ニコチンアミド-ストレプトゾトシンによる糖尿病性腎症に対するCombretum micranthum G. Donの腎保護効果.In-vivoおよびIn-silico実験

Nephroprotective effect of Combretum micranthum G. Don in nicotinamide-streptozotocin induced diabetic nephropathy in rats: In-vivo and In-silico experiments.

• Mabozou Kpemissi
• Adrian-Valentin PotÂrniche
• Povi Lawson-Evi
• Kossi Metowogo
• Mamatchi Melila
• Pare Dramane
• Marian Taulescu
• Vivek Chandramohan
• Doddamavattur Shivalingaiah Suhas
• Tumbadi Adinarayanashetty Puneeth
• Vijaya Kumar S
• Laurian Vlase
• Sanda Andrei
• Kwashie Eklu-Gadegbeku
• Bogdan Sevastre
• Veeresh P Veerapur

抄録

文化人類学的分布:

Combretum micranthum G. Don (CM)は西アフリカの伝統医学で広く使われており、一般的には「長寿のハーブティー」や「癒しの植物」として知られています。さらに、伝統的な治療者は、腎障害の緩和にこの植物を頻繁に使用しています。

ETHNOPHARMACOLOGICAL RELEVANCE: Combretum micranthum G. Don (CM) is extensively used in traditional medicine throughout West Africa and commonly known as "long-life herbal tea" or "plant to heal". Further, traditional healers frequently use the title plant to mitigate of renal disorders.

研究の目的:

ラットにおけるニコチンアミド-ストレプトゾトシン誘導糖尿病性腎症に対するCombretum micranthumの標準化されたヒドロアルコール抽出物の腎保護作用を検討した。また、表題植物の生理活性化合物を用いて、PPARαおよびPPARγに対するインシリコ計算実験を行った。

AIM OF THE STUDY: To explore the nephroprotective property of standardised hydroalcoholic extract of Combretum micranthum in nicotinamide-streptozotocin induced diabetic nephropathy in rats. In addition, in-silico computational experiments were performed with bioactive compounds of the title plant against PPARα and PPARγ.

材料と方法:

雄性ラットに、通常の生理食塩水に溶解したNA（100 mg/Kg）をIP投与した15分後に、STZ（50 mg/Kg）を単回腹腔内（ip）注射して糖尿病とした。糖尿病ラットには、ＣＭ抽出物（２００及び４００ｍｇ／ｋｇｐ.ｏ.）を毎日、８週間投与した。ラットの体重および血糖値（非絶食および空腹時）を毎週測定した。また、毎日の餌と水の消費量も測定した。処置の8週間後、尿生化学的パラメータ、例えばN-アセチル-β-D-グルコサミニダーゼ（NAG）、尿素（UR）、尿酸（UA）、クレアチニン（CRE）、インスリンなどの糖尿病、腎障害、肝障害の血清マーカー、脂質パラメータ）、アラニンアミノトランスフェラーゼ（ALT）、アスパラギン酸アミノトランスフェラーゼ（AST）、γ-グルタミルトランスフェラーゼ（γGT）、アルブミン（Alb）、マグネシウム（Mg）、カルシウム（Ca）、リン（P）などの尿生化学的パラメータを推定した。腎臓と肝臓は、脂質過酸化、スーパーオキシドディスムターゼ(SOD)活性、グルタチオンペルオキシダーゼ(GPx)活性などの酸化ストレスマーカーの生化学的推定に使用した。病理組織学的研究には腎臓と膵臓を用いた。さらに、CM 抽出物の HPLC ケモプロファイリングとインシリコ分子シミュレーション実験を行った。

MATERIAL AND METHODS: Male rats were made diabetic by a single intraperitoneal (ip) injection of STZ (50 mg/Kg), 15 min after ip administration of NA (100 mg/Kg) dissolved in normal saline. The diabetic rats received CM extract (200 and 400 mg/kg p.o.) daily, for eight weeks. Body weights and blood glucose (non-fasting and fasting) of rats were measured weekly. Daily food and water consumption were also measured. After 8 weeks of treatment, urine biochemical parameters such as N-Acetyl-β-D-Glucosaminidase (NAG), urea (UR), uric acid (UA), creatinine (CRE), and serum markers of diabetes, kidney damage and liver damage such as insulin, lipid parameters), alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma-glutamyl transferase (γGT), albumin (Alb), magnesium (Mg), calcium (Ca), phosphorus (P), were estimated. Blood glycosylated hemoglobin (HbA1) were also estimated. kidney and liver were used for biochemical estimation of oxidative stress markers such as lipid peroxidation, superoxide dismutase (SOD) activity and glutathione peroxidase (GPx) activity. The kidney and pancreas were used for histopathological study. Further, HPLC chemoprofiling of CM extract and in-silico molecular simulation experiments were performed.

結果:

RESULTS: At the end of eight weeks, renal damage induced by the consequence of prolong diabetic condition was confirmed by altered levels of serum and urine kidney and liver function markers, oxidative stress markers and histopathological variations in kidney. Treatment with CM extract ameliorated the diabetes mellitus-induced renal biochemical parameters and histopathological changes. Further, HPLC-UV & MS experiments revealed that CM extract contains several bioactive compounds including hyperozide (62.35 μg/mg of extract) and quercitrin (19.07 μg/mg of extract). In-silico experiment exhibited Cianidanol (-17.133), epicatechin (-15.107) exhibited higher docking score against PPARα and luteoforol (-11.038), epigallocatechin (-10.736) against PPARγ. Based on docking and drug likeness score, four bioactive compounds were selected for molecular dynamic experiments. Cianidanol and epigallocatechin out of the 30 compounds are concluded as a potential candidate for the treatment of DN through activating PPARα and PPARγ target protein.

結論:

本研究は、Combretum micranthumの伝統的な使用のための科学的な映像を提供した。

CONCLUSIONS: Taken together, the present study provided the scientific footage for the traditional use of Combretum micranthum.

Copyright © 2020 Elsevier B.V. All rights reserved.