Luoxiaoyin Induces Lymphadenoma Raji Cell's Apoptosis Preliminary Experimental Study

Objective:Malignant lymphoma is a solid tumors origin from lymphoid hematopoietic tissue.In our country,although malignant lymphoma are relatively rare,in recent years the munber of new cases increasde year by year.Domestic and foreign research for lymphoma has been over a hundred years.Modern medicine in the treatment of lymphoma has been made progress,but the progress,but the prognosis remains poor.Although Chinese medicine literature did not have the disease of malignant lymphoma,based on its clinical symptoms and signs,malignant lymphoma belong to the course of Chinese medicine "luoli","shirong","shiju","ehe","tanhe",etc.So as to,there is a lot of valuable experience worth learning in documented for treat lymphoma .My instructor Dr.Liuyaxian referenceliterture(suchas:"yixuexinwu","yixuezhongzhongcanxilu","shanghanwenyitiaobian",etc.),and taking into account the clinical manifestations of lymphoma and morbidity characteristics,combined with clinical treament,presceibe Luoxiaoyin.Many years of clinical practice for patients with malignant lymphoma has proved Luoxiaoyin obtain a better therapeutic effect in reducing radiotherapy and chemotherapy toxicity,relieving clincal symptoms,improving quality of life,extend the survival time.And radiotherapy and chemotherapy for the prevention of intermittent period of lymphoma recurrence and metastasis have a significant effect. Of not,should not radiotherapy and chemotherapy patients,as well as the advanced treatment of patients must also have the effect.Some patients can even be long-term surviva with malignant lymphoma.Basis on the therapeutic efficacy,we conducted this experimental study alimed at exploring Luoxiaoyin at induced Burkitt's lymphoma cell line Raji cell apoptosis in the role of the mechanism of the treatment of malignant lymphoma of the parties,to provide the objective basis,further guidance and to expand clincal application.Methods:Application of in vitro cell culture technology,based on the consult of the literature and preliminary experiment,selected 0.5mg/ml,1.0mg/ml,2.0mg/ml,4.0mg/ml,8.0mg/ml five final concentration,and set RPMI-1640 for the control group,cultured with tumor cells 24,48,72 hours later,to assay scrofula eliminate water solution on the role of tumor cells in vitro after the cell survival rate by trypan blue exclusion staining.Use MTT to assay Luoxiaoyin's inhibition of lymphadenoma Raji cell proliferation. Use Flow cytometry to detect the apoptosis rate and cell cycle distribution after different concentrations of Luoxiaoyin water solution effect-ed in the tumor cells 48h.Use Flow cytometry detect the role of apoptosis suppressor gene bcl-2 protein expression after different concentrations of Chinese medicine effected the tumor cell.Results:1 Trypan blue exclusion staining results showed that after 1.0mg/ml,2.0mg/ml,4.0mg/ml, 8.0mg/ml concentration Luoxiaoyin effected Raji cells 24,48,72 hours,cell growth were significantly inhibited,compared to the control group there is significant difference (P<0.05),in the concentration range in a dose-time-dependent manner.However,concentration of 0.5mg/ml group compared to the control group no significant difference (P>0.05).Various traditional Chinese medicines density group of comparison difference is obvious(P<0.05).2 MTT colorimetric assay results showed that after 1.0mg/ml,2.0mg/ml,4.0mg/ml, 8.0mg/ml concentration Luoxiaoyin effected Raji cells 24,48,72 hours,various densities group can suppress the tumour cell multiplication,compared to the control group there is significant difference (P <0.05),and growth inhibition rate and drug action concentration and time was positively correlated in a dose-time-dependent manner. However,concentration of 0.5mg/ml group compared to the control group no significant difference (P >0.05).3 Use Flow cytometry to detect cell cycle distribution and apoptosis rate showed that after 1.0mg/ml,2.0mg/ml,4.0mg/ml, 8.0mg/ml concentration Luoxiaoyin effected Raji cells 72 hours,G0/G1 phase cells increased,S phase cell's the proportion descreased,compared to the control group there is significant difference (P<0.01);while the concentration of 0.5mg/ml group,although to make the proportion of G0/G1 phase cells increased,S phase cell ratio decreased,compared to the control group no significant difference (P>0.05).After 2.0mg/ml,4.0mg/ml,8.0mg/ml concentration Luoxiaoyin effect tumor Raji cells 72 hours,cell weak perishing rate markup,compared to the control group there is significant difference (P<0.05).After 0.5mg/ml,1.0mg/ml concentration Luoxiaoyin effected tumor cell 72 hours,compared to the control group no singnificant differences (P>0.05).4 The expression of bcl-2 protein detected by Flow cytometry showed that bcl-2 protein expression was significantly reducec,followed the increased concentration of Chinese medicine.1.0mg/ml,2.0mg/ml,4.0mg/ml,8.0mg/ml concentration group compared with the control group had significant difference (P<0.01). However,concentration of 0.5mg/ml group compared to the control group no significant difference (P>0.05).Various traditional Chinese medicines density group of comparison difference is obvious(P<0.01).Conclusion:Through the cells in vitro experimental's results can be found that Luoxiaoyin in a certain range of concentration can obviously inhibit malignant lymphoma Raji cell proliferation dose-and time-dependent.Luoxiaoyin can affect cell cycle distribution,cause G0/G1 phase arrest,induce tumor cell apoptosis.Luoxiaoyin's these roles may be related to it can reduce bcl-2 protein's expression.