Department of Biology, Federal Rural University of Pernambuco, Recife, PE, Brazil.
Department of Veterinary Medicine, Federal Rural University of Pernambuco, Recife, PE, Brazil.
Int J Biol Macromol. 2021 Feb 28;171:37-43. doi: 10.1016/j.ijbiomac.2020.12.172. Epub 2021 Jan 6.
Anti-inflammatory properties have been attributed to latex proteins of the medicinal plant Calotropis procera.
A mixture of cysteine peptidases (LPp2) from C. procera latex was investigated for control of inflammatory mediators and inflammation in a mouse model of Salmonella infection.
LPp2 peptidase activity was confirmed by the BANA assay. Cytotoxicity assays were conducted with immortalized macrophages. Peritoneal macrophages (pMØ) from Swiss mice were stimulated with lipopolysaccharide (LPS) in 96-well plates and then cultured with nontoxic concentrations of LPp2. Swiss mice intravenously received LPp2 (10 mg/kg) and then were challenged intraperitoneally with virulent Salmonella enterica Ser. Typhimurium.
LPp2 was not toxic at dosages lower than 62.2 μg/mL. LPp2 treatments of pMØ stimulated with LPS impaired mRNA expression of pro-inflammatory cytokines IL-1β, TNF-α, IL-6 and IL-10. LPp2 increased the intracellular bacterial killing in infected pMØ. Mice given LPp2 had a lower number of leukocytes in the peritoneal cavity in comparison to control groups 6 h after infection. The bacterial burden and histological damage were widespread in target organs of mice receiving LPp2.
We conclude that LPp2 contains peptidases with strong anti-inflammatory properties, which may render mice more susceptible to early disseminated infection caused by Salmonella.
药用植物白花牛角瓜中的抗炎蛋白具有抗炎特性。
研究白花牛角瓜乳胶中的混合半胱氨酸肽酶（LPp2）对沙门氏菌感染小鼠模型中炎症介质和炎症的控制作用。
通过 BANA 测定法确认 LPp2 肽酶活性。用永生化巨噬细胞进行细胞毒性测定。从小鼠腹膜中分离出的巨噬细胞（pMØ）在 96 孔板中用脂多糖（LPS）刺激，然后用无毒浓度的 LPp2 培养。瑞士小鼠静脉内给予 LPp2（10mg/kg），然后用强毒沙门氏菌血清型 Typhimurium 攻击腹膜。
LPp2 的剂量低于 62.2μg/mL 时没有毒性。LPp2 处理 LPS 刺激的 pMØ 可损害促炎细胞因子 IL-1β、TNF-α、IL-6 和 IL-10 的 mRNA 表达。LPp2 增加了感染的 pMØ 中的细胞内细菌杀伤。与对照组相比，感染后 6 小时给予 LPp2 的小鼠腹腔内白细胞数量更少。接受 LPp2 的小鼠的靶器官中细菌负荷和组织学损伤广泛。
我们得出结论，LPp2 含有具有强大抗炎特性的肽酶，这可能使小鼠更容易受到沙门氏菌引起的早期播散性感染。
