Experiment Research Of Inhibiting Effect Of BuShenShuGan Formula On Lung Adenocarcinoma A549Cells And Lewis Lung Cancer Mice

Objective:To investgate the effect of Bushen Shugan(BSSG) formula on immune function and endocrine system of lewis lung cancer mice and the effect of serum containing BSSG on the proliferation of A549cells and discuss the possible underlying mechanism.Methods:In Vitro Experiment.30healthy SD mice were randomly divided into five groups:model control group、DDP group、low-dose group、middle-dose group、 high-dose group of serum containing BSSG. Model control group was given saline4ml by intragastric administration every day; DDP group was given cisplatin solution2ml(containing DDP1mg)by the way of intraperitoneal injection on the first day、the third day and the fifth day; low-dose group was given BuShenShuGan formula4ml/d (containing crude chinese herb7g) by intragastric administration; middle-dose group was given BuShenShuGan formula4ml/d (containing crude chinese herb14g) by intragastric administration; high-dose group was given BuShenShuGan formula4ml/d (containing crude chinese herb21g) by intragastric administration; all the treaments lasted5days.At last,the mice blood were collected by puncturing the aorta abdominalis2hours after treaments on the fifth day.The serum containing BSSG was made,and store in the refrigerator of-20℃.We can observe the effect of serum containing BSSG on the proliferation of A549cells.In Vivo Experiment.40C57BL/6mice that had been successfully transplanted Lewis lung cancer model were randomly divided into four groups:model control group、DDP group、BSSG group、BSSG combined with DDP group. Model control group was given saline0.4ml by intragastric administration every day, and the same time was given saline0.5ml by the way of intraperitoneal injection on the first day、 the third day and the fifth day;DDP group was given cisplatin solution0.5ml (containing DDP0.1mg) by the way of intraperitoneal injection on the first day、the third day and the fifth day,and the same time was given saline0.4ml by intragastric administration every day; BSSG group was given BuShenShuGan formula0.4ml/d (containing crude chinese herb0.7g) by intragastric administration, and the same time was given saline0.5ml by the way of intraperitoneal injection on the first day、 the third day and the fifth day; BSSG combined with DDP group was given BuShenShuGan formula by intragastric administration and cisplatin solution by intraperitoneal injection which was the same as BSSG group and DDP group;all the treaments lasted14days. We observed the mental state、diet and activities of mice everyday, and measured body weight and tumor volume of mice every two days. At last,all the mice blood were collected by taking off the eyeballs on the fifteenth day, and tumor weight and numbers of spontaneous lung metastasis were also measured.The concentration of serum IL-2、IL-10、E2and T were measured by ELISA.Results:1、In vitro experiment, compared with model control group, low-dose group of BSSG(48h、72h)、middle-dose group、high-dose group and DDP group can inhibit the proliferation of A549cells(p<0.01),which has the dose effect and time effect. It is almost no difference between high-dose group and DDP group (p>0.05)2、In Vivo Experiment.Compared with model control group, the other treatment group could significantly inhibit tumor growth, and the difference was significant (p<0.01). Compared with BSSG group, the average tumor weight of BSSG combined with DDP group decreased significantly (p＜O.05).3、Compared with model control group, the other treatment group could significantly inhibit the spontaneous lung metastasis (p<0.05or p<0.01).4、After the experiment finished, compared with model control group, the weight of DDP group decreased (p<0.05); BSSG group and BSSG combined with DDP group were significantly increased (p<0.05), there was no difference between Chinese herbal medicine group and combined group (p>0.05). Compared with model control group, tumor volume of every treatment group was significantly smaller (p<0.05); compared with DDP group and BSSG group, though the tumor volume of combined group was smaller, the difference was not significantly(p>0.05).5、Compared with model group, the blood serum IL-2of all treatment groups increased significantly (p<0.01); compared with DDP group, BSSG combined with DDP group increased significantly (p<0.01). Compared with model control group, the blood serum IL-10of the treatment groups significantly decreased (p<0.01); compared with DDP group, the blood serum IL-10of BSSG group was significantly decreased (p<0.05).6、Compared with model control group, the blood serum E2of all treatment groups decreased significantly (p<0.01); compared with DDP group and BSSG group, the blood serum E2of BSSG combined with DDP group decreased significantly (p<0.01). Compared with model control group, the blood serum T of all treatment groups increased significantly (p<0.01); compared with DDP group and BSSG group, the blood serum T of BSSG combined with DDP group increased significantly (p<0.01). Compared with model control group, the blood serum E2/T of all treatment groups decreased significantly (p<0.01); compared with DDP group and BSSG group, the blood serum E2/T of BSSG combined with DDP group decreased significantly (p<0.01).Conclusion:1、Serum containing BSSG can inhibit the proliferation of A549cells, which has the dose effect and time effect.2、BuShenShuGan Formula could significantly inhibit growth and spontaneous lung metastasis of Lewis lung cancer, and there was synergistic effect with cisplatin.3、BSSG Formula can make an effect on the level of blood serum IL-2、IL-10、E2and T.It can improve the functions of immunity and regulate internal secretion, and this may be one of the mechanisms of the inhibition of tumor growth.4、BuShenShuGan Formula could improve the quality of the mice life, reduce the side effects caused by chemotherapy such as weight loss and so on.