| Ulcerative colitis related colon cancer(UCRCC)is one of the serious complications of ulcerative colitis(UC)and it is an important cause of death in UC patients.As the inflammation progresses,the risk of UC becoming cancerous gradually increases.Therefore,it is particularly important to intervene in people with higher risks.For UCRCC treatment,surgical resection and chemotherapy are often used clinically as the main treatment method.However,the currently used chemotherapeutic drugs have large adverse reactions,poor selectivity,and are susceptible to drug resistance.Traditional Chinese medicine is characterized by its multiple components and multiple targets.Advantages in the treatment of inflammation-related cancers.Gegen Qinlian Decoction showed good anti-UC effect in the early stage,which mainly through the NF-?B pathway to achieve intervention.The NF-?B pathway is also an important pathway for UCRCC.Therefore,the effect of Gegen Qinlian Decoction on the efficacy and main chemical components of UCRCC was studied in this paper in order to provide the basis for the follow-up of the spectrum-effect relationship,efficacy substances and quality control.(1)The mouse UCRCC model was established by AOM/DSS method,and the model was validated by hematoxylin-eosin staining.The results showed that the first cycle of the mice in the model group was over and the mucosal glands were destroyed extensively.The epithelial cells fell off and superficial ulcers formed;part of the glands were observed to be disordered and inflammatory cells infiltrated in the mucosal layer;at the end of the second cycle,the arrangement of the ducts was slightly disordered,the nucleus was deeply stained,and some of the nucleuses became elongated;the third cycle ended,we can see that the tumor ducts are arranged disorderly,there are many different sizes of cancer nodules on the surface of the colon.The colon cells are loosely arranged and the nuclei are dark blue.With the prolongation of the modeling period,precancerous lesions such as infiltration of inflammatory cells,disordered glandular arrangement,and atypical hyperplasia have occurred.The above results indicate that the UCRCC model has replicated successfully.(2)The preliminary study found that Gegen Qilian Decoction can improve UC,mainly through the NF-?B pathway to achieve intervention,NF-?B is also an important pathway for UCRCC,UCRCC's development was established on the basis of UC,it was speculated that Gegen Qilian Decoction control The NF-?B pathway affects UCRCC.Therefore,this part firstly uses the method of network pharmacology to establish an interactive network map of NF-?B and UCRCC through Cytoscape software,and selects NF-?Bp65,iNOS,COX-2 and Bcl-2,TNF-?,IL-1?,ICAM-1 and VCAM-1,according to Degree and Betweenness indicators.8 key inflammatory factors in the top overall ranking;using HE staining,immunohis-tochemistry,and enzyme-linked immunosorbent assay,Gegen Qilian Decoction on the above key inflammatory effect of factors.Hematoxylin/eosin(HE)staining was used to evaluate the degree of dysplasia of the mouse colonic epithelium.To explore the possible mechanisms of Pueraria Qilian Decoction in inhibiting colitis-associated colon cancer.The expression rates of NF-?Bp65 positive cells in the normal group,the model group,the high-,middle-,and low-dose groups of the Gegen Qilian Decoction and the 5-ASA intervention model group were 10.89±1.10,34.90±2.34,18.84±2.30,19.24±1.92,24.30±1.54 and 19.81±1.82%,respectively.The positive cell expression rates of iNOS were:9.40±1.80,37.80±5.31,17.60±3.10,18.40±6.32,24.40±4.50 and 18.31 ±2.00%,respectively.The positive cell expression rates of Bcl-2 were:11.50±1.30,46.80±5.93,20.32±2.80,22.80±2.72,27.30±3.64 and 21.86±2.48%,respectively.The positive cell expression rates of COX-2 were:8.40±1.10,26.30±6.83,1.9.40±2.30,19.70±1.64,21.20±1.32 and 18.60±2.700%,respectively;high-,middle-,and low-dose groups and model groups of Gegen Qinlian Decoction.Compared with the positive expression of NF-KBp65,iNOS,COX-2 and Bcl-2 in the colon tissue of mice,there was a significant difference in the expression of NF-?Bp65,iNOS,COX-2,Bcl-2 and the expression level of each drug intervention group was significantly lower than that of the model group(p<0.01).Enzyme-linked immunosorbent assay was used to detect serum TNF-? levels in the blank group,the model group,the high-,middle-,and low-dose group and the 5-ASA intervention model group mice 50.34±17.36,100.22±16.46,76.44±15.48,78.38±13.79,88.63±10.37 and 92.93±15.38ng/mL,respectively;IL-1? content were 28.16±17.45,31.81±29.29,20.41±11.06,30.25±12.98,42.30± 15.93 and 17.64±11.05ng/mL,respectively.The cont-ents of ICAM-1 were 47.48±9.19,108.04±16.75,63.98±16.98,86.99± 12.37,97.97±9.34 and 83.65±11.18ng/mL,respectively.The VCAM-1 contents were 5.35±4.28,19.78± 15.60,14.55±9.58,18.77±5.97,19.44± 12.60 and 19.19±6.51ng/mL,respectively.TNF-?,IL-1?,ICAM-1 and VCAM-1 in mice of Gegen Qinlian Decoction group were decreased compared with the model group.And there was a very significant difference compared with the model group(p<0.01).The above experiments have fully proved that Gegen Qinlian Decoction has a good intervention effect on UCRCC.(3)The fingerprint of Gegen Qilian Decoction was established by HPLC.On the basis of preliminary experiments,two elution systems were optimized,and the best elution system for the fingerprint of Gegen Qilian was determined to be acetonitrile 0.2%phosphoric acid.Water,gradient elution conditions:Omin:15%acetonitrile,10min:30%acetonitrile,25min:40%acetonitrile,40min:80%acetonitrile,50min:100%acetonitrile,55min:15%acetonitrile:optimum detection wavelength is 250nm.On this basis,the main chemical constituents of Pueraria sinensis were assayed,and the methodological investigation results showed that the linear relationship,precision,repeatability and stability were all good;puerarin,daidzin,glycyrrhizin,jatrorrhizine,baicalin,The recovery rates of palmatine,berberine,baicalein,glycyrrhizic acid and wogonin were 100.05±0.07,100.20±0.87,99.84±0.93,99.92±1.73,100.08±1.69,99.66±1.77,100.37±0.88,99.21±1.96,99.97±0.42 and 97.92±1.65%,respect ively;puerarin,daidzein,glycyrrhizin,jatrorrhizine,baicalin,palmatine,berberine,baicalein,glycyrrhizic acid,scutellarin.The contents of the hormones were 1.677±1.45,0.154±0.21,0.159±0.31,0.045±0.15,0.448±2.48,0.035±0.26,0.095±0.25,0.013±0.10,0.111 ±0.31 and 0.006±0.06?g/g,respectively.In summary,this experiment studied the pharmacodynamics of UCRCC and the chemical composition of Gegenqinlian decoction by Gegen Qinlian Decoction,which can lay the foundation and provide basis for future research on the relationship of spectrum effect,effective substances and quality control. |
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