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L. 对分离的胰腺细胞中葡萄糖摄取的作用以及对3T3-L1细胞系中脂肪细胞分化的抑制作用。

Role of L. on Glucose Uptake in Isolated Pancreatic Cells and Inhibition of Adipocyte Differentiation in 3T3-L1 Cell Line.

作者信息

Paul Tania, Apte Kishori G, Parab Pradeep B, Das Biswadeep

机构信息

APT Research Foundation, Pune, Maharashtra, India.

APT Testing and Research Pvt. Ltd., Pune, Maharashtra, India.

出版信息

Pharmacogn Mag. 2017 Jul;13(Suppl 2):S334-S338. doi: 10.4103/pm.pm_415_16. Epub 2017 Jul 11.

Abstract

BACKGROUND

(AP) is a pteridophyte that shows antihyperglycemic activity diabetic model, but the mechanism of action is unknown.

OBJECTIVE

AP was found to play a pivotal role in minimizing the high blood glucose in alloxan-induced diabetic rats. Simultaneously, it was observed that it could maintain the normal lipid profile even in diabetic condition. To investigate its insulin-like activity along with its inhibitory role on adipocyte differentiation became the objective of our present study.

MATERIALS AND METHODS

Glucose uptake potential of this fern was done in isolated pancreatic islets and inhibition of adipocyte differentiation was assessed in 3T3-L1 cell line. Before this, the cytotoxic concentration was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay on L929 cell line. To determine its role in lipid metabolism, the oil droplets produced in adipocytes were stained with Oil 'O' red staining, and triglyceride levels of various drug treatments were measured spectrophotometrically.

RESULTS

This fern extract was found to be actively utilizing glucose in the glucose uptake assay. Moreover, it was also involved in inhibiting differentiation of pro-adipocyte to adipocyte in the 3T3-L1 cell lines. The percentage inhibition as obtained from the absorbance showed that the ethanolic extract at the concentration of 200 μg/ml showed 32.48% inhibition.

CONCLUSION

All the above-mentioned parameters when appraised indicated that this fern could be used as an alternative medicine in managing diabetes associated with obesity.

SUMMARY

Adiantum phillippense (AP) is a pteridophyte that can work as antihyperglycemic agent by minimizing some adverse effects produced by diabetes. Diabetes produces oxidative stress, hampers normal glucose uptake in the pancreas, promotes adipocyte differentiation, and leads to obesity, and as a result, it generates catastrophic effect to the normal cells. The present study has shown that ethanolic extract of AP gives better protection rate against H O-induced cytotoxicity, elicits insulinotropic activity in isolated mouse pancreatic glucose uptake assay. It also inhibits the preadipocytes to become mature adipocytes judged by morphology or lipid-specific Oil-Red-O staining of 3T3-L1 cell line. AP: Adiantum phillipense; MTT: (3-(4,5- Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide); BSA: Bovine serum albumin; FCS: Fetal calf serum; DMEM: Dulbecco's minimum essential media; RPMI: Roswell park memorial institute medium; DTZ: Dithizone; TG: Triglyceride; PPARγ: Peroxisome proliferator-activated receptor gamma; IBMX: 3-isobutyl-1-methylxanthine; nm: Nanometer; GI: Growth Inhibition; ELISA: Enzyme linked immunosorbent assay.

摘要

背景

扇叶铁线蕨是一种显示出抗高血糖活性的蕨类植物,可用于糖尿病模型,但作用机制尚不清楚。

目的

发现扇叶铁线蕨在减轻四氧嘧啶诱导的糖尿病大鼠高血糖方面起关键作用。同时,观察到即使在糖尿病状态下它也能维持正常的血脂水平。研究其胰岛素样活性及其对脂肪细胞分化的抑制作用成为本研究的目的。

材料与方法

在分离的胰岛中检测这种蕨类植物的葡萄糖摄取潜力,并在3T3-L1细胞系中评估其对脂肪细胞分化的抑制作用。在此之前,通过对L929细胞系进行3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐测定来确定细胞毒性浓度。为了确定其在脂质代谢中的作用,用油红O染色对脂肪细胞中产生的油滴进行染色,并通过分光光度法测量各种药物处理的甘油三酯水平。

结果

在葡萄糖摄取试验中发现这种蕨类植物提取物能积极利用葡萄糖。此外,它还参与抑制3T3-L1细胞系中前脂肪细胞向脂肪细胞的分化。从吸光度获得的抑制百分比表明,浓度为200μg/ml的乙醇提取物显示出32.48%的抑制率。

结论

上述所有参数评估表明,这种蕨类植物可作为治疗与肥胖相关糖尿病的替代药物。

总结

扇叶铁线蕨是一种蕨类植物,可通过减轻糖尿病产生的一些不良影响来作为抗高血糖药物。糖尿病会产生氧化应激,阻碍胰腺中正常的葡萄糖摄取,促进脂肪细胞分化,并导致肥胖,因此,它对正常细胞产生灾难性影响。本研究表明,扇叶铁线蕨的乙醇提取物对过氧化氢诱导的细胞毒性具有更好的保护率,在分离的小鼠胰腺葡萄糖摄取试验中具有促胰岛素活性。通过3T3-L1细胞系的形态学或脂质特异性油红O染色判断,它还能抑制前脂肪细胞成为成熟脂肪细胞。AP:扇叶铁线蕨;MTT:3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐;BSA:牛血清白蛋白;FCS:胎牛血清;DMEM:杜氏改良Eagle培养基;RPMI:罗斯威尔公园纪念研究所培养基;DTZ:双硫腙;TG:甘油三酯;PPARγ:过氧化物酶体增殖物激活受体γ;IBMX:3-异丁基-1-甲基黄嘌呤;nm:纳米;GI:生长抑制;ELISA:酶联免疫吸附测定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2fd0/5538176/3710bc69036f/PM-13-334-g001.jpg

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