海南蒲桃提取物对L6细胞砷中毒缓解作用的体外实验(英文)
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摘要
目的:研究海南蒲桃提取物对砷酸盐引起的L6骨骼肌细胞葡萄糖内稳态破坏及线粒体功能异常的缓解作用。方法:通过测量多个指标如丙酮酸激酶活性、葡糖激酶、线粒体膜电位等衡量细胞内葡萄糖水平及线粒体功能,并测量相关标志物的蛋白质及mRNA表达情况,如葡萄糖转运体4、胰岛素受体基质1、胰岛素受体基质2、葡糖激酶等以分析可能有关的信号通路。结果:海南蒲桃提取物能够通过葡萄糖转运体4通路改善砷中毒的L6细胞内诸多标志物的表达,使其正常化,与模型组相比差异有统计学意义。结论:海南蒲桃提取物对细胞砷中毒有明显的缓解作用,未来可考虑将其用于治疗砷中毒相关疾病如高血糖症等。
OBJECTIVE: To determine the ameliorative potentials of Syzygium jambolanum (SJ) extract in L6 skeletal muscle cells in regard to arsenic-induced impairment of optimum glucose homeostasis and improper functioning of mitochondria. METHODS: Several study parameters like glucose level and mitochondrial functioning through indexes of pyruvate kinase, glucokinase and mitochondrial membrane potential were assessed. The expression of the relevant marker proteins and mRNAs like glucose transporter 4 (GLUT4), insulin receptor substrate 1 (IRS1), IRS2 and glucokinase for tracking down the signalling cascade were critically analyzed. RESULTS: Introduction of SJ extract could bring about positive modulation of various markers, by acting on GLUT4, thereby bringing about an attenuation of the arsenite-induced toxic conditions in L6 cells. CONCLUSION: Syzygium jambolanum extract has considerable ameliorating potentials against arsenic-induced glucose imbalance and stress and has possibility of therapeutic use in the management of arsenic-induced toxicity including hyperglycemia.
OBJECTIVE: To determine the ameliorative potentials of Syzygium jambolanum (SJ) extract in L6 skeletal muscle cells in regard to arsenic-induced impairment of optimum glucose homeostasis and improper functioning of mitochondria. METHODS: Several study parameters like glucose level and mitochondrial functioning through indexes of pyruvate kinase, glucokinase and mitochondrial membrane potential were assessed. The expression of the relevant marker proteins and mRNAs like glucose transporter 4 (GLUT4), insulin receptor substrate 1 (IRS1), IRS2 and glucokinase for tracking down the signalling cascade were critically analyzed. RESULTS: Introduction of SJ extract could bring about positive modulation of various markers, by acting on GLUT4, thereby bringing about an attenuation of the arsenite-induced toxic conditions in L6 cells. CONCLUSION: Syzygium jambolanum extract has considerable ameliorating potentials against arsenic-induced glucose imbalance and stress and has possibility of therapeutic use in the management of arsenic-induced toxicity including hyperglycemia.
引文
1 Jomova K,Jenisova Z,Feszterova M,Baros S,LiskaJ,Hudecova D,Rhodes CJ,Valko M.Arsenic:toxicity,oxidative stress and human disease.J Appl Toxicol.2011;31(2):95-107.
2 Vigo JB,Ellzey JT.Effects of arsenic toxicity at thecellular level:a review.Tex J Micros.2006;37(2):45-49.
3 Longnecker MP,Daniels JL.Environmental contaminantsas etiologic factors for diabetes.Environ Health Perspect.2001;109(Suppl 6):871-876.
4 Frde TS,Medeiros YS.Animal models to test drugswith potential antidiabetic activity.J Ethnopharmacol.2008;115(2):173-183.
5 Merry TL,McConell GK.Skeletal muscle glucose uptakeduring exercise:a focus on reactive oxygen species andnitric oxide signaling.IUBMB Life.2009;61(5):479-484.
6 Rajasekar V,Kirubanandan S,Lakshmi BS.Assessmentof anti-diabetic of Syzygium jambolanumusing in vitromodel.Am J Infect Dis.2009;5(2):99-105.
