Effect of neem leaf extract (Azadirachta indica) in reducing the degree of parasitemia and apoptosis in C57BL mice with cerebral malaria

Z. Rahmah, Kautsar Citra Nirmala, Achmad Nashichuddin, Riskiyana Riskiyana, Alvi Milliana, Nurfianti Indriana, L. Astari, Prida Ayudianti, Munawar Kholil
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Abstract

Background and Aim: Brain malaria, which results from Plasmodium falciparum infection, is responsible for substantial fatalities and health issues. These processes, including cytoadherence, rosetting, and sequestration, induce an immune response, hypoxia, brain microvascular obstruction, disruption of the blood-brain barrier, and cell death. Parasitemia level can reveal the presence of infection and its association with apoptosis-related genes. Neem (Azadirachta indica) leaves with antimalarial properties could replace ineffective Indonesian malaria medications. This study was designed to evaluate the impact of neem leaf extract on cerebral malaria-induced parasitemia and neuron cell apoptosis in mice through an in vivo approach. Materials and Methods: 13–16 weeks old C57BL mice received infection by Plasmodium berghei strain ANKA. Parasitemia was estimated daily from the mice’s tail blood. 8 mg, 12 mg, and 16 mg of a 96% ethanolic neem leaf extract were orally given for 6 days. Healthy, positive, and negative controls were included for treatment comparisons. On the 7th day, brain tissue was analyzed for (p > 0.05) gene expression. Through immunohistochemistry, both cell apoptosis in neurons expressing caspase-3 within a brain sample and the degree of parasitemia in a blood smear were assessed. The Pearson correlation test and one-way analysis of variance were employed to analyze the data. Results: Neem leaf extract reduces parasitemia and neuron cell apoptosis at multiple dosages (p < 0.000). Apoptosis in brain neurons and parasitemia show a strong positive correlation (r = +0.939). Neem leaf extract at doses of 12 and 16 mg was the most effective in reducing parasitemia levels and causing cell death. Conclusion: Neem leaf therapy significantly reduced the degree of parasitemia and cell apoptosis in C57BL mice compared with the control group without treatment (p = 0.05). This shows that neem leaves have the potential to be a candidate drug for malaria. Keywords: apoptosis, Azadirachta indica, cerebral malaria, neem leaves, parasitemia.
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印楝叶提取物(Azadirachta indica)对降低 C57BL 脑疟疾小鼠寄生虫血症和细胞凋亡程度的影响
背景和目的:恶性疟原虫感染导致的脑疟疾造成大量死亡和健康问题。这些过程包括细胞粘附、沉降和螯合,诱发免疫反应、缺氧、脑微血管阻塞、血脑屏障破坏和细胞死亡。寄生虫血症水平可揭示是否存在感染及其与细胞凋亡相关基因的联系。印楝(Azadirachta indica)叶片具有抗疟特性,可以取代印尼无效的疟疾药物。本研究旨在通过体内方法评估印楝叶提取物对小鼠脑疟疾诱导的寄生虫血症和神经元细胞凋亡的影响。材料与方法:13-16 周大的 C57BL 小鼠感染了疟原虫 ANKA 株。每天从小鼠尾部血液中估计寄生虫血症。连续 6 天口服 8 毫克、12 毫克和 16 毫克 96% 的乙醇楝树叶提取物。健康对照组、阳性对照组和阴性对照组被纳入治疗比较中。第 7 天,对脑组织进行基因表达分析(p > 0.05)。通过免疫组化,评估脑样本中表达 caspase-3 的神经元的细胞凋亡情况和血液涂片中的寄生虫血症程度。数据分析采用了皮尔逊相关检验和单因素方差分析。结果印楝叶提取物在多种剂量下都能减少寄生虫血症和神经元细胞凋亡(p < 0.000)。脑神经元细胞凋亡与寄生虫血症呈强正相关(r = +0.939)。苦楝叶提取物在 12 毫克和 16 毫克剂量下对降低寄生虫血症水平和导致细胞死亡最有效。结论与未接受治疗的对照组相比,楝树叶疗法能明显降低 C57BL 小鼠的寄生虫血症和细胞凋亡程度(p = 0.05)。这表明楝树叶有可能成为治疗疟疾的候选药物。关键词:细胞凋亡;印度楝;脑疟疾;印楝叶;寄生虫血症。
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