Grillo Luciano A M, Majerowicz David, Gondim Katia C
Instituto de Bioquímica Médica, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
Insect Biochem Mol Biol. 2007 Jun;37(6):579-88. doi: 10.1016/j.ibmb.2007.03.002. Epub 2007 Mar 12.
The utilization of dietary lipids was studied in adult females of Rhodnius prolixus with the use of radiolabeled triacylglycerol (TAG). It was shown that (3)H-triolein, when added to blood meal, was hydrolyzed to free fatty acids in the posterior midgut lumen. Subsequently, free fatty acids were absorbed by posterior midgut epithelium and used in the synthesis of phospholipids, diacylglycerol (DAG) and TAG. Phospholipids, DAG and free fatty acids were then found in hemolymph, from where they were rapidly cleared, and label was found in the fat body, mainly associated with TAG. Radioactive lipids, especially TAG and phospholipids, also accumulated in the ovaries. The TAG-lipase activities of posterior midgut luminal content and tissue were characterized by incubation of these samples with (3)H-triolein in the presence of the detergent Triton X-100 and determination of the amounts of released radioactive free fatty acids. Under the conditions employed here, the release of free fatty acids was proportional to the incubation time and to the amount of sample obtained from insect midgut (enzyme source) that was added. TAG-lipase activities were affected by pH and posterior midgut tissue showed optimum activity around pH 7.0-7.5, but the luminal content had the highest activities as pH decreased. Differences in activities were observed according to calcium concentration in the medium. TAG-lipase activities were also affected by the concentration of NaCl and were activated in the presence of increasing salt concentrations. These activities were inhibited by phenylmethylsulphonyl fluoride (PMSF). On the second day after blood meal, when digestion is very intense, TAG-lipase activities were maximal and then gradually decreased.
利用放射性标记的三酰甘油(TAG)对成年雌性南美锥蝽的膳食脂质利用情况进行了研究。结果表明,添加到血餐中的(3)H-三油精在后肠中段肠腔内被水解为游离脂肪酸。随后,游离脂肪酸被后肠上皮吸收,并用于合成磷脂、二酰甘油(DAG)和TAG。然后在血淋巴中发现了磷脂、DAG和游离脂肪酸,它们从血淋巴中迅速清除,并且在脂肪体中发现了标记,主要与TAG相关。放射性脂质,尤其是TAG和磷脂,也在卵巢中积累。通过在去污剂Triton X-100存在的情况下将这些样品与(3)H-三油精一起孵育,并测定释放的放射性游离脂肪酸的量,对后肠中段肠腔内容物和组织的TAG脂肪酶活性进行了表征。在此处采用的条件下,游离脂肪酸的释放与孵育时间以及从昆虫中肠(酶源)获得并添加的样品量成正比。TAG脂肪酶活性受pH影响,后肠组织在pH 7.0-7.5左右显示出最佳活性,但随着pH降低肠腔内容物具有最高活性。根据培养基中的钙浓度观察到活性差异。TAG脂肪酶活性也受NaCl浓度影响,并且在盐浓度增加时被激活。这些活性被苯甲基磺酰氟(PMSF)抑制。在血餐摄入后的第二天,当消化非常强烈时,TAG脂肪酶活性最高,然后逐渐降低。