Preliminary Study On The Effect Of Feifufang On Improving Cisplatin-induced Gastric Mucosal Injur

Objects:1.To study the effect of Feifufang on spleen and stomach function of Lewis lung cancer model mice with cisplatin.2.Calculate the key components and molecular mechanism of Feifufang in the treatment of gastritis and gastric ulcer.3.Verify the compatibility of key components of Feifufang in the treatment of gastritis and gastric ulcer on cisplatin induced injury of human gastric epithelial cells.Methods:1.Replicate the mouse model of Lewis lung cancer with deficiency of Qi and Yin:Ephedra,radix aconiti,Asarum and Rhizoma arisaematis decoction pieces were used to prepare thermal Chinese herbal decoction according to the mass ratio of 1∶1∶1∶1.Forty SPF C57BL/6J mice were randomly divided into blank group and model group according to body weight after adaptive feeding,including 8 blank group and32 model group,male and female in equal parts.Experimental animal model replication stage 1 mice were intragastally fed with hot Chinese herbal decoction combined with smoking for 14 days.On day 1 of the second stage of animal model replication,2 m L Lewis lung cancer cell suspension with a density of 2×10~7cells/m L was subcutaneously transplanted into the left forelimb under the armpit of the left forelimb of mice in the model group.After transplantation,the mice were routinely fed for 14 days.2.Effect of Feifufang on spleen and stomach function of Lewis lung cancer model mice with cisplatin:Thirty-two mice in the model group were randomly divided into model group,cisplatin group,Feifufang group and cisplatin+Feifufang group(combined group)according to gender and body weight after the replication of Lewis lung cancer model with Qi and Yin deficiency.Mice without modeling were set as blank group,with 8 mice in each group and half male and half female in 5 groups.Cisplatin group was intraperitoneally injected with 0.1 m L/10g on the 1st and 7th day of intervention.Feifufang group was given 0.3m L/10g by intragastric administration,once a day,for 14 days.The combined treatment group was given Feifufang(the same Feifufang group)and then cisplatin intraperitoneally(the same cisplatin group).The blank group was given 0.9%sodium chloride solution by intragastric administration at0.3m L/10g,once a day,for 14 consecutive days.The behavioral characteristics,water intake,food intake,body weight,tumor removal weight and organ index of Lewis lung cancer mice with qi and Yin deficiency were used as indexes.3.Using TCMSP,Swiss ADME,Uni Prot,Gene Cards,OMIM and Drug Bank,STRING and David databases and Cytoscape 3.7.2 constructs the network of"Feifufang-composition-gastritis/gastric ulcer"to predict the action mechanism of Feifufang in treating gastritis and gastric ulcer.PDB database and Auto Dock Tools were used for molecular docking of key components and key proteins.4.Study on the effects of the compatibility of key components of Feifufang in the intervention of gastritis and gastric ulcer on the damage of human gastric epithelial cells induced by cisplatin:Human gastric epithelial cells were cultured and divided into 7 groups,namely blank group,model group(16μmol/L cisplatin),positive drug group(0.4mg/m L aluminum magnesium carbonate),and 5,10,20,40μmol/L quercetin combined with 4μmol/L apigenin group,respectively.Cell survival rate,apoptosis fluorescence,mitochondrial membrane potential,ROS,GSH,CAT,SOD,MDA content and TNF-a,IL-1βexpression were used as indexes.Result:1.Lewis lung cancer cells were transplanted after 14 days of fumigation with hot Chinese herbal decoction,and then conventionally fed for 14 days,the tumor-bearing mouse model of Lewis lung cancer with deficiency of Qi and Yin was successfully created:The results showed that the mice in the model group showed mental fatigue,shortness of breath,black and purple tongue,and reduced water intake and food intake(P<0.05 or P<0.01).Transplantation of Lewis lung cancer cells showed fatigue,slow movement,dull hair color and increased body weight(P<0.05).2.Effect of Feifufang on spleen and stomach function of Lewis lung cancer model mice with cisplatin:Feifufang increased activity,water intake and body weight of Lewis lung cancer model mice with Qi and Yin deficiency(P<0.05 or P<0.01).Feifufang combined with cisplatin increased water intake,spleen and thymus index of Lewis lung cancer model mice with Qi and Yin deficiency(P<0.05 or P<0.01).3.According to the network pharmacology method,the components of Feifufang in treating gastritis include quercetin,luteolin and apigenin,etc.The components in treating gastric ulcer include quercetin,apigenin and maltoic acid,etc.15 key targets such as PTGS2,TNF-αand IL-1bwere obtained by calculating the interaction network between Feifufang and gastritis protein,and 101 key targets such as ACTB,AKT1 and IL-6 were obtained by calculating the interaction network between Feifufang and gastritis protein.Biofunctional enrichment analysis showed that there were 30 signaling pathways related to the treatment of gastritis by Feifufang therapy,including HIF-1,JAK-STAT,PI3K-AKT and NF-κB(P<0.0001).A total of 89signaling pathways related to lung compound therapy for gastric ulcer included PI3K-AKT,HIF-1,TNF and NF-κB(P<0.0001).The binding energies of quercetin and apigenin with TNF-aand IL-1bin the treatment of gastritis and gastric ulcer were all less than-5 Kcal/mol by molecular docking technique.4.Quercetin combined with apigenin inhibited cisplatin induced inflammatory response,oxidative stress and apoptosis of human gastric epithelial cells:compared with model group,quercetin combined with apigenin at different concentrations reduced ROS content(P<0.05 or P<0.01).20 and 40μmol/L quercetin combined with apenin increased the membrane potential and CAT content of JC-1(P<0.05).40μmol/L quercetin combined with apigenin increased the contents of GSH and SOD,and decreased the apoptosis fluorescence,MDA content and TNF-a,IL-1bexpression(P<0.05).