University of Lomé, Togo; University of Agricultural Science and Veterinary Medicine, Manastur Street. 3-5, 400372, Cluj-Napoca, Romania; Sree Siddaganga College of Pharmacy, B.H. Road, Tumkur, 572 102, Karnataka, India.
University of Agricultural Science and Veterinary Medicine, Manastur Street. 3-5, 400372, Cluj-Napoca, Romania.
J Ethnopharmacol. 2020 Oct 28;261:113133. doi: 10.1016/j.jep.2020.113133. Epub 2020 Jul 14.
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.
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.
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.
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.
Taken together, the present study provided the scientific footage for the traditional use of Combretum micranthum.
Combretum micranthum G. Don（CM）在整个西非的传统医学中被广泛使用，通常被称为“长寿草药茶”或“治愈植物”。此外，传统治疗师经常使用该植物来减轻肾脏疾病。
探讨标准化的 Combretum micranthum 水醇提取物对烟酰胺-链脲佐菌素诱导的糖尿病肾病大鼠的肾脏保护作用。此外，还对该植物的生物活性化合物进行了基于计算机的计算实验，以对抗 PPARα 和 PPARγ。
雄性大鼠通过单次腹腔注射 STZ（50mg/kg）诱导糖尿病，15 分钟后腹腔注射 NA（100mg/kg）溶解在生理盐水中。糖尿病大鼠每天口服 CM 提取物（200 和 400mg/kg），持续 8 周。每周测量大鼠的体重和血糖（非空腹和空腹）。还测量了大鼠的每日食物和水的摄入量。经过 8 周的治疗，测量了尿液生化参数，如 N-乙酰-β-D-氨基葡萄糖苷酶（NAG）、尿素（UR）、尿酸（UA）、肌酐（CRE）和血清糖尿病标志物、肾脏损伤和肝脏损伤的标志物，如胰岛素、脂质参数）、丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）和γ-谷氨酰转移酶（γGT）、白蛋白（Alb）、镁（Mg）、钙（Ca）、磷（P）。还估计了血液糖化血红蛋白（HbA1）。肾脏和肝脏用于生化估计氧化应激标志物，如脂质过氧化、超氧化物歧化酶（SOD）活性和谷胱甘肽过氧化物酶（GPx）活性。肾脏和胰腺用于组织病理学研究。此外，还进行了 CM 提取物的 HPLC 化学分析和基于计算机的分子模拟实验。
八周结束时，通过改变血清和尿液肾功能和肝功能标志物、氧化应激标志物和肾脏组织病理学变化，证实了长期糖尿病引起的肾脏损伤。CM 提取物的治疗改善了糖尿病诱导的肾脏生化参数和组织病理学变化。此外，HPLC-UV 和 MS 实验表明，CM 提取物含有几种生物活性化合物，包括 hyperozide（62.35μg/mg 提取物）和 quercitrin（19.07μg/mg 提取物）。基于计算机的实验显示，cianidanol（-17.133）和儿茶素（-15.107）对 PPARα 的结合评分更高，而 luteoforol（-11.038）和表没食子儿茶素（-10.736）对 PPARγ 的结合评分更高。基于对接和药物相似性评分，选择了四种生物活性化合物进行分子动力学实验。在 30 种化合物中，cianidanol 和表没食子儿茶素被认为是通过激活 PPARα 和 PPARγ 靶蛋白治疗 DN 的潜在候选药物。
综上所述，本研究为 Combretum micranthum 的传统用途提供了科学依据。
