The Experimental Research Of Effects Of Yunnan Baiyao On The Arachidonic Acid Metabolized Of 5-LOX Pathway

Objective:The aim of this study is to observe the effects of Yunnan Baiyao (YNB) on the key enzyme and inflammation cytokines of arachidonic acid metabolized of 5-LOX pathway, The results may provide laboratory evidence of the YNB used for inflammation disease.Methods:104 healthy male SD rats,7-8 week-old, were randomly divided into four groups, 26 rats each group,A:normal group,B:model group,C:mizolastine group,D:Yunnan Baiyao(YNB) group.A week before modeling, intragastric administration to four group,one time a day.Continuous intragastric administration for 7 days.A and B group intragastrically adiminstered with normal saline 1 mL,C group intragastrically adiminstered with 0.3mg/kg mizolastine 1 mL and D group intragastrically adiminstered with 50mg/ml YNB 1 mL. Two hours after intragastric administration at the seventh day, B、C and D groups were subcutaneously injected 0.1 mL arachidonic acid in the left paw,and injected 0.1 mL carbonate buffer solution in the right paw as control,the normal group received no injection.Respectively measuring paw volume after injected in 1 hour,2 hour,3 hour and 4 hour each time point.At the same time,13 rats were randomly taken from each group in 2h and 4h, put to death after collecting the blood from the abdominal aorta for serological detection.Inflammation tissues of the left and right foot paw were preserved in 4% poly formaldehyde fixative,RNA preservation solution and clean 1.5ml EP tube for histological and molecular biological. (1)histologic:HE staining was used to observe the level of inflammation cell infiltration in the inflammation tissues.(2) molecular biologic:RT-PCR was used to observe the level of 5-LOX,LTB4r,cPLA2 and FLAP mRNA relatively expression in the inflammation tissues.(3) serologic:ELISA was used to detect the level of 5-LOX,LTB4 and cPLA2 in plasma.Statistical analysis of the experimental results using SPSS 17.0 statistical software.Results:(1) The level of rat paw swelling in each time point:1 h after modeling, compared with nomal group,model group, mizolastine group and YNB group foot paw swelling was significantly higher, reached the peak at 2h. Subsequently mizolastine and YNB group swelling relieved gradually. The level of rat paw swelling relieved still higher than nomal group(P<0.05).(2) The results of ELISA showed that the level of 5-LOX,LTB4 and cPLA2 in plasma of mizolastine group and YNB group significantly lower than that of model group (P<0.05),but still higher than nomal group(P<0.05). Compared with YNB group, the factor concentration of mizolastine group was lower than that of YNB group (P< 0.05).However,there were no statistical significance in each group at the different time points(P> 0.05).(3) The RT-PCR results showed that the mRNA of 5-LOX,LTB4r,cPLA2 and FLAP in the inflammation tissue of mizolastine group and YNB group evidently lower than that of model group (P<0.05),but still higher than nomal group(P<0.05). When compared with YNB group, mizolastine group showed low-level expression(P< 0.05).However,there were no statistical significance in each group at the different time points(P> 0.05).Conclusion:mizolastine and YNB can obviously inhibited the expression of key enzyme of arachidonic acid metabolized of 5-LOX pathway.Reducing the production of inflammation cytokines and consequently relieve the swell of inflammation tissue.