| ObjectiveTo study the synergistic effect and mechanism of Shashen Maidong Decoction on lung cancer chemotherapy,and provide scientific basis for clinical medication and optimization of lung cancer treatment plan.MethodsAAfter one week of adaptive feeding,48 healthy and clean grade C57 / BL6 mice were randomly divided into four groups according to body weight,namely,the blank group,the lung cancer model group,the cisplatin group and the cisplatin + Shashen Maidong Decoction group,with 12 mice in each group.In addition to the blank group,the three groups were subcutaneously injected with lewis lung cancer cell suspension in the right armpit of mice to establish a mouse lung cancer bearing model.Blank group routine feeding.The model group was given intragastric administration of 0.2 ml sterile water every day and intraperitoneal injection of 0.2 ml sterile water every other day;cisplatin group was given 0.2 ml sterile water by gavage every day and 0.2 ml cisplatin solution by intraperitoneal injection every other day;the cisplatin + Shashen Maidong decoction group was given intragastric administration of0.2 ml Shashen Maidong decoction every day and intraperitoneal injection of 0.2 ml cisplatin every other day.Each group was continuously administered for 14 days.Weight was monitored every other day during feeding.Twenty-four hours after the last administration,the mice were weighed,and the eyeball blood was taken after ether anesthesia to kill the experimental mice.The blood of the orbital vein of the mice was collected,and the thymus tissue,spleen tissue,liver tissue and kidney tissue were separated.The subcutaneous tumor tissue was completely stripped.The subcutaneous tumor tissue,thymus tissue and spleen tissue were placed on the absorbent paper to dry water.After weighing,the spleen index,thymus index and tumor inhibition rate were calculated and the tumor volume was recorded.The changes of white blood cells,platelets and hemoglobin in peripheral blood of mice were detected.The changes of IL-6 and IFN-γ in mouse plasma were detected by ELISA.He staining was used to observe the pathological changes of liver tissue,kidney tissue and tumor tissue.The relative expression of JAK2,STAT1 and STAT3 in tumor tissues was detected by RT-PCR.The protein expression of JAK2,p-JAK2,STAT1,STAT3,p-STAT3,Bcl-2 and PIM1 was detected by Western Blot.Result1.Study on the attenuated effect of Shashen Maidong decoction on lung cancer chemotherapy（1）General situation : The mice in the blank group had no abnormal living conditions and the highest body weight;lung cancer model group mice hair significantly withered,slow activity,stress response weakened,the lowest weight;compared with lung cancer model group,the general condition of mice in cisplatin group was improved,the hair luster was improved,the activity was increased,the stress response was enhanced,and the weight of mice in cisplatin group was higher than that in lung cancer model group.Compared with lung cancer model group and cisplatin group,the survival status of mice in cisplatin + Shashen Maidong decoction group was significantly improved,the activity was faster,the stress response was more sensitive,and the body weight was higher than that of lung cancer model group and cisplatin group.（2）Immune organ index : thymus index and spleen index of blank group were significantly higher than those of lung cancer model group（P < 0.01）;compared with lung cancer model group,the spleen index and thymus index of cisplatin group were decreased（P < 0.05）.Compared with lung cancer model group and cisplatin group,the spleen index and thymus index of cisplatin + Shashen Maidong decoction group were increased（P < 0.01）.（3）Changes of white blood cells,platelets and hemoglobin in peripheral blood : white blood cells,hemoglobin and platelets in peripheral blood of blank group were higher than those of lung cancer model group（P < 0.01）;compared with lung cancer model group,white blood cells,hemoglobin and platelet in cisplatin group were decreased（P < 0.05）.Compared with lung cancer model group and cisplatin group,the white blood cell,hemoglobin and platelet in cisplatin group + Shashen Maidong decoction group were increased（P < 0.05）;（4）He staining :He staining of liver tissue : There was no obvious pathological change in liver tissue of mice in blank group and lung cancer model group;hepatocytes in cisplatin group showed swelling,loose and disordered arrangement,and focal necrosis area increased significantly.The structure of liver cells in cisplatin + Shashen Maidong decoction group was relatively complete and clear,and the arrangement was relatively neat.Necrotic cells were scattered around the portal area,and the damage was mild.He staining of renal tissue : In the blank group and lung cancer model group,renal tubules and glomeruli were normal and complete,and the surrounding cells were arranged in neat rows,without obvious abnormal changes.Compared with the blank group and lung cancer model group,the kidney tissue of mice in cisplatin group showed