7 McDowell HE,Eyers PA,Hundal HS.Regulation ofSystem A amino acid transport in L6rat skeletal musclecells by insulin,chemical and hyperthermic stress.FEBS Lett.1998;441(1):15-19.
8 Gohil T,Pathak N,Jivani N,Devmurari V,Patel J.Treatment with extracts of Eugenia jambolanaseed andAegle marmelos leaf extracts prevents hyperglycemiaand hyperlipidemia in alloxan induced diabetic rats.AfrJ Pharm Pharm.2010;4(5):270-275.
9 Grover JK,Vats V,Rathi SS.Anti-hyperglycemiceffect of Eugenia jambolana and Tinospora cordifoliain experimental diabetes and their effects on key metabolicenzymes involved in carbohydrate metabolism.J Ethno-pharmacol.2000;73(3):461-470.
10 Samadder A,Chakraborty D,De A,Bhattacharyya SS,Bhadra K,Khuda-Bukhsh AR.Possible signaling cascadesinvolved in attenuation of alloxan-induced oxidativestress and hyperglycemia in mice by ethanolic extract ofSyzygium jambolanum:drug-DNA interaction withcalf thymus DNA as target.Eur J Phar Sci.2011;44(3):207-217.
11 Bhattacharyya SS,Paul S,De A,Das D,Samadder A,Boujedaini N,Khuda-Bukhsh AR.Poly(lactide-co-glycolide)acid nanoencapsulation of a synthetic coumarin:cytotoxicityand bio-distribution in mice,in cancer cell line andinteraction with calf thymus DNA as target.ToxicolAppl Pharmacol.2011;253(3):270-281.
12 Mosmann T.Rapid colorimetric assay for cellular growth and survival:application to proliferation and cytotoxicityassays.J Immunol Methods.1983;65(1-2):55-63.
13 Jung KH,Ha E,Kim MJ,Uhm YK,Kim HK,HongSJ,Chung JH,Yim SV.Ganoderma lucidum extractstimulates glucose uptake in L6 rat skeletal musclecells.Acta Biochim Pol.2006;53(3):597-601.
14 Mokrasch LC.Analysis of hexose phosphates and sugarmixtures with the anthrone reagent.J Biol Chem.1954;208(1):55-59.
15 Mazurek S,Grimm H,Boschek CB,Vaupel P,EigenbrodtE.Pyruvate kinase type M2:a crossroad in the tumormetabolome.Br J Nutr.2002;87(Suppl 1):S23-S29.
16 Sedlak J,Lindsay RH.Estimation of total,protein-bound,and nonprotein sulfhydryl groups in tissue with Ellman'sreagent.Anal Biochem.1968;25(1):192-205.
17 Ratha J,Majumdar KN,Mandal SK,Bera R,Sarkar C,Saha B,Mandal C,Saha KD,Bhadra R.A sphingolipidrich lipid fraction isolated from attenuated Leishmaniadonovani promastigote induces apoptosis in mouse andhuman melanoma cells in vitro.Mol Cell Biochem.2006;290(1-2):113-123.
18 Dhiman R,Kathania M,Raje M,Majumdar S.Inhibitioof bfl-1/A1by siRNA inhibits mycobacterial growth iTHP-1 cells by enhancing phagosomal acidificationBiochim Biophys Acta.2008;1780(4):733-742.
19 Biswas R,Mandal SK,Dutta S,Bhattacharyya SS,Boujedaini N,Khuda-Bukhsh AR.Thujone-rich fraction ofThuja occidentalis demonstrates major anti-cancer potentials:evidences from in vitro studies on A375cells.EvidBased Complement Alternat Med.2011;2011:568148.
20 Bhattacharyya SS,Paul S,Khuda-Bukhsh AR.Encapsulatedplant extract(Gelsemium sempervirens )poly (lactide-co-glycolide)nanoparticles enhance cellular uptake andincrease bioactivity in vitro.Exp Biol Med.2010;235(6):678-688.
