4-methyl-umbelliferone Of Silkworm Organization Redox And Immune System

4-Methylumbelliferone (4-MU), a derivative of coumarin, has anti-tumor ability atcell level and anti-bacteria capacity in vitro. But there is few reports about the effects of4-MU on redox system and immune system of animals. Our research, used the Bombyxmori as the model animal, investigated the effect of4-MU in redox and immune systemof Bombyx mori through biochemistry measurements, expression analysis of immunegene, HPLC technology, tissue culture in vitro. We discussed the mechanism of4-MUin Bombyx mori briefly. Our results provide the theoritical foundations of further usingumbelliferone to treat tumor. The main results are following:1. The chemism on improving anti-oxygen capacity of digestive tract redoxsystem of Bombyx mori after explosure of4-MU.The Bombyx mori on Day3of the5th instar would not death after explosure4mM4-MU. And the percentage of pupation had no effect by4-MU as well. We used HPLCtechnology and found that the digestive tract tissues contained4-MU original medicineafter8min of explosure of4-MU through mouth, which indicated that4-MU couldentry in digestive tract rapidly. The result of ROS stained experiment showed that4-MUincreased ROS level in digestive tract, and the main kind of ROS is hydrogen peroxide.The T-AOC (about1.5-fold)and the capability of inhibition of SAFR(about2fold) alsowere improved. In addition, the CAT and GPx activities also increased which have theability of decomposed hydrogen peroxide in the tissue(about2-2.5-fold). NADPH alsopaly an important subsidiarity role in the course of eliminating of ROS. Further more,explosure E.coli after explosed4-MU to Bombyx mori though mouth, the antibacteriacapacity of digestive tract can be improved.Through the paraffin section of HE stainingand DAPI staining observation, we found that4-MU made no damages to the cells ofthe digestive tract. We investigated the expression levels of Mek, Erk, p38and Junsignificantly improved after administrated4-MU in digestive tract, without thediscovery of apoptosis triggered in tissue.2.4-MU intrigerred the changes of redox system in fat body of Bombyx mori.Larvae (3rd day of the5th instar) were orally exposed to4mM4-MU, anumbelliferone, which swiftly induced the generation of a large number of ROS (e.g. H2O2increased6to8-fold), and4-MU was detected in the fat body8min afteradministration. In addition, the activities of CAT and GPx were up-regulated4to11-fold and2to16-fold, respectively, and were helpful in defending fat body cellsagainst oxidative injury in combination with NADPH. Furthermore, significantincreases in the contents of T-AOC (up to approximately1-fold), antioxidants ofASAFR (by1to4-fold) and GSH were detected.Through the paraffin section of HE staining and DAPI staining observation, wefound that4-MU made no damages to the cells of the fat body. Utilizing the RT-PCRmethods, we measure the expression levels of immune-related genes (such asantibacterial peptides), the CecropinA and Moricin were inducible expressed in differentdegrees. While the expression levels of Mek、Erk、Jun were decreased in fat body afterexploring4-MU. The mechanism of action of these genes may different from which indigestive tract were. The above experimental results showed that4-MU althoughinduced a large amount of ROS, but also improved the ability of decomposition of ROSin tissues, protecting organization from damaged.3.4-MU decreased the capacity of secretion of haemolymph significantlyreduced of Bombyx moriWe used HPLC technology and found that the haemolymph contained4-MUoriginal medicine after8min of explosure of4-MU through mouth. Through thehaemolymph AO-PI staining, we found dead cell proportion in the haemolymph hasimproved greatly. By using RT-PCR method to detect the expression levels of geneswhich belong in MAPK signaling pathways,we found Mek, Erk, Jun was significantinhibited, and p38had a great increased. Therefore, we speculated that the increasednumber of death cells in haemolymph may not be caused by apoptosis. To culturehematopoietic organs in vitro, we found that administration of4-MU made the capacityof secretion of haemolymph significantly reduced, but there were no atrophy of thehematopoietic organs in the process of culture in vitro. These results shows thepropotion of dead cells in haemolymph rised may be due to reducing the the amount ofblood cells. We found an increased level of reactive oxygen species in the hematopoieticorgans after4-MU exposed through ROS staining, this may be one of the reasons causethe loss of capacity of secretion of blood cells. In addition, the expression levels ofantibacterial peptide gene(such as CecropinA, Moricin) in the haemolymph increased significantly after explosure of4-MU, while the expression levels of phagocytic-relatedgenes (Tetraspanin, ActinA1, Ced6) have no obvious impacts by4-MU.