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紫玉米须:一种潜在的抗肥胖剂,可抑制脂肪生成,并诱导脂肪细胞的脂解和凋亡。

Purple corn silk: A potential anti-obesity agent with inhibition on adipogenesis and induction on lipolysis and apoptosis in adipocytes.

作者信息

Chaiittianan Rungsiri, Sutthanut Khaetthareeya, Rattanathongkom Ariya

机构信息

Biomedical Sciences, Graduate School, Khon Kaen University, Khon Kaen, Thailand.

Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand.

出版信息

J Ethnopharmacol. 2017 Apr 6;201:9-16. doi: 10.1016/j.jep.2017.02.044. Epub 2017 Mar 1.

DOI:10.1016/j.jep.2017.02.044
PMID:28257978
Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Corn silk or the stigma of Zea mays L. has traditionally been used in weight loss stimulation and treatment of cystitis, urinary infections and obesity. Purple corn silk, rich of polyphenolic substances, was reported on anti-diabetic and anti-obesity effect in animal studies. However, scientific evidence on mechanisms and targets of action of purple corn silk related to adipocyte life cycle has been limited.

AIM OF STUDY

To determine phytochemical compositions and investigate anti-obesity potential of the purple corn silk focusing on interruption of adipocyte life cycle; effect on pre-adipocyte proliferation, adipogenesis, adipocyte lipolysis, and apoptosis.

MATERIALS AND METHODS

The ethanolic purple corn silk extract (PCS) was prepared and investigated for phytochemical compositions by LC/MS/MS technique and anti-obesity potential using murine 3T3-L1 cell line. Using methyl thiazole tetrazolium (MTT) assay, the effects on pre-adipocytes and adipocyte viability and on pre-adipocytes proliferation at 24-, 48-, and 72-h incubation period were evaluated. In addition, anti-adipogenesis via inhibition on adipocyte differentiation and reduction of total lipid accumulation was evaluated using Oil Red O staining and spectrophotometric methods, respectively. The lipolysis effect was determined by measurement of glycerol released content using glycerol test kit after 48-h treatment of PCS to adipocytes. Apoptosis inductive effect was done by using 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI) staining method.

RESULTS

The polyphenols including anthocyanins, quercetin and phenolic acids and derivatives were found as the major chemical compositions of the PCS. With multiple-stages interruption on the adipocyte life cycle, anti-obesity effect of PCS was interestingly demonstrated. When compared to the control, the PCS at concentration range between 250-1000 μg/mL showed anti-adipogenesis effect as expressing of significant inhibition on pre-adipocyte proliferation at all incubation period (43.52±5.28 - 75.51±9.09%) and significant decreasing of total lipid accumulation at concentration of 500μg/mL (80.22±6.58%) and 1000μg/mL (69.62±5.42%). Moreover, the PCS exhibited lipolysis and apoptosis inductive effect with dose dependent manner and significance at concentration of 1000μg/mL by increase of released glycerol content (173.88±6.13% of the control) and of nuclei condensing and apoptotic bodies (with relative apoptosis induction as 131.74±1.64% of the control).

CONCLUSION

Our data has evidenced the anti-obesity potential of PCS related interruption at multiple stages of adipocyte life cycle. Its potency was attributed to inhibition on adipocyte proliferation and adipogenesis as well as induction on lipolysis and apoptosis at high concentration. However, further in vivo investigation should be considered to insist the possibility in applications of PCS in prevention and treatment of obesity.

摘要

民族药理学相关性

玉米须,即玉米(Zea mays L.)的柱头,传统上一直用于促进减肥以及治疗膀胱炎、泌尿系统感染和肥胖症。紫色玉米须富含多酚类物质,动物研究报道其具有抗糖尿病和抗肥胖作用。然而,关于紫色玉米须与脂肪细胞生命周期相关的作用机制和靶点的科学证据有限。

研究目的

确定紫色玉米须的植物化学成分,并研究其针对脂肪细胞生命周期中断的抗肥胖潜力;研究其对前脂肪细胞增殖、脂肪生成、脂肪细胞脂解和凋亡的影响。

材料与方法

制备紫色玉米须乙醇提取物(PCS),采用液相色谱/串联质谱(LC/MS/MS)技术研究其植物化学成分,并使用小鼠3T3-L1细胞系研究其抗肥胖潜力。采用甲基噻唑四氮唑(MTT)法,评估在24、48和72小时孵育期对前脂肪细胞和脂肪细胞活力以及前脂肪细胞增殖的影响。此外,分别采用油红O染色和分光光度法评估通过抑制脂肪细胞分化和减少总脂质积累的抗脂肪生成作用。通过使用甘油测试试剂盒在PCS处理脂肪细胞48小时后测量释放的甘油含量来确定脂解作用。采用2-(4-脒基苯基)-6-吲哚甲脒二盐酸盐(DAPI)染色法进行凋亡诱导作用研究。

结果

发现多酚类物质,包括花青素、槲皮素、酚酸及其衍生物是PCS的主要化学成分。有趣的是,PCS在脂肪细胞生命周期的多个阶段均有阻断作用,从而显示出抗肥胖作用。与对照组相比,浓度在250 - 1000μg/mL之间的PCS表现出抗脂肪生成作用,即在所有孵育期对前脂肪细胞增殖均有显著抑制(43.52±5.28 - 75.51±9.09%),并且在500μg/mL(80.22±6.58%)和1000μg/mL(69.62±5.42%)浓度下总脂质积累显著减少。此外,PCS以剂量依赖性方式表现出脂解和凋亡诱导作用,在1000μg/mL浓度下具有显著性,表现为释放的甘油含量增加(为对照组的173.88±6.13%)以及细胞核浓缩和凋亡小体增加(相对凋亡诱导率为对照组的131.74±1.64%)。

结论

我们的数据证明了PCS在脂肪细胞生命周期多个阶段相关的抗肥胖潜力。其功效归因于对脂肪细胞增殖和脂肪生成的抑制以及在高浓度下对脂解和凋亡的诱导。然而,应考虑进一步进行体内研究以确定PCS在预防和治疗肥胖症方面应用的可能性。

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