In vivo Antiplasmodial Assessment of Phyllanthus odontadenius against Plasmodium berghei in NMRI Mice

R. Nakweti, Oscar Nsendo Mwangu-Kabi, Nkembolo Nkenda Cadette, S. Ndiku
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Abstract

Aims: Phyllanthus odontadenius is one of the genus Phyllanthus species, used for number diseases treatment including malaria. Malaria today poses a real public health problem for more than hundred countries, afflicted millions people and killed an estimated 405 000 in the World. The chemoresistance of Plasmodium falciparum to modern antimalarials which are either expensive, toxic or ineffective leads to the search for new antimalarials at lower cost, non-toxic and effective within plant biodiversity. Given the interest in various P. odontadenius crude extracts in vitro on P. falciparum, an in vivo study seems necessary in order to judge the extracts effectiveness of this plant. This study therefore aims to find justification for P. odontadenius secondary metabolites antiplasmodial activity which would have been revealed in the three samples from three different province sites. Study Design: Plants P. odontadenius samples from three province harvested and dried separately, plant phytochemistry screening realized, extracts preparation for antiplasmodial test, mice parasitization with P. berghei strain, orally administration drug; Parasiteamia determination using an immersion microscope. Place and Duration of Study: Department of Radiobiology, Applied Microbiology section, General Atomic Energy Commission, Regional Nuclear Studies Center of Kinshasa. MPI and pharmacognosy laboratories in the National Biomedical Research Institute (INRB). This work took place over the period from October 11, 2020 to March 12, 2021. Methodology: Phytochemical screening P. odontadenius samples was previously determined with the chemical reagent reactions and TLC. Then, P. odontadenius methanol extracts from aerial parts harvested in three sites (Kinshasa, Kasangulu and Kwango-bridge) were administered to test mice (12.5 mg/kg and 25 mg//kg bw) after infected mice with the Plasmodium berghei strain. DMSO 10% and quinine 10 mg/kg bw were also used as controls for comparison with the samples of P. odontadenius extracts. After 5 days, parasitemia of each test and controls mice was determined. Percent of parasitemia, parasite density and percent of inhibition were calculated. Finally, the effect dose 50 of each P. odontadenius specimen was finally determined. Results: Parasiteamia rates of negative control (DMSO 10%) was high (69.98±15.03%) comparing to positive control (27.43±11.46%) and tested mice with P. odontadenius extracts (12.5 and 25 mg/kg bw) which percent’s varied from 24.66±15.84% to 59.01±22.44%. Negative control presented high parasite density with 11,342 (±2,436) comparing to the positive control (4,447±1,857) and all P. odontadenius methanol extracts which varied from 3,995±2,343 for 25 mg/kg bw to 9,570±3,319 for 12.5 mg/kg bw. Parasiteamia reduction rates followed inversely parasite density, thus, Po3 25 mg/kg bw had high parasiteamia reduction rate (65.23%) comparing to positive control with 61.32% and to P. odontadenius methanol extracts. Po3 presented 2.44 mg/kg bw as effect dose 50 comparing to Po1 (2.93 mg/kg bw) and Po2 (2.68 mg/kg bw). Males mice were highly affected to P. berghei than the females. Conclusion: This study revealed that all specimens of P. odontadenius had good in vivo antiplasmodial activities on P. berghei. All P. odontadenius extracts showed good parasitaemia inhibition compared to negative control, but P. odontadenius from Kwango-brigde (Po3) presented good behavior concerning in vivo antiplasmodial activity in comparison to P. odontadenius from Kasangulu (Po2) and that from Kinshasa (Po1). However, further studies are necessary on the in vivo toxicity of the plant and on the medicinal form that could be applied.
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在 NMRI 小鼠体内评估叶黄素对疟原虫的抗疟作用
目的:Phyllanthus odontadenius 是 Phyllanthus 属物种之一,可用于治疗包括疟疾在内的多种疾病。如今,疟疾已成为一百多个国家的真正公共卫生问题,影响着数百万人,估计有 405 000 人因此丧生。恶性疟原虫对昂贵、有毒或无效的现代抗疟药物具有化学抗药性,这促使人们在植物生物多样性范围内寻找成本低、无毒且有效的新抗疟药物。鉴于各种奥登仙翁粗提取物在体外对恶性疟原虫的作用,似乎有必要进行体内研究,以判断这种植物提取物的有效性。因此,本研究旨在为奥杜鹃次生代谢物的抗疟活性寻找依据,并在三个不同省份的三个样本中进行揭示。研究设计:分别收获和干燥来自三个省的 P. odontadenius 植物样本,进行植物化学筛选,制备提取物用于抗疟试验,用伯格氏疟原虫菌株寄生小鼠,口服给药;使用浸入式显微镜测定寄生虫。研究地点和时间:金沙萨地区核研究中心原子能总委员会应用微生物部放射生物学系。国家生物医学研究所(INRB)的 MPI 和药物学实验室。工作时间:2020 年 10 月 11 日至 2021 年 3 月 12 日。方法学:通过化学试剂反应和 TLC 对 P. odontadenius 样品进行植物化学筛选。然后,从三个地点(金沙萨、卡桑古鲁和光古桥)收获的气生部分中提取 P. odontadenius 甲醇萃取物,在小鼠感染疟原虫菌株后给试验小鼠服用(12.5 毫克/千克和 25 毫克/千克体重)。10% 二甲基亚砜和 10 毫克/千克体重的奎宁也被用作对照组,以便与奥杜鹃提取物样本进行比较。5 天后,测定每只试验小鼠和对照组小鼠的寄生虫血症。计算寄生虫血症百分比、寄生虫密度和抑制百分比。最后,确定每种 P. odontadenius 标本的作用剂量 50。结果与阳性对照组(27.43±11.46%)相比,阴性对照组(DMSO 10%)的寄生虫血症率较高(69.98±15.03%),而使用 P. odontadenius 提取物(12.5 和 25 毫克/千克体重)的受试小鼠的寄生虫血症率则从 24.66±15.84% 到 59.01±22.44%不等。与阳性对照(4 447±1 857)和所有 P. odontadenius 甲醇提取物(25 毫克/千克体重)相比,阴性对照的寄生虫密度较高,为 11 342(±2 436)只,而阳性对照的寄生虫密度则从 3 995±2 343(25 毫克/千克体重)到 9 570±3 319(12.5 毫克/千克体重)不等。寄生虫数量减少率与寄生虫密度成反比,因此,Po3 25 mg/kg 体重的寄生虫数量减少率较高(65.23%),而阳性对照为 61.32%,P. odontadenius 甲醇提取物的寄生虫数量减少率也较高。与 Po1(2.93 毫克/千克体重)和 Po2(2.68 毫克/千克体重)相比,Po3 的有效剂量 50 为 2.44 毫克/千克体重。与雌性小鼠相比,雄性小鼠对柏氏杆菌的影响更大。结论这项研究表明,所有奥氏腺狰狞豹标本对伯格氏疟原虫都具有良好的体内抗疟活性。与阴性对照组相比,所有奥氏腺狰狞萃取物都对寄生虫血症有良好的抑制作用,但与卡桑古鲁(Po2)和金沙萨(Po1)的奥氏腺狰狞萃取物相比,来自 Kwango-brigde 的奥氏腺狰狞萃取物(Po3)在体内抗疟活性方面表现良好。不过,有必要对该植物的体内毒性和可应用的药用形式进行进一步研究。
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