| Objective:Through clinical research,in vivo and in vitro experimental studies to investigate the mechanism of Jianpi Yangzheng Xiaozheng Recipe treating gastric cancer,the effects of Jianpi Yangzheng Xiaozheng Recipe on the aerobic glycolysis level of gastric cancer and the gastric cancer sternness were exploredMethods:1.All advanced gastric cancer cases originated from Jiangsu Hospital of Traditional Chinese Medicine,Affiliated Hospital of Nanjing University of Traditional Chinese Medicine.From February 2019 to February 2020,outpatients and inpatients of the Oncology Department of Jiangsu Provincial Hospital of Traditional Chinese Medicine.Divided 4 into experimental group(Jianpi Yangzheng Xiaozheng Recipe+Chemotherapy)and control group(chemotherapy).The two groups were treated continuously for 6 weeks.The patients in the above cases were drawn blood before and after treatment,using a lactic acid detection kit and a lactic dehydrogenase detection reagent.The kit measures the blood lactate(LD)and lactate dehydrogenase(LDH)levels before and after treatment in the two groups of patients.2.The MTT method was used to observe the effects of Jianpi Yangzheng Xiaozheng Recipe and cisplatin on the proliferation of gastric cancer HGC-27 cells.(2mg/ml,4mg/ml,8mg/ml)on the colony-forming ability of gastric cancer HGC-27 cells;AnnexinV/PI double staining method was used to study the effect of Jianpi Yangzheng Xiaozheng recipe on gastric cancer cell apoptosis.3.Glucose detection kit and lactic acid detection kit were used to detect the lactate production rate and glucose consumption rate of gastric cancer cells by Jianpi Yangzheng Xiaozheng Recipe;Western blot was used to detect the expression of key enzymes of the aerobic glycolytic metabolic pathway in gastric cancer cells after administration.4.Subcutaneous injection of gastric cancer HGC-27 cells to establish a subcutaneous xenograft tumor model in nude mice,and observe the size of subcutaneous xenograft tumors in nude mice,weight changes in nude mice,and the lactate dehydrogenase test kit and lactate test kit to detect health Blood lactate dehydrogenase and lactic acid levels in nude mice and nude mice in each group.5.Jianpi Yangzheng Xiaozheng Recipe was detected and analyzed by flow cytometry and cell pelleting test Effects on gastric cancer stem cell-related markers and cell sphericity;Western blot was used to detect the expression of stem cell-associated proteins in gastric cancer cells after administration.6.Western blot was used to detect the expression of PI3K/AKT/mTOR signaling pathway-related proteins in gastric cancer cells after administration.Use IGF-1 as a PI3K/AKT/mTOR signaling pathway agonist to detect the effects of IGF-1 alone or in combination with Jianpi Yangzheng Xiaozheng Recipe on the lactate production rate,glucose consumption rate,and cell spheroidizing ability of gastric cancer cells.Blots were used to detect changes in stemness-related proteins,key enzymes of the aerobic glycolytic metabolic pathway,and PI3K/AKT/mTOR signaling pathway-related proteins after administration.Result:1·The use of Chinese medicine on the basis of chemotherapy can significantly increase the kps score of patients with advanced gastric cancer,and the increase in kps score after treatment in the chemotherapy alone group is not statistically significant,suggesting that Jianpi Yangzheng Xiaozheng Recipe can improve the life of patients during treatment Significant quality effect;2.From the perspective of treatment effect,the effective rate of the traditional Chinese medicine combined with chemotherapy group is not statistically significant;3.Serum aerobic glycolysis indicators lactate dehydrogenase and lactic acid levels in patients with advanced gastric cancer are higher than normal people,which indicates that their metabolic mode is biased towards aerobic glycolysis,and from the test results after treatment,only use Chemotherapy drugs increase the level of aerobic glycolysis in patients,and in the case of combined use,the relevant indicators in patients are reduced,and the difference is statistically significant,suggesting that Jianpi Yangzheng Xiaozheng recipe can reduce aerobic conditions in patients with advanced gastric cancer.Glycolysis level.4.Jianpi Yangzheng Xiaozheng Fang water extract(JPYZXZF)can inhibit the viability of gastric cancer HGC-27 cell line in vitro,induce apoptosis,reduce the colony formation rate,and is concentration-dependent;low-dose JPYZXZF(1/3 IC50)When used in combination with the chemotherapy drugDDP,it can play a role in increasing the sensitivity of DDP chemotherapy;5.JPYZXZF can reduce the glycolysis level of gastric cancer cell HGC-27 tumors and inhibit the expression of key enzymes during aerobic glycolysis;6.Jianpi Yangzheng Xiaozheng Recipe can inhibit the subcutaneous transplantation of nude mice bearing gastric cancer,and can improve the weight loss caused by tumors and improve their quality of life;Compared with healthy nude mice,the aerobic glycolysis level in nude mice bearing gastric cancer is enhanced,and the Jianpi Yangzheng Xiaozheng recipe can inhibit the glycolytic level in nude mice bearing gastric cancer.7.JPYZXZF can down-regulate the ratio of CD44+CD24+ALDH+cell subsets of gastric cancer HGC-27 cells,reduce the ability of cells to spheroidize in vitro,and down-regulate the expression of sternness-related transcription factors,suggesting that JPYZXZF can inhibit the dryness of gastric cancer cells.8.The up-regulation of the PI3K/AKT/mTOR signaling pathway can increase the level of aerobic glycolysis of gastric cancer cells.At the same time,the sternness of gastric cancer cells also increases,and JPYZXZF can save it and increase the rate of aerobic glycolysis.Malignant effect,while reverting the inhibition of gastric cancer sternness;conclusion:1.Compared with healthy people,patients with advanced gastric cancer have a glucose metabolism pattern that favors aerobic glycolysis;Jianpi Yangzheng Xiaozheng Decoction can reduce aerobic glycolysis in patients with advanced gastric cancer.2.Jianpi Yangzheng Xiaozheng Recipe can induce apoptosis of gastric cancer cells,increase the sensitivity of gastric cancer cells to chemotherapy,and its mechanism of action may be related to the down-regulation of aerobic glycolysis and dryness of gastric cancer cells.3.Jianpi Yangzheng Xiaozheng Recipe can down-regulate the expression of key enzymes in the downstream aerobic glycolysis pathway by inhibiting the cascade of PI3K/AKT/mTOR/HIF1-α/c-Myc,and reduce the level of aerobic glycolysis.Thus inhibiting the dryness of gastric cancer cells. |
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