Pereira W K V, Lonardoni M V C, Grespan R, Caparroz-Assef S M, Cuman R K N, Bersani-Amado C A
Laboratory of Inflammation, Department of Pharmacy and Pharmacology, University of Maringá, Avenida Colombo, 5790, 87020-900 Maringá-PR, Brazil.
J Infect. 2005 Aug;51(2):157-64. doi: 10.1016/j.jinf.2004.09.009. Epub 2004 Nov 5.
This study investigates the action of Canova medication (CM) on experimental infection by Leishmania (Leishmania) amazonensis, utilizing in vitro and in vivo assays. For the in vitro tests, Balb/c mouse peritoneal macrophages (5x10(5) cells in 500 microl of culture medium, supplemented with 10% fetal calf serum, penicillin (100 U/ml) and streptomycin (0.1 mg/ml) (were distributed in 24-well plates and CM was added at concentrations of 20 or 40%. Twenty-four hours later, the macrophages were infected with Leishmania amastigotes in culture medium. The effect of CM on macrophages leishmanicidal activity in 24 and 48 h cultures was evaluated by determining infection index and measuring nitric oxide (NO) production. The in vivo tests were performed in mice infected with 10(7)L. (L.) amazonensis promastigotes injected in to the right hind footpad (25 microl in phosphate buffered saline). The progression of the lesions was examined over a 9-week period by measuring footpad swelling, and the parasite load in regional lymph nodes and spleen. The in vitro results showed that at 40% CM reduced the infection index, and induced NO production in the elicited macrophages, which suggests that the inhibitory effect on infection index may be mediated by NO. In the in vivo infection, when administered, orally or subcutaneously in mice, CM reduced infection by L. (L.) amazonensis in the paws, resulting in smaller lesions. CM treatment also decreased parasite load in the regional popliteal lymph nodes and in the spleen. These results suggest that CM modulates experimental infection by L. (L.) amazonensis, controlling infection progression and limiting dissemination.
本研究利用体外和体内试验，研究了卡诺瓦药物（CM）对亚马逊利什曼原虫（Leishmania）实验性感染的作用。体外试验中，将Balb/c小鼠腹腔巨噬细胞（5×10⁵个细胞，置于500微升补充有10%胎牛血清、青霉素（100 U/ml）和链霉素（0.1 mg/ml）的培养基中）接种于24孔板，加入浓度为20%或40%的CM。24小时后，用培养基中的利什曼原虫无鞭毛体感染巨噬细胞。通过测定感染指数和测量一氧化氮（NO）产生，评估CM在24小时和48小时培养中对巨噬细胞杀利什曼原虫活性的影响。体内试验在感染10⁷个亚马逊利什曼原虫前鞭毛体的小鼠中进行，将其注入右后足垫（25微升于磷酸盐缓冲盐水中）。在9周时间内，通过测量足垫肿胀、区域淋巴结和脾脏中的寄生虫载量来检查病变进展。体外结果显示，40%的CM降低了感染指数，并诱导诱导型巨噬细胞产生NO，这表明对感染指数的抑制作用可能由NO介导。在体内感染中，当在小鼠中口服或皮下给药时，CM减少了亚马逊利什曼原虫在爪部的感染，导致病变较小。CM治疗还降低了区域腘淋巴结和脾脏中的寄生虫载量。这些结果表明，CM可调节亚马逊利什曼原虫的实验性感染，控制感染进展并限制传播。