21 Paul S,Bhattacharyya SS,Samaddar A,Boujedaini N,Khuda-Bukhsh AR.Anticancer potentials of root extract ofPolygala senegaagainst benzo[a]pyrene-induced lungcancer in mice.J Chin Integr Med.2011;9(3):320-327.English with abstract in Chinese.Paul S,Bhattacharyya SS,Samaddar A,Boujedaini N,Khuda-Bukhsh AR.美洲远志根提取物对苯并芘致肺癌小鼠的抗肿瘤作用.中西医结合学报.2011;9(3):320-327.
22 Zierler K,Rogus EM.Hyperpolarization as a mediatorof insulin action:increased muscle glucose uptake inducedelectrically.Am J Physiol.1980;239(1):113-123.
23 Klip A,Ramlal T,Walker D.Insulin stimulation ofglucose uptake and the transmembrane potential of musclecells in culture.FEBS Lett.1986;205(1):11-14.
24 Maechler P,Wollheim CB.Mitochondrial function in normal and diabeticβ-cells.Nature.2001;414(6865):807-812.
25 Massague J,Pilch PF,Czech MP.Electrophoreticresolution of three major insulin receptor structureswith unique subunit stoichiometries.Proc Natl Acad SciU S A.1980;77(12):7137-7141.
26 Rosen BP.Biochemistry of arsenic detoxification.FEBSLett.2002;529(1):86-92.
27 Delnomdedieu M,Basti MM,Styblo M,Otvos JD,Thomas DJ.Complexation of arsenic species in rabbiterythrocytes.Chem Res Toxicol.1994;7(5):621-627.
2 Vigo JB,Ellzey JT.Effects of arsenic toxicity at thecellular level:a review.Tex J Micros.2006;37(2):45-49.
3 Longnecker MP,Daniels JL.Environmental contaminantsas etiologic factors for diabetes.Environ Health Perspect.2001;109(Suppl 6):871-876.
4 Frde TS,Medeiros YS.Animal models to test drugswith potential antidiabetic activity.J Ethnopharmacol.2008;115(2):173-183.
5 Merry TL,McConell GK.Skeletal muscle glucose uptakeduring exercise:a focus on reactive oxygen species andnitric oxide signaling.IUBMB Life.2009;61(5):479-484.
6 Rajasekar V,Kirubanandan S,Lakshmi BS.Assessmentof anti-diabetic of Syzygium jambolanumusing in vitromodel.Am J Infect Dis.2009;5(2):99-105.
7 McDowell HE,Eyers PA,Hundal HS.Regulation ofSystem A amino acid transport in L6rat skeletal musclecells by insulin,chemical and hyperthermic stress.FEBS Lett.1998;441(1):15-19.
8 Gohil T,Pathak N,Jivani N,Devmurari V,Patel J.Treatment with extracts of Eugenia jambolanaseed andAegle marmelos leaf extracts prevents hyperglycemiaand hyperlipidemia in alloxan induced diabetic rats.AfrJ Pharm Pharm.2010;4(5):270-275.
9 Grover JK,Vats V,Rathi SS.Anti-hyperglycemiceffect of Eugenia jambolana and Tinospora cordifoliain experimental diabetes and their effects on key metabolicenzymes involved in carbohydrate metabolism.J Ethno-pharmacol.2000;73(3):461-470.
10 Samadder A,Chakraborty D,De A,Bhattacharyya SS,Bhadra K,Khuda-Bukhsh AR.Possible signaling cascadesinvolved in attenuation of alloxan-induced oxidativestress and hyperglycemia in mice by ethanolic extract ofSyzygium jambolanum:drug-DNA interaction withcalf thymus DNA as target.Eur J Phar Sci.2011;44(3):207-217.
11 Bhattacharyya SS,Paul S,De A,Das D,Samadder A,Boujedaini N,Khuda-Bukhsh AR.Poly(lactide-co-glycolide)acid nanoencapsulation of a synthetic coumarin:cytotoxicityand bio-distribution in mice,in cancer cell line andinteraction with calf thymus DNA as target.ToxicolAppl Pharmacol.2011;253(3):270-281.
