Effect Of Shengxian Decoction On The Proliferation Of Human Lung Adenocarcinoma Cell A549

[Objective]The in vitro cell experiments were used to demonstrate the effect of different dosage groups of Sheng Xian Decoction on the growth and proliferation of human lung adenocarcinoma A549 cells,and a nude mouse model of human lung adenocarcinoma A549 transplanted tumor was selected as the research object.The dose-effect relationship of the dose of medicine on the inhibition of lung adenocarcinoma growth and some biological factors related to its effect,so as to broaden the clinical application scope of Sheng Xian Decoction and provide experimental basis for the traditional Chinese medicine compound to interfere with lung adenocarcinoma.[Method]1.Study on the intervention of Sheng Xian decoction-containing serum in the proliferation of human lung adenocarcinoma cell A549: selecting A549 lung adenocarcinoma cell line,culturing human lung adenocarcinoma A549 cell line,and using serum pharmacology to prepare SD rat drug-containing serum The cell growth and proliferation of each group were detected by MTT method.2.Study on the intervention of Shengxian Decoction in the growth of lung adenocarcinoma A549 nude mice transplanted tumors:0.2ml human lung adenocarcinoma A549 cell suspension was inoculated subcutaneously in the right armpit near the dorsal part of the nude mice at a concentration of 1x106 / ml.A549 nude mice xenograft model.Observe the growth and proliferation of tumor cells.When the tumor is visible to the naked eye,the modeling is successful.The validity of the model was evaluated by observing histomorphology through pathological sections.Twenty-five nude mice successfully modeled were randomly divided into 5 groups,5in each group.The groups were: blank control group(MG),cisplatin(DDP)group,high-dose soup group(SXT-H),middle-dose soup group(SXT-M),low-dose soup group(SXT)-L).It was observed that when the average diameter of the subcutaneousmass reached about 5 mm,administration was started immediately.Tumors were dissected out after 21 days of continuous administration.The weight change curve and the growth curve of the transplanted tumor in the nude mice were drawn,and the inhibition rate of the drugs on the transplanted tumor in each group was calculated.The pathology of the transplanted tumor in the nude mice was observed by HE staining.3.Shengxian Decoction intervention in lung and spleen tissues of nude mice with lung adenocarcinoma xenografts and its relationship with regulating serum TNF-α and HIF-1α: The above model was used to take the lung and spleen tissues of nude mice for 21 days to make paraffin Sections were used to observe the pathological conditions of lung and spleen by HE staining;Elisa was used to detect the TNF-α and HIF-1α in the serum of nude mice,and to explore the biological effects of Sheng Xian Decoction on lung adenocarcinoma.[Results](1)Detection by MTT method showed that the Sheng Xian decoction-containing serum significantly inhibited the proliferation of human lung cancer cell A549,and the high-,medium-,and low-dose groups of the drug-containing serum had effects on human lung adenocarcinoma A549 cells for 24 h,48h,72 h Proliferation inhibition rate was obtained,that is,the inhibition rates of 24 h SXT-H,SXT-M,and SXT-L A549 cells were 16.37%,13.61%,and 5.35%;48h inhibition rates were 27.23%,24.44%,and 15.94,respectively.%;72h inhibition rates were 35.01%,30.98%,and 24.69%,respectively.Therefore,the inhibition rate of Sheng Xian Decoction-containing serum on lung adenocarcinoma cell proliferation was positively correlated with time and dose,that is,the higher the concentration,the higher the inhibition rate over time.(2)A nude mouse model of human lung adenocarcinoma A549 xenograft was successfully constructed with a tumor formation rate of 100%.Lung adenocarcinoma transplanted tumors are located in the subcutaneous area of ??the right axilla near the dorsal part of the nude mice.They can rub against millet granular protrusions.The transplanted tumors have small nodules visible to the naked eye.The texture is hard.,Poor mobility,complete envelope,and a small part will have adhesions.It was observed that the tumors were fish-like red and hard in texture.The HE staining of the transplanted tumor tissue and the HE staining of the transplanted tumor tissue showed that the tumor cells grew vigorously and the tumor cells were densely arranged.Some cytoplasm contained circular lipid droplets,and multiple basophilic exudation sites were seen between the cells.The nucleus varies in size,with a small atypia,obvious nucleoli,pathological mitotic phase,and no obvious area of ??degeneration and necrosis.By inhibiting the proliferation of A549 nude mice at high,medium and low doses of Sheng Xian Decoction,the inhibition rate was obtained.It can be observed in the table that with time,the inhibition rate of cisplatin on A549 nude mice transplantation tumors gradually increased,and The tumor rates were 9.17%,23.15%,35.48%,38.63%,42.47%,53.57%,and 57.36%;compared with the cisplatin group,the high depression and high and low dose groups also gradually inhibited the rate over time.Increased,the inhibition rate of cisplatin group on A549 nude mice transplantation tumors was weaker at the same time;among the various dose groups of Sheng Xian Decoction,the inhibitory effects of Sheng Xian Decoction on A549 nude mice were better than that of Sheng Xian Decoction The low-dose group of Shengtang was higher than the low-dose group of Shengkentang.The results showed that Shengzhan Decoction had inhibitory effect on lung cancer A549 nude mice xenografts,and the inhibition rate was positively correlated with time.That is to say,the higher the inhibition rate over time,the higher the dose of Shengzhan Decoction was better than other dose groups.(3)Sheng Xian Decoctioninterfered with lung adenocarcinoma xenografts in nude mice.No tumor was found in HE staining of the lung and spleen tissues.Compared with the model group,cisplatin group,Shengxue Tang high and medium dose groups were transplanted in the serum of nude mice.The level of HIF-1α was significantly reduced,which was statistically significant(P <0.01).There was a downward trend in the low-dose group of Shengkatang,but it was not statistically significant.Compared with the cisplatin group,the high-dose soup group and the low-dose group The serum levels of HIF-1α in nude mice with A549 transplantedtumors were significantly different,but there was no significant difference in the middle-dose group of Sheng Xian Decoction.The results showed that the high-,low-,and medium-dose groups of Sheng Xian Decoction had inhibitory effects on HIF-1α in nude mice with A549 tumors,and the dose-inhibitory effect in Sheng Xian Decoction was obvious.Compared with the model group,serum TNF-α in nude mice with A549 xenograft tumors in the cisplatin group,Sheng Xian Decoction high and middle dose groups was significantly reduced,which was statistically significant(P<0.01 or P <0.05).There was a downward trend in the group,but there was no statistical significance.Compared with the cisplatin group,the serum TNF-α levels of A549 xenograft nude mice in the high-dose and low-dose groups were significantly different from those in the high-dose and low-dose groups.difference.The results showed that the high-,medium-and low-dose groups of Sheng Xian Decoction had inhibitory effects on serum TNF-α in nude mice with A549 transplanted tumors.[Conclusion]Sheng Xian Decoction can effectively inhibit the growth and proliferation of human lung adenocarcinoma cell A549,and its effect may be to reduce the level of TNF-α and HIF-1α in the serum of model nude mice,affect the hypoxic microenvironment of lung adenocarcinoma,and induce tumor cell necrosis.It may be one of the mechanisms that delay the development of lung adenocarcinoma.