Study On Inhibitory Effects And Mechanism Of Yiai Capsule On Human Lung Cancer Cell NCI-H460 In Nude Mice

Objective:To study the effect of Yiai capsule on tumor growth inhibition of human lung cancer NCI-H460 cell xenograft in nude mice and its mechanisms,and to provide experimental evidence for the development and application of Yiai capsule in anti-tumor.Methods:1.40 Kunming mice were randomly divided into two groups:the administration group?n=20?:with 24 g/kg dose by intragastrical;the control group?n=20?:the same volume of distilled water by intragastrical.The mice were given 3 times within 24 hours by the maximum tolerance dose,all the mice were observed 14 days and record the weight every 7 days,then observed the acute toxicity reaction of Yiai capsule.2.12 SD rats were randomly divided into three groups:the blank group?n=4?:with equal volume of distilled water by intragastrical;the Yiai capsule group?n=4?:with 1.000 g/kg dose by intragastrical;the CTX group?n=4?:with 0.025 g/kg dose by intraperitoneal injection.To draw blood from abdominal aorta in one hour after the last intragastrical,separation of serum,the inactivated under 56?for 30min and stored at-20?.The inhibitory effects of serum containing Yiai capsule on NCI-H460,SMMC-7721 and HepG₂cells were detected by MTT assay in vitro.3.NCI-H460 cells were injected subcutaneously into nude mice to establish model of transplanted tumor.40 nude mice were randomly divided into five groups:the blank group?n=8?:with equal volume of distilled water by intragastrical;the high dose group of Yiai capsule?n=8?:with 3.600 g/kg dose by intragastrical;the middle dose group of Yiai capsule?n=8?:with 1.800 g/kg dose by intragastrical;the low dose group of Yiai capsule?n=8?:with 0.900 g/kg dose by intragastrical;the CTX group?n=8?:with 0.025 g/kg dose by intraperitoneal injection.The CTX group was administered every other day,the other groups were administered once a day.After 14 days,the nude mice were put to death,take the eyeball for blood and the tumor was ready to use.4.The number of RBC,hemoglobin count,PLT,WBC were counted by automatic blood analyzer.The contents of ALB,ALP,ALT,AST,BUN,CREA,GLB,TBIL and TP were counted by extract serum.IL-6 and TNF-?levels in serum were measured by ELISA.5.The content of SOD was determined by hydroxylamine method,and the content of MDA was determined by thiobarbituric acid method.6.The pathological changes of tumor tissue by HE staining were observed with light microscope.7.TUNEL method was applied to detect the degree of apoptosis tumor tissue after Yiai capsule treatment.Results:1.Acute toxicity test results show that compared with the control group,the weight of the dose group was not significantly reduced?p>0.05?.No obvious symptoms of poisoning and the mice all survived.2.MTT assay revealed that the Yiai capsule could inhibit the proliferation of NCI-H460,Hep G₂ and SMMC-7721 cells.The inhibitory effect on the NCI-H460 cells more significant than on Hep G₂,SMMC-7721 cells.In a certain time range,the inhibitory effect of serum containing Yiai capsule on cancer cells increased with the increasing of the time.3.The nude mice were all observed tumor appearancing.The inhibitory rates of the high dose group of Yiai capsule,the middle dose group of Yiai capsule and the low dose group of Yiai capsule were 36.3%,33.7%,18.0%respectively.Tumor weight in the high dose group of Yiai capsule and the middle dose group of Yiai capsule were significantly lower than that in blank group?P<0.05?.And no significant difference was found between the low dose group of Yiai capsule and the blank group in the tumor weight?P>0.05?.4.There were no significant difference in the number of WBC,hemoglobin and PLT among the high dose group of Yiai capsule,the middle dose group of Yiai capsule,the low dose group of Yiai capsule and the blank group?P>0.05?.The number of RBC in the high dose group of Yiai capsule and the middle dose group of Yiai capsule were lower than that in blank group?P<0.05?.5.There were no significant difference was found among the high dose group of Yiai capsule,the middle dose group of Yiai capsule and the low dose group of Yiai capsule and the blank group in the contents of ALB,ALP,ALT,AST,BUN,CREA,GLB,TBIL and TP?P>0.05?.6.The content of TNF-?in the high dose group of Yiai capsule,the middle dose group of Yiai capsule and the low dose group of Yiai capsule were significantly higher than that in the blank group?P<0.01?.The content of IL-6in the high dose group of Yiai capsule were significantly lower than that in the blank group?P<0.05?.7.The content of MDA in the high dose group of Yiai capsule,the middle dose group of Yiai capsule and the low dose group of Yiai capsule were significantly lower than that in the blank group?P<0.01?.The activity of SOD in the high dose group of Yiai capsule were significantly higher than that in the blank group?P<0.01?.8.The HE staining results showed that the cancer cells of the blank group arranged closely.Acellular necrosis of tumor tissue was more serious in the high dose group of Yiai capsule,the middle dose group of Yiai capsule and the low dose group of Yiai capsule than that in the blank group.9.The TUNEL staining results showed that apoptotic cells in the Yiai capsule group were more than those in the blank group.The apoptosis index of the high dose group of Yiai capsule,the middle dose group of Yiai capsule and the low dose group of Yiai capsule were 22.2%,16.2%and 11.0%respectively,there was a significant difference between the Yiai capsule group and the blank group?P<0.01?.Conclusion:The results suggest that Yiai capsule shows inhibitory effects on the growth of human lung cancer cell NCI-H460 in nude mice,which may be related to increased antioxidative ability and TNF-?content,as well as inhibition of IL-6 secretion.