Qualitative Identification Of The Main Chemical Components Of Xiakuai Xiaoliu Mixture And Its Preliminary Pharmacodynamic Study On NSCLC

Objective:Based on HPLC-TOF/MS analysis platform to established a rapid qualitative identification method for the main chemical components contained in Xiakucao Xiaoliu Mixture.The subcutaneous xenograft model of tumor-bearing mice was used to study the initial pharmacodynamic effect of Xiakucao Xiaoliu Mixture on anti-NSCLC.And explore the possible mechanism of action,in order to provide a theoretical basis for guiding clinical use of drugs.Methods:1.Analysis of clinical application:The inpatients who used the Xiakucao Xiaoliu Mixture in the Department of Oncology,2018.1-2018.6,Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,were randomly selected to analyze the types of cancer applied by Xiakucao Xiaoliu Mixture and the distribution of TCM syndromes of lung cancer patients.2.Experimental part:（1）The main chemical components of Xiakucao Xiaoliu Mixture were analyzed by ACE C18（3.0 mm×150 mm,3.5μm）column;the mobile phase was methanol（A）and 0.1%formic acid water（B）,gradient elution;column temperature 25°C;Spray ion source positive and negative ion mode monitoring,reference ion m/z 121.9856,1033.9881;scanning range m/z 100-1200;using reference materials,mass spectrometry information,comprehensive literature research to establish a database for qualitative identification of each chromatographic peak and herbs Attribution.（2）32 subcutaneous xenograft models of C₅₇BL/6 mice were randomly divided into 4 groups.The rats were given normal saline,cisplatin and Xiakucao Xiaoliu Mixture and concurrently given cisplatin and Xiakucao Xiaoliu Mixture.During the administration,the general condition of the mice was observed,the body weight of the mice was weighed,and the tumor size was measured every day;after 12 days of administration,the tumor was sacrificed,the tumor tissue was removed,and the tumor inhibition rate was calculated;the pathological condition of the tumor tissue was observed by HE staining.Immunohistochemistry was used to detect the expression of Cyclin D1 protein and P16 protein in tumor tissues.Results:1.Analysis of clinical application status:Xiacaocao Xiaoliu Mixture is used in our hospital for the treatment of various cancers,of which lung cancer accounts for more than 90%,and the TCM syndromes for treating lung cancer are Qi and Yin deficiency syndrome.2.Experimental part:（1）Using HPLC-TOF/MS technology to quickly identify 37main chemical components in Xiakucao Xiaoliu Mixture,and administer the main ingredients.（2）Integrating gross observation,body weight and tumor growth,it was found that Xiakucao Xiaoliu Mixture can significantly improve the quality of life of tumor-bearing mice;compared with the model group,Xiakucao Xiaoliu Mixture group,cisplatin group and combination therapy The tumor inhibition rate of the group was15.79%,18.90%,and 50.94%,respectively.The increase rate of the combination group was 1.1599.The pathological observation showed that the tumor tissue of the model group had clear outline,strong growth,and no necrosis.however,the tumor tissue cells of the tumor-bearing mice in each administration group have different degrees of degeneration and necrosis;the immunohistochemistry results show that the tumors of each administration group are small.The expression of P16 protein in murine tumor tissues was higher than that in the model group.The combination group was significantly different from the model group（P<0.05）.Compared with the model group,each drug group significantly downgrade tumor-bearing mice.Expression of Cyclin D1protein in tumor tissues（P<0.05）.Conclusion:Xiakucao Xiaoliu Mixture is used in the treatment of various cancers in our hospital.Most of the lung cancer with qi and yin deficiency,and the curative effect is remarkable.It is found in the clinical treatment of lung cancer.When the combination of the herbicide and the chemotherapeutic drug is safe and reasonable,it can significantly improve the quality of life of patients and improve the clinical treatment effect.Xiakucao Xiaoliu Mixture contains a wide range of chemical components.This study established a HPLC-TOF/MS method for rapid qualitative identification of the main chemical components.The Xiakucao Xiaoliu Mixture can inhibit the growth of NSCLC to some extent.The combination with chemotherapeutic drug cisplatin is more effective;immunohistochemistry results suggest that Xiakucao Xiaoliu Mixture inhibits tumor growth by down-regulating Cyclin D1 protein and up-regulating P16 protein expression.