12 Mosmann T.Rapid colorimetric assay for cellular growth and survival:application to proliferation and cytotoxicityassays.J Immunol Methods.1983;65(1-2):55-63.
13 Jung KH,Ha E,Kim MJ,Uhm YK,Kim HK,HongSJ,Chung JH,Yim SV.Ganoderma lucidum extractstimulates glucose uptake in L6 rat skeletal musclecells.Acta Biochim Pol.2006;53(3):597-601.
14 Mokrasch LC.Analysis of hexose phosphates and sugarmixtures with the anthrone reagent.J Biol Chem.1954;208(1):55-59.
15 Mazurek S,Grimm H,Boschek CB,Vaupel P,EigenbrodtE.Pyruvate kinase type M2:a crossroad in the tumormetabolome.Br J Nutr.2002;87(Suppl 1):S23-S29.
16 Sedlak J,Lindsay RH.Estimation of total,protein-bound,and nonprotein sulfhydryl groups in tissue with Ellman'sreagent.Anal Biochem.1968;25(1):192-205.
17 Ratha J,Majumdar KN,Mandal SK,Bera R,Sarkar C,Saha B,Mandal C,Saha KD,Bhadra R.A sphingolipidrich lipid fraction isolated from attenuated Leishmaniadonovani promastigote induces apoptosis in mouse andhuman melanoma cells in vitro.Mol Cell Biochem.2006;290(1-2):113-123.
18 Dhiman R,Kathania M,Raje M,Majumdar S.Inhibitioof bfl-1/A1by siRNA inhibits mycobacterial growth iTHP-1 cells by enhancing phagosomal acidificationBiochim Biophys Acta.2008;1780(4):733-742.
19 Biswas R,Mandal SK,Dutta S,Bhattacharyya SS,Boujedaini N,Khuda-Bukhsh AR.Thujone-rich fraction ofThuja occidentalis demonstrates major anti-cancer potentials:evidences from in vitro studies on A375cells.EvidBased Complement Alternat Med.2011;2011:568148.
20 Bhattacharyya SS,Paul S,Khuda-Bukhsh AR.Encapsulatedplant extract(Gelsemium sempervirens )poly (lactide-co-glycolide)nanoparticles enhance cellular uptake andincrease bioactivity in vitro.Exp Biol Med.2010;235(6):678-688.
21 Paul S,Bhattacharyya SS,Samaddar A,Boujedaini N,Khuda-Bukhsh AR.Anticancer potentials of root extract ofPolygala senegaagainst benzo[a]pyrene-induced lungcancer in mice.J Chin Integr Med.2011;9(3):320-327.English with abstract in Chinese.Paul S,Bhattacharyya SS,Samaddar A,Boujedaini N,Khuda-Bukhsh AR.美洲远志根提取物对苯并芘致肺癌小鼠的抗肿瘤作用.中西医结合学报.2011;9(3):320-327.
22 Zierler K,Rogus EM.Hyperpolarization as a mediatorof insulin action:increased muscle glucose uptake inducedelectrically.Am J Physiol.1980;239(1):113-123.
23 Klip A,Ramlal T,Walker D.Insulin stimulation ofglucose uptake and the transmembrane potential of musclecells in culture.FEBS Lett.1986;205(1):11-14.
24 Maechler P,Wollheim CB.Mitochondrial function in normal and diabeticβ-cells.Nature.2001;414(6865):807-812.
25 Massague J,Pilch PF,Czech MP.Electrophoreticresolution of three major insulin receptor structureswith unique subunit stoichiometries.Proc Natl Acad SciU S A.1980;77(12):7137-7141.
26 Rosen BP.Biochemistry of arsenic detoxification.FEBSLett.2002;529(1):86-92.
27 Delnomdedieu M,Basti MM,Styblo M,Otvos JD,Thomas DJ.Complexation of arsenic species in rabbiterythrocytes.Chem Res Toxicol.1994;7(5):621-627.