| Purpose: To study the effect of modified Chaishao Liujunzi decoction on the expression of SRC protein in human orthotopic Pancreatic cancer(PC)tissue implanted subcutaneously in nude mice.To investigate the mechanism of modified Chaishao Liujunzi decoction on pancreatic cancer,and to provide basic theoretical basis for the use of modified Chaishao Liujunzi decoction in the clinical treatment of pancreatic cancer.Material and method:The nude mice were subjected to chronic restraint by clamping their tails with plastic clips for about 30 minutes every day to establish the nude mice model of liver stagnation and spleen deficiency induced by chronic restraint.At the same time,the nude mice were treated with 5% vinegar solution for drinking water throughout the day to establish the nude mice model of liver stagnation and spleen deficiency.Human pancreatic cancer Bx PC-3 cells were inoculated subcutaneously into the armpit of the right forearm of mice with liver stagnation and spleen deficiency.After successful modeling,24 mice with liver stagnation and spleen deficiency were divided into 5 groups by random number table method: model control group(0.9% sodium chloride solution),low dose of traditional Chinese medicine group(modified Chaishao Liujunzi decoction concentration 0.473g/ml;0.2ml),Chinese medicine medium dose group(modified Chaishao Liujunzi Decoction concentration 0.945g/ml,0.2ml),western medicine group(dasatinib0.2mg/ mouse),and combination group(modified Chaishao Liujunzi decoction concentration 0.945g/ml,0.2mg+ dasatinib 0.2mg/ mouse).The size of subcutaneously implanted tumors in each nude mouse was measured every 4 days using a vernier caliper.After 20 days of administration,the mice were sacrificed,and the transplanted tumors were removed.After observation,the tumors were fixed with paraformaldehyde and frozen at-80℃ for later use.The body weight of the nude mice before and after treatment was weighed by electronic scale,and the tumor weight of the implanted tumor was weighed after sampling,and the tumor inhibition rate was calculated.The data were used to analyze the transplanted tumor volume at 0,4,8,12,16,and 20 days after administration.Immunohistochemical staining and Western blot WB were used to detect the expression level of SRC protein in the transplanted tumor tissue of pancreatic cancer in nude mice.SPSS 26 statistical software was used to analyze the data obtained during the experiment,and the conclusions were drawn.0.05 was considered statistically significant.Results:1.Tumor formation rate: the model was successfully established when the human in situ cancer cell Bx PC-3 cell suspension was visible to the naked eye and became larger day by day,and the maximum diameter of the tumor was ≥5mm.Twenty-four nude mice were successfully established.2.General status of nude mice: mice were listless,less active,lethargic,lazy,arched back curled up,nude mice lost weight significantly,loose stool,and some nude mice showed ulceration and scabs at the tumor sites as the tumor grew larger.3.The weight and tumor volume of nude mice in each group before and after treatment: compared with the model group,the average weight loss trend of nude mice in each group was significantly reduced,and the tumor volume was also significantly reduced(P<0.05).After treatment,the average weight of nude mice in each group was: model control group(15.08±0.29)g,Chinese medicine low-dose group(15.72±0.29)g,Chinese medicine medium-dose group(16.64±0.29)g,western medicine group(17.58±0.42)g,combined treatment group(18.32±0.32)g.The tumor volumes were: Model control group:(1069.203±52.303)mm3,Chinese medicine low-dose group:(1033.585±32.944)mm3,Chinese medicine medium-dose group:(949.190±13.265)mm3,western medicine group:(813.542±31.803)mm3;(760.006±18.832)mm3 in combination group.Compared with the model group,the growth of the implanted tumors was inhibited and the tumor volume was significantly reduced in the treated groups(P<0.05).4.Calculation of tumor weight and tumor inhibition rate: at the end of gavage,the tumor weight of nude mice in each group was weighed as follows:(2.04±0.04)g in the model control group,(1.91±0.04)g in the low dose group,(1.83±0.04)g in the middle dose group,(1.53±0.05)g in the western medicine group,and(1.41±0.01)g in the combination group.The results showed that the tumor weight of the treated groups was significantly smaller than that of the model control group(P<0.05),and the combination group had the lightest tumor weight and the best curative effect.The tumor inhibition rate was 6.3% in the low dose group,10.2% in the middle dose group,25% in the western medicine group,and 30.9% in the combination group.5.Protein expression: Immunohistochemical staining and Western blot(WB)showed that the expression of SRC protein in the middle dose group,the western medicine group and the combination group was significantly lower than that in the model control group(P<0.05).Compared with the western medicine group and the combination group,the low dose group of traditional Chinese medicine had statistical significance(P<0.05).Compared with the western medicine group and the combination group,the Chinese medicine medium dose group had statistical significance(P<The results of Western blot showed that compared with the model control group,the expression level of SRC protein in the tumor tissue of nude mice in the low-dose TCM group was significantly decreased(P<0.05).However,the results of immunohistochemical staining showed that there was no statistically significant difference between the low dose group and the model control group(P >0.05),which was analyzed as the effect of low dose of traditional Chinese medicine caused unstable results.Conclusion:1.In the nude mouse model of pancreatic cancer,the tumor volume and weight of the treated groups were smaller than those of the model control group,and the weight loss trend of the treated groups was also slower than that of the model control group,indicating that both modified Chaishao Liujunzi decoction and dasatinib had anti-tumor proliferation and growth effects.In addition,the tumor inhibition effect of the combined drug group was the most obvious,indicating that the combination of modified Chaishao Liujunzi decoction and dasatinib has an enhanced effect.In the experiment,the therapeutic effect of the middle dose group of Chinese medicine was better than that of the low dose group of Chinese medicine,which confirmed the dosage commonly used in clinical practice.After treatment,the body weight of the nude mice in the combination group was the highest,indicating that the drug treatment can reduce the cachexia of pancreatic cancer to a certain extent.2.The results of immunohistochemical staining and Western blot showed that Jiawei Chaishao Liujunzi Decoction could inhibit the expression of SRC protein in pancreatic cancer tissues,thereby inhibiting the proliferation and growth of pancreatic cancer tissues. |
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