Material Basis Screening System Of Spica Prunellea On Prevention And Therapy Of Lung Cancer

The thought of "following the prognosis of a disease" in Chinese medicine has concordance with modern tumor precaution view.The "following the prognosis of a disease" includes:prophylaxis before disease,prevention pathological changes since has gotten a disease,prevention of relapse after recovery.Tertiary prevention of modern phymatology includes:primary prevention means interrupt or delay the disease before tumor occurrence; secondary prevention means therapy intervention in the early period of tumor or deuto-clinical period,and sublime the recovery rate;third prevention means anti-relapse, anti-sequelae and anti-metastasis.With the guidance of the "following the prognosis of a disease" thought in Chinese medicine,this topic research takes the spica prunellea as the model medicine for the introduction,digestion and innovation of the screening system,including in vitro effect screening system,the in vivo biochemistry animal model screening system in advance,establishes material basis screening system of Chinese medicine on the prevention of lung cancer.Further to explain the material basis for the prevention and therapy of lung cancer based on the screening system.The main contents containing in the dissertation are as follows:1.Establishment of the material basis screening system of spica prunellea on the prevention and therapy of lung cancer①The biochemistry pre-selection system:Establishment of antioxidation models:To establish the antioxidation models,take the SOD,MDA as the investigation targets to evaluate the oxidation resistance mechanism of spica prunellea in vivo.Results showed P.Hispida Bent from Sichuan had the strongest antioxidation ability(SOD was 52.39±2.41 U/mL and MDA was 3.20±1.25 nmol/mL),SOD and MDA of P.asiatica Nakai from Jiangsu were 47.03±2.84 U/mL, 3.57±0.98 nmol/m,respectively;SOD and MDA of Prunella vulgaris L.from Anhui were 3.23±0.67 nmol/mL and 48.28±1.85U/mL respectively.Spica prunellea possesses strong antioxidation with strengthen of SOD vigor and degradation of MDA;Establishment of anti-multiplication evaluation models:This topic studies the ability of Prunella vulgaris L.on the regulation of mitotic cycle and the mechanism of inducing apoptosis.Results showed that Prunella vulgaris L.could change the phase distribution on the cell cycle of lung cancer cell,including G0/G1 ratio increased from 34.30%to 54.59%,S phase decreased from 49.63%to 35.65%,and apoptosis ratio increased from 2.07%(125μg crude drug/mL) to 28.57%(1000μg crude drug/mL).Annexin V-FITC was also used to determine the cell apoptosis ratio,which show apoptosis ratio increasing from 17.1%(125μg crude drug/mL) to 44.5%(1000μg crude drug/mL).Establishment of the model for the modulation ability of immunology system:Take the TNF-αas the investigation target to evaluate the modulation ability of immunology system of Prunella vulgaris L..Results showed P.asiatica Nakai from Jiangsu and Prunella vulgaris L.from Anhui had the strongest ability,and the TNF-αcontent were 81.46±11.01,84.31±11.81 pg/mL,respectively.②In vitro screening system:In this topic,two kinds of human lung adenocarcinoma cell models:A549 and SPC-A-1 were established for the screening of spica prunellea.The consequence indicated that P.asiatica Nakai from Jiangsu and Prunella vulgaris L.from Anhui both had the strongest inhibition ability on the lung cancer cell A549 (IC50=62.98,65.33μg crude drug/mL) and SPC-A-1(IC50=113.54,116.47μg crude drug/mL).Experiments for the screening of dissolvant part indicated that XKC-60 had the strongest inhibition ability to lung cancer and then XKC-95,whose IC50 were 650.36μg crude drug/mL and 687.79μg crude drug/mL respectively.According to outcomes of dissolvant part’s compatibility,XKC-95 + XKC-60+ XKC-30+ XKC-W was the best one (IC50=514.65μg crude drug /mL).Based on the activity screening of every composition, phenolic acid +flavonoids was the optimal part,and then terpenoid,IC50 was 833.94,918.13μg crude drug/mL respectively.After composition of compatibility,it showed that terpene+ phenolic acid+flavonoids+polysaccharide+washing liquid position had the strongest activity(IC50=397.48μg crude drug/mL).All the results above manifested that the established cell models could be used to evaluate the activity of prevention and therapy for lung cancer.③The in vivo animal screening system:Building heterotransplantation lung cancer model,vaccinated Lewis cell on the C57 mouse,and chemical induction of A/J mouse lung cancer model.They were used to estimate the ability of spica prunellea on the prevention and therapy on lung cancer.Results indicated that P.asiatica Nakai from Jiangsu and Prunella vulgaris L.from Anhui had better inhibition ability on tumor-beating mice and the inhibition tumor rate were 60.05±6.76%,63.56±6.79%respectively.Terpenoid,phenolic acid, flavone and polysaccharide showed the best effection on the prevention and therapy of lung cancer. The inhibition tumor rate were 46.83±8.38%,44.91±5.73%and 45.04±5.39%, respectively.After compatibility of composition,terpenoid + phenolic acid + flavone show the stronger ability on the prevention and therapy of lung cancer,however,the inhibition tumor rate decreased after adding polysaccharide composition.The inhibition tumor rate were 55.27±5.34% and 50.93±3.11%,respectively.Results showed the established in vivo animal screening models could evaluate the prevention and therapy activity of lung cancer of spica prunellea..2.the material basis of spica prunellea’s prevention and therapy abilityof lung cancer:"constituent structure theory"Based on the research system of traditional Chinese medicine and the integrity,we proposed a hypothesis that the foundation of traditional Chinese medicine material is an ordered whole,has"constituent structure theory" with "three levels multi-dimensional structures".The dissertation has carried out the preliminary study of the material basis of spica prunellea on the prevention and treatment of lung cancer.Combined of HPLC,GC and DAD to analyze the composition compatibility of the material basis.Results showed fixed construction ratio in the material basis of spica prunellea.①Composition compatibility of material basis:prevention and therapy material basis of spica prunellea on lung cancer was composed with terpenoid,phenolic acid,flavone and polysaccharide based on the screening system.The composition compatibility possessed stable constituent ratio.The constituent ratio of flavone,phenolic acid and terpenoid of P.asiatica Nakai from Jiangsu and Prunella vulgaris L.from Anhui,which had the strongest medicine efficacy,were(1.41:0.44:0.53),(1.42:0.44:0.66),respectively.②Constituent ratio of main active ingredients in every compositions:primary explore the Constituent ratio of main active ingredients in every compositions.Results indicated that there was a stable constituent ratio among ursolic acid,oleanolic acid,caffeic acid,rosmarinci acid,rutin and quercetin.The constituent ratio of ursolic acid,oleanolic acid,caffeic acid,rosmarinci acid,rutin and quercetin of P.asiatica Nakai from Jiangsu and Prunella vulgaris L.from Anhui,were (1.0:14.7:3.9:1.0:4.4:1.4) and(1.0:14.8:4.0:0.8:5.6:1.8) respectively.