Effect Of Reactive Oxygen Species On The Function Of Mast Cells In Connective Tissue

Many previous researches found that connective tissue might be the physical basis of acupuncture meridian system,and mast cell had close relationship with meridian transmission phenomenon.The distribution of reactive oxygen species(ROS)was found to have high coincidence with acupuncture meridian by fluorescent staining in our lab.So in this paper,the effect of ROS on the function of mast cell of connective tissue was analyzed,aiming to further investigate the relationship between ROS,mast cell and acupuncture meridian system.The experiment was divided into two parts:tissue staining experiment and in vitro cell culture.(1)The method of tissue staining experiment:Selecting linea alba,small intestine and mesentery of rat to carry out morphological researches;Using the staining of HE,toluidine blue to study the distribution of mast cells in connective tissues.(2)The method of in vitro cell culture:Using the method of cell culture,the effect of ROS on the function of mast cell,such as proliferation,morphology,and degranulation was analyzed;By co-culture and three-dimensional culture of mast cell and fibroblast,the interaction of the two kinds of cell and extracellular matrix was investigated.The following results were found:(1)The results of tissue staining:The result of toluidine blue staining showed that there were many mast cells in the linea alba,small intestine and mesentery,and mast cells existed around the blood vessels exhibited linear arrangement.Injecting an overdose of H2O2 into the rat caudal vein led to the degranulation of mast cells greatly,and injecting an overdose of glutathione led to the cell death.(2)The results of cell culture:Low concentration of H2O2 promoted the proliferation of mast cells;high concentration of H2O2 inhibited the proliferation,and the higher concentration of H2O2,the more obvious inhibitory effect.Paraquat in all tested concentration inhibited the proliferation of mast cells.H2O2(50?M-800 ?M)showed no significant effect on the degranulation of mast cells.While mast cells treated with high concentration of paraquat released of higher histamine than control and the treatment of SOD decreased the release.Through the co-culture of mast cell and fibroblast,it was found that with the treatment of H2O2,fibroblast could promote mast cell degranulation.While with the treatment of paraquat,fibroblast could inhibit the degranulation.By three-dimensional culture of mast cell and fibroblast,mast cells was found to promote fibroblast proliferation,spreading and lamellipodia formation,as well as to reduce the damage of ROS to fibroblasts.Conclusion:it was confirmed that there were many mast cells in the high concentration of ROS-containing connective tissue of linea alba,small intestine and mesentery;the suitable concentration of ROS was crucial for mast cells to maintain degranulation function.ROS exhibited significant effect on the mast cell proliferation,degranulation and the function of the network between mast cells and fibroblasts,thus play a possible role as a signal transducer in the connective tissue of meridian system.These results will help us to further explore the mechanism of meridian system.