Olajide Olayemi J, Enaibe Bernard U, Bankole Oluwamolakun O, Akinola Oluwole B, Laoye Babafemi J, Ogundele Olalekan M
Department of Anatomy, College of Health Sciences, University of Ilorin, Ilorin, Kwara State, Nigeria.
Neurobiology Unit; Department of Biological Sciences, College of Sciences, Afe Babalola University, Ado-Ekiti, Ekiti State, Nigeria.
Metab Brain Dis. 2016 Feb;31(1):25-35. doi: 10.1007/s11011-015-9674-0. Epub 2015 Apr 29.
Kolaviron is a phytochemical isolated from Garcina kola (G. kola); a common oral masticatory agent in Nigeria (West Africa). It is a bioflavonoid used--as an antiviral, anti-inflammatory and antioxidant--in relieving the symptoms of several diseases and infections. In this study we have evaluated the neuroprotective and regenerative effect of kolaviron in neurons of the prefrontal cortex (Pfc) before or after exposure to sodium azide (NaN3) induced oxidative stress. Separate groups of animals were treated as follows; kolaviron (200 mg/Kg) for 21 days; kolaviron (200 mg/Kg for 21 days) followed by NaN3 treatment (20 mg/Kg for 5 days); NaN3 treatment (20 mg/Kg for 5 days) followed by kolaviron (200 mg/Kg for 21 days); 1 ml of corn-oil (21 days-vehicle); NaN3 treatment (20 mg/Kg for 5 days). Exploratory activity associated with Pfc function was assessed in the open field test (OFT) following which the microscopic anatomy of the prefrontal cortex was examined in histology (Haematoxylin and Eosin) and antigen retrieval Immunohistochemistry to show astroglia activation (GFAP), neuronal metabolism (NSE), cytoskeleton (NF) and cell cycle dysregulation (p53). Subsequently, we quantified the level of Glucose-6-phosphate dehydrogenase (G6PDH) and lactate dehydrogenase (LDH) in the brain tissue homogenate as a measure of stress-related glucose metabolism. Kolaviron (Kv) and Kolaviron/NaN3 treatment caused no prominent change in astroglia density and size while NaN3 and NaN3/Kv induced astroglia activation and scar formation (astrogliosis) in the Pfc when compared with the control. Similarly, Kolaviron and Kv/NaN3 did not alter NSE expression (glucose metabolism) while NaN3 and NaN3/Kv treatment increased cortical NSE expression; thus indicating stress related metabolism. Further studies on enzymes of glucose metabolism (G6PDH and LDH) showed that NaN3 increased LDH while kolaviron reduced LDH in the brain tissue homogenate (P < 0.001). In addition kolaviron treatment before (P < 0.001) or after (P < 0.05) NaN3 treatment also reduced LDH expression; thus supporting its role in suppression of oxidative stress. Interestingly, NF deposition increased in the Pfc after kolaviron treatment while Kv/NaN3 showed no significant change in NF when compared with the control. In furtherance, NaN3 and NaN3/Kv caused a decrease in NF deposition (degeneration). Ultimately, the protective effect of KV administered prior to NaN3 treatment was confirmed through p53 expression; which was similar to the control. However, NaN3 and NaN3/Kv caused an increase in p53 expression in the Pfc neurons (cell cycle dysregulation). We conclude that kolaviron is not neurotoxic when used at 200 mg/Kg BW. Furthermore, 200 mg/Kg of kolaviron administered prior to NaN3 treatment (Kv/NaN3) was neuroprotective when compared with Kolaviron administered after NaN3 treatment (NaN3/Kv). Some of the observed effects of kolaviron administered before NaN3 treatment includes reduction of astroglia activation, absence of astroglia scars, antioxidation (reduced NSE and LDH), prevention of neurofilament loss and cell cycle regulation.
可乐维隆是从可乐果(Garcina kola)中分离出的一种植物化学物质;可乐果是尼日利亚(西非)一种常见的口腔咀嚼剂。它是一种生物类黄酮,用作抗病毒、抗炎和抗氧化剂,可缓解多种疾病和感染的症状。在本研究中,我们评估了可乐维隆在暴露于叠氮化钠(NaN3)诱导的氧化应激之前或之后对前额叶皮质(Pfc)神经元的神经保护和再生作用。将动物分为不同组并进行如下处理:给予可乐维隆(200毫克/千克),持续21天;先给予可乐维隆(200毫克/千克,持续21天),随后给予NaN3处理(20毫克/千克,持续5天);先给予NaN3处理(20毫克/千克,持续5天),随后给予可乐维隆(200毫克/千克,持续21天);给予1毫升玉米油(持续21天,作为对照);给予NaN3处理(20毫克/千克,持续5天)。在旷场试验(OFT)中评估与Pfc功能相关的探索性活动,之后在组织学(苏木精和伊红染色)和抗原修复免疫组织化学中检查前额叶皮质的微观解剖结构,以显示星形胶质细胞活化(GFAP)、神经元代谢(NSE)、细胞骨架(NF)和细胞周期失调(p53)。随后,我们定量测定脑组织匀浆中葡萄糖-6-磷酸脱氢酶(G6PDH)和乳酸脱氢酶(LDH)的水平,作为应激相关葡萄糖代谢的指标。与对照组相比,可乐维隆(Kv)和可乐维隆/NaN3处理组的星形胶质细胞密度和大小没有显著变化,而NaN3组和NaN3/Kv组诱导了Pfc中的星形胶质细胞活化和瘢痕形成(星形胶质细胞增生)。同样,可乐维隆和Kv/NaN3组未改变NSE表达(葡萄糖代谢),而NaN3组和NaN3/Kv处理组增加了皮质NSE表达;因此表明存在应激相关代谢。对葡萄糖代谢酶(G6PDH和LDH)的进一步研究表明,NaN3增加了脑组织匀浆中的LDH,而可乐维隆降低了LDH(P < 0.001)。此外,在NaN3处理之前(P < 0.001)或之后(P < 0.05)给予可乐维隆处理也降低了LDH表达;因此支持其在抑制氧化应激中的作用。有趣的是,可乐维隆处理后Pfc中的NF沉积增加,而与对照组相比,Kv/NaN3组的NF没有显著变化。此外,NaN3组和NaN3/Kv组导致NF沉积减少(变性)。最终,通过p53表达证实了在NaN3处理之前给予KV的保护作用;这与对照组相似。然而,NaN3组和NaN3/Kv组导致Pfc神经元中p53表达增加(细胞周期失调)。我们得出结论,以200毫克/千克体重使用可乐维隆时没有神经毒性。此外,与在NaN3处理之后给予可乐维隆(NaN3/Kv)相比,在NaN3处理之前给予200毫克/千克的可乐维隆(Kv/NaN3)具有神经保护作用。在NaN3处理之前给予可乐维隆所观察到的一些作用包括减少星形胶质细胞活化、不存在星形胶质细胞瘢痕、抗氧化(降低NSE和LDH)、预防神经丝丢失和细胞周期调节。