| [Objective] In this study,the energy metabolic reprogramming in the evolution of IBD-CRC was used as the entry point,and the mouse model of IBD-CRC induced by AOM/DSS was selected as the research object.On the basis of verifying the effect of Buzhong Yiqi Decoction on the progression of IBD-CRC,the effects of energy metabolic reprogramming and key enzymes on metabolism were observed,and its molecular biological mechanism was further revealed.[Methods] 1.Established the mouse model of IBD-CRC induced by AOM/DSS,use AOM intraperitoneal injection(10mg/Kg)+ 2 cycles of DSS(3.0%)to make a stable,high repetition rate IBD-CRC mouse model,Observe the general condition of the mental state,reaction rate,hair luster and shedding,weight change,rectal prolapse,stool,etc.,and perform DAI score.On the last day of the 4th,5th,6th,and 7th week,the tissue was taken and the histomorphological changes were observed by pathological section H&E staining to evaluate the success of modeling.2.Effect of Buzhong Yiqi Decoction on preventing the evolution of IBD-CRC and its effect on the energy metabolic reprogramming: The mice were randomly divided into 6 groups: blank control group,model control group,positive control group,Buzhong Yiqi Decoction low-dose group,Buzhong Yiqi Decoction medium-dose group and Buzhong Yiqi Decoction high-dose group.40 mice in each group,each group began to be administered one week after modeling,and on the last day of the 4th,5th,6th,and 7th week,mice were randomly selected for picking up the eye to get blood and get colon tissue:(1)Observe the general condition of the mice,perform DAI score,and observe the pathological changes of colon tissue by H&E staining;(2)The levels of glucose and lactate(ELISA)and HK2,LDHA(Western blot)in colon tissue were measured to assess the energy metabolic reprogramming,and screened the dominant dose of Buzhong Yiqi Decoction.3.Energy-regulating mechanism of Buzhong Yiqi Decoction in blocking the progression of IBD-CRC: The mice were randomly divided into 4 groups: blank control group,model control group,positive control group and Buzhong Yiqi Decoction group(dominant dose).40 mice in each group,each group began to be administered one week after modeling,and on the last day of the 4th,5th,6th,and 7th week,mice were randomly selected for picking up the eye to get blood and get colon tissue:(1)Detection the expression of serum inflammation-related factor IL-6(ELISA);(2)Detection the expression of colonic nuclear factor STAT3(immunohistochemical staining);(3)Detection of the content of c-Myc protein in the metabolism of the colon(Western blot).[Results] 1.Successfully established the mouse model of IBD-CRC: After the mice began to drink DSS,they gradually showed weight loss,hair dryness,decreased activity,perianal blood,weight loss,and increased DAI score.Histopathological H&E staining showed: extensive inflammatory cell infiltration in the colon epithelium and lamina propria at the 4th week,suggesting that inflammation formed,the more the inflammation was closer to the anus;the fifth week,the crypt structure was disordered,the gland was deformed,the arrangement was disordered,and some were visible.Distorted crypt foci;at week 6,most of the mice showed dysplasia in the colon,large nuclear staining,large nuclear ratio,disordered cell arrangement,and extreme disappearance.At the 7th week,colonic tumor formation and tumor formation rate was 100%,suggesting the model established successfully.2.Observed the effects of Buzhong Yiqi Decoction on the general condition,body weight change,DAI,colonic lactic acid and glucose levels and key metabolic enzymes such as HK2 and LDHA in model mice of IBD-CRC,and evaluated the effect of Buzhong Yiqi Decoction on preventing the evolution of IBD-CRC and its effect on the energy metabolic reprogramming:(1)The positive group and Buzhong Yiqi Decoction in the high and middle dose groups were generally better than the model group.The mental state of the mice,the activity,the stool formation,and occasional blood in the stool,the weight loss was small.(2)In addition to the blank group,the DAI of the other groups increased,and the increase of DAI in the middle and high dose groups of the positive group and Buzhong Yiqi Decoction was small.(3)H&E pathological staining showed that the model group showed “inflammation-distortion crypt foci-dysplasiacancer” in the 4th to 7th week,and no obvious cancer formation was found in the Buzhong Yiqi Decoction group at the 7th week.(4)The positive group and Buzhong Yiqi Decoction medium and high dose group can significantly reduce the accumulation of lactic acid in the colon of mice and slow down the consumption of glucose,and can reduce the expression of HK2 and LDHA protein.It is suggested that Buzhong Yiqi Decoction can regulate energy metabolic reprogramming,inhibit aerobic glycolysis,improve the survival state of mice with colitis-induced transformation,reduce intestinal mucosal damage,and thus delay the progression of IBD-CRC,and middle dose is the dominant dose.3.The results of detecting serum IL-6 in mice and STAT3 and c-Myc in colon tissue showed:(1)Compared with the blank group,the serum IL-6 level in the model group was significantly higher(P<0.01).Compared with the model group,the IL-6 content in the positive group and Buzhong Yiqi Decoction group decreased,and the difference was statistically significant(P<0.01).(2)Compared with the blank group,the expression of STAT3 in the colon tissue of the model group was up-regulated.Compared with the model group,the expression of STAT3 in the colon tissue of the positive drug group and Buzhong Yiqi Decoction group was significantly down-regulated(P<0.01).(3)Compared with the blank group,the expression of c-Myc protein in the colon tissue of the model group was significantly increased,and the difference was extremely significant(P<0.01).Compared with the model group,the expression of c-Myc protein in the colon tissue of the positive group and the Buzhong Yiqi Decoction group was significantly reduced,and the difference was significant(P < 0.01).[Conclusion] 1.Buzhong Yiqi Decoction can effectively delay the progression of IBD-CRC,and the best effect is the medium.2.The benign effect of Buzhong Yiqi Decoction on IBD-CRC is through regulation of energy metabolic reprogramming.3.Buzhong Yiqi Decoction inhibits IL-6/STAT3/c-Myc signal transduction pathway,down-regulates the level of key metabolic enzymes HK2 and LDHA,thereby regulating energy metabolic reprogramming and intervening in IBD-CRC. |
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