Background: Protease inhibitors have been isolated from plants and present several biological activities, including immunomodulatory action.

Objective: This work aimed to evaluate a Moringa oleifera flower trypsin inhibitor (MoFTI) for acute toxicity in mice, hemolytic activity on mice erythrocytes and immunomodulatory effects on mice splenocytes.

Methods: The acute toxicity was evaluated using Swiss female mice that received a single dose of the vehicle control or MoFTI (300 mg/kg, i.p.). Behavioral alterations were observed 15–240 min after administration, and survival, weight gain, and water and food consumption were analyzed daily. Organ weights and hematological parameters were analyzed after 14 days. Hemolytic activity of MoFTI was tested using Swiss female mice erythrocytes. Splenocytes obtained from BALB/c mice were cultured in the absence or presence of MoFTI for the evaluation of cell viability and proliferation. Mitochondrial membrane potential (Δψm) and reactive oxygen species (ROS) levels were also determined. Furthermore, the culture supernatants were analyzed for the presence of cytokines and nitric oxide (NO).

Results: MoFTI did not cause death or any adverse effects on the mice except for abdominal contortions at 15–30 min after administration. MoFTI did not exhibit a significant hemolytic effect. In addition, MoFTI did not induce apoptosis or necrosis in splenocytes and had no effect on cell proliferation. Increases in cytosolic and mitochondrial ROS release, as well as Δψm reduction, were observed in MoFTI-treated cells. MoFTI was observed to induce TNF-α, IFN-γ, IL-6, IL-10, and NO release.

Conclusion: These results contribute to the ongoing evaluation of the antitumor potential of MoFTI and its effects on other immunological targets.

背景：蛋白酶抑制剂已从植物中分离出来并具有多种生物活性，包括免疫调节作用。

目的：本工作旨在评估辣木花胰蛋白酶抑制剂（MoFTI）对小鼠的急性毒性、对小鼠红细胞的溶血活性和对小鼠脾细胞的免疫调节作用。

方法：使用接受单剂量载体对照或 MoFTI（300 mg/kg，ip）的瑞士雌性小鼠评估急性毒性。给药后 15-240 分钟观察到行为改变，每天分析存活率、体重增加以及水和食物消耗。14 天后分析器官重量和血液学参数。使用瑞士雌性小鼠红细胞测试 MoFTI 的溶血活性。在不存在或存在 MoFTI 的情况下培养从 BALB/c 小鼠获得的脾细胞，以评估细胞活力和增殖。还测定了线粒体膜电位 (Δψm) 和活性氧 (ROS) 水平。此外，分析培养上清液中是否存在细胞因子和一氧化氮 (NO)。

结果：除给药后 15-30 分钟出现腹部扭曲外，MoFTI 未对小鼠造成死亡或任何不良影响。MoFTI 没有表现出显着的溶血作用。此外，MoFTI 不诱导脾细胞凋亡或坏死，对细胞增殖没有影响。在 MoFTI 处理的细胞中观察到细胞溶质和线粒体 ROS 释放的增加以及 Δψm 的减少。观察到 MoFTI 诱导 TNF-α、IFN-γ、IL-6、IL-10 和 NO 释放。

结论：这些结果有助于持续评估 MoFTI 的抗肿瘤潜力及其对其他免疫靶点的影响。