Effect Of Huanglianwendan In Macrophage Apoptosis In ER Stress Pathway

OjectiveAtherosclerosis(AS)is one of the main diseases that danger human health.It is also the pathology substructure of cardiovascular disease.The apoptosis of macrophage is a persistent factor in the whole process of atherosclerosis.Macrophages that apoptosised form the necrotic core of the atherosclerotic plaque,which take an important part in the instability of necrosis,myocardial infarction,stroke and peripheral vascular disease.So the investigation in the mechanisms of macrophages apoptosis could be an important potential way for anti-AS.The experiences of TCM for treating cardiovascular and cerebrovascular diseases are abundant.And according to researches and reports,the phlegm-heat is an important and common cause of atherosclerosis(AS)in clinic.Huanglianwendan Medicinal Broth is good at clearing phlegm-heat, regulating the Qi and stabilizing the mind.Researches have revealed that the clinical effects of Huanglianwendan Medicinal Broth to cure coronary heart disease are significantly.In this study,macrophages arecultured in vitro and are inducted by artificial method to apoptosis in ER stress pathway,and then the effect of Huanglianwendan Medicinal Broth is observed.Method1.The culture of RAW264.7.2.The apoptosis of RAW264.7 conducted by Tg and Fuc. 3.Preparation of the serum contains Huanglianwendan and the control serum.4.The effect of serum contains Huanglianwendan on the proliferation of RAW264.7.5.Huanglianwendan serum's effect on the macrophage apoptosis.6.Huanglianwendan serum's effect on the expression of scavenger receptor in macrophage.7.Statistical methodUses the variance analysis(ANOVA)of SPSS11.0 software package to carry on the data analysis,during the multi-groups comparisons uses the LSD.Inhibition ratio=(1-experimental group/comparison group)×100%Results1.The normal RAW264.7 under the inverted microscope are adherent cells, to assume the triangle;oval-shaped,the polygon,or synapses.2.Stimulate RAW264.7 cells 48 h with 0.5 uM Tg +50 ug/ml Fuc and 2 uM Tg +50 ug / ml Fuc are able to induce stable apoptosis.When analysis, the difference between normal group(control cells)and apoptosis group is significant,P＜0.05.3.After detected the effect of different concentrations of Huanglianwendan -containing serum(1:10,1:20;1:40,1:80;1:160)to stimulate RAW264.7' proliferation by MTT method,We found1:10 concentrations can promote cell proliferation.Under analysis,the data of experimental group and control group are significant different,p＜0.05.4.Huanglianwendan-containing serum(1:10 final concentration) pre-stimulation RAW264.7 cells can decline the rate of apoptosis,and the data among the serum group,the apoptosis group and the blank Serum control group were significantly different,p＜0.05.5.The Huanglianwendan-containing serum group can not decline SR-A expression.Conclusion0.5μM Tg joint 50μg/ml Fuc can induce the apoptosis of macrophage.Huanglianwendan-containing serum can inhibit apoptosis of RAW264.7. and the data is significant compared both with apoptosis group and the blank serum group.The results showed that the inhibition of macrophage apoptosis may be one of the Huanglianwendan's mechanisms of treating atherosclerosis.Huanglianwendan soup does not have the effect to decline the SR-A.The effect about SR-A declinate the apoptosis would be happened in p38 may or JNK way or both.In addition,the active ingredients of inhibiting apoptosis contains in Huanglianwendan serum may be sensitive to temperatures,and were easy to failure under repeated freeze-thaw.