obvious renal tubular epithelial cell lesions,interstitial space was significantly increased,cell arrangement was loose and disordered,and a large number of inflammatory cell infiltration and cell shedding and necrosis were observed,showing obvious pathological changes.Compared with the cisplatin group,the injury of renal tubular epithelial cells in the cisplatin + Shashen Maidong Decoction group was significantly reduced,the interstitial space was significantly reduced,and the inflammatory cell infiltration was also significantly improved.2.Study on the synergistic effect and mechanism of Shashen Maidong decoction on lung cancer chemotherapy（1）Tumor growth curve,image recording and tumor inhibition rate : The growth rate and tumor volume of lung cancer model group were much higher than those of cisplatin group and cisplatin +Shashen Maidong Decoction group,and the growth rate and volume of cisplatin group were higher than those of cisplatin + Shashen Maidong Decoction group.The image records also showed that the tumor volume of mice in the lung cancer model group was significantly larger than that in the cisplatin group and the cisplatin + sashen maidong decoction group,while the tumor volume of mice in the cisplatin group was significantly larger than that in the cisplatin + sashen maidong decoction group.The inhibitory rate of cisplatin group was significantly lower than that of cisplatin + Shashen Maidong decoction group（P < 0.01）.（2）He staining of tumor tissue : In the lung cancer model group,the tumor cells were densely arranged,the infiltration of immune cells was not obvious,the division phenomenon was more,there was no necrosis and nuclear staining.In cisplatin group,there was a slightly large necrotic area,tumor cells arranged loosely and irregularly,vacuolization and necrosis,nuclear division decreased,and immune cell infiltration increased.The tumor tissue of mice in cisplatin + Shashen Maidong decoction group showed a large area of necrosis,tumor cells arranged loosely and irregularly,the phenomenon of nuclear division was significantly reduced,and the infiltration of immune cells was the most obvious..（3）The changes of JAK2,STAT1 and STAT3 gene expression in tumor tissues : compared with the lung cancer model group,the JAK2 and STAT3 gene expression in tumor tissues of mice in cisplatin group and cisplatin + Shashen Maidong Decoction group were significantly decreased（P < 0.01）,and the STAT1 gene expression was significantly increased（P < 0.01）.Compared with the cisplatin group,the JAK2 and STAT3 gene expressions in the tumor tissues of mice in the cisplatin + Shashen Maidong Decoction group were significantly down-regulated（P < 0.01）,and the STAT1 gene expression was significantly up-regulated（P < 0.01）..（4）The protein expression changes of JAK2,p-JAK2,STAT1,p-STAT3,Bcl-2 and PIM1 in tumor tissues : compared with the lung cancer model group,the protein expressions of JAK2,p-JAK2,STAT3,p-STAT3,Bcl-2 and PIM1 in the tumor tissues of mice in cisplatin group and cisplatin + Shashen Maidong Decoction group were significantly decreased（P < 0.01）,and the protein expression of STAT1 was increased.Compared with cisplatin group,the expression of JAK2,p-JAK2,STAT3,p-STAT3,Bcl-2 and PIM1 protein in tumor tissue of cisplatin + Shashen Maidong Decoction group was significantly decreased（P < 0.01）,and the expression of STAT1 protein was up-regulated..（5）Changes of plasma IL-6 and IFN-γ : Compared with the normal group,the levels of plasma IL-6 in lung cancer model group,cisplatin group and cisplatin + Shashen Maidong Decoction group were significantly increased,and the levels of IFN-γ were significantly decreased.Compared with the lung cancer model group,the content of IFN-γ in cisplatin group and cisplatin + Shashen Maidong Decoction group was significantly increased（P < 0.01）,and the content of IL-6 was significantly decreased（P < 0.01）.Compared with cisplatin group,the content of IFN-γ in plasma of cisplatin + Shashen Maidong Decoction group was significantly increased（P < 0.01）,and the content of IL-6 was significantly decreased（P < 0.01）.Conclusion1.Shashen Maidong Decoction has a attenuated effect on lung cancer chemotherapy.It can improve the quality of life of mice,increase the spleen index and thymus index,increase the content of white blood cells,platelets and hemoglobin in peripheral blood,and improve the pathological damage of liver tissue and kidney tissue.2.Shashen Maidong Decoction has synergistic effect on chemotherapy of lung cancer.It is mainly manifested in inhibiting the growth of tumor tissue,reducing tumor volume,improving tumor inhibition rate,promoting tumor necrosis,increasing plasma IFN-γ and decreasing plasma IL-6 content,reducing JAK2,STAT3 and increasing STAT1 gene expression in tumor tissue,reducing JAK2,p-JAK2,STAT3,p-STAT3,Bcl-2 and PIM1 protein expression in tumor tissue,and increasing STAT1 protein expression.Its mechanism may be related to inhibiting JAK / STAT signaling pathway. |
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