at the region forming the second exon of typical PKSs, forming a gateway information of distribution of novel PKSs in Zingiberaceae.