Differential Analysis Of Fibrin Expression During The Occurrence Of Cold And Heat Colorectal Cancer

ObjectiveTumor microenvironment(TME)is a "complex network" that regulates the development of tumors.The remodeling of the extracellular matrix(ECM)plays an important role in tumorigenesis,development,invasion and metastasis.In order to investigate the correlation between the changes in the selection pattern of cold and heat syndrome colorectal cancer and the differences in fibrin expression,to observe the cold and heat syndrome colorectal cancer tissue fibrin family fibulin 4(Fibulin 4),fibulin 5(Fibulin 5)and the expression of lysyl oxidase(LOX),in order to find the similarities and differences of the tumor mechanism under different environmental pressures of cold and heat.MethodsWith reference to Xu Rufu and other groups with a completely random segment grouping method,156 Wistar rats(male and female)were divided into 12 blank groups,36 cold modules,and 36 cold-reverse syndrome group.There are 36 heat modules and 36 heat-reverse syndrome group.The cold model was administered with ice water at 0℃ and cold water bath at 10℃for cold modeling.The cold-reverse syndrome group was supplemented with Fan Zuo Jin Wan decoction on the basis of cold modeling.The heat model was modeled with 30%ethanol and capsaicin 0.9 mg·mL-1 solution;the heat-reverse syndrome group was added with Zuo Jin Wan decoction based on the heat model model;rats in the blank group were intragastrically given saline.At the 5th week of the experiment,32 rats(2 in the blank group and 6 in the remaining groups)were selected and tested for cold and heat modeling.At the 6th week,rats in the cold module,the cold-reverse syndrome group,the heat module,and the heat-reverse syndrome group were given subcutaneous injection of 25 mg · kg-1,once a week,for 12 consecutive times.Due to the anti-cancer effect of capsaicin,the heat module only gave 30%ethanol solution during the carcinogenesis,and the cold module maintained the model.All rats were given 10 mL·kg-1,once a day for 38 weeks.The materials were collected at 29,32,35,and 38 weeks to observe the incidence of cold and heat colorectal cancer,and Fibulin4,Fibulin5,and LOX in the blank group,cold module,cold-reverse syndrome group,heat module,and heat-reverse syndrome group expression differences in groups.Results1.The diet,drinking water,weight,and physical signs of rats with cold and heat modules are consistent with the symptoms of cold and heat syndromes described in "Practical Chinese Medicine Syndrome Animal Modeling".Symptom descriptions in the cold-reverse syndrome group and the heat-reverse syndrome group improved.The activity of cold module,heat module SDH,LDH,Na+K+-ATP and Ca2+Mg2+-ATP were significantly different.Compared with the cold and heat modules,the SDH,LDH,Na+K+-ATP,and Ca2+Mg2+-ATP of the cold-reverse syndrome group and the heat-reverse syndrome group were improved.The changes of gastric mucosa and intestinal mucosa were consistent with cold syndrome and heat syndrome.The gastric mucosa and intestinal tissue mucosa of cold-reverse syndrome group and heat-reverse syndrome group tended to improve.The results showed that the cold syndrome model and the heat syndrome model were established successfully,and the zuo jin wan and the fan zuo jin wan had positive effects on the cold syndrome and the heat syndrome.2.The cold and heat syndrome colorectal cancer models were established.By observing the pathological sections,it was found that the incidence of tumors in the cold-reverse syndrome and heat-reverse syndrome groups was lower than that of the cold modules and heat modules in the early colorectal cancer.The incidence of tumors in cold module was lower than that in heat module.When colorectal cancer develops to the middle stage,the occurrence of cold module tumors progresses slowly,and the occurrence of heat module tumors progresses rapidly.The tumor progression in the cold-reverse syndrome group and the heat-reverse syndrome group gradually accelerated.At the later stage,there was no significant difference in the incidence of tumors between the cold module and the cold-reverse syndrome group,and the heat module and the heat-reverse syndrome group.However,the occurrence and pathology of cold module tumors were more serious than that of heat modules.3.Immunohistochemical and Western Blot tests were performed on colorectal cancer tissues,and the results showed that the expression differences of Fibulin4,Fibulin5,and LOX in the cold module and cold-reverse syndrome groups,the heat module and heat-reverse syndrome groups were significantly different in the early stage of the tumor.In the late stage of tumor,the expression differences of cold module,cold-reverse group,heat module and heat-reverse group decreased.4.Fan Zuo Jin Wan and Zuo Jin Wan have positive effects on cold syndrome and heat syndrome,and can inhibit the occurrence of colorectal cancer in the early stage of colorectal cancer.This has a certain relationship with the medicinal properties and the ratio of the two prescriptions.Conclusion1.The cold syndrome model can be established by ice-water gavage and ice-water bath,and the hot syndrome model can be established by capsaicin and ethanol mixed solution.and the zuo jin wan and the fan zuo jin wan had positive effects on the cold syndrome and the heat syndrome.2.It is feasible to construct a model of colorectal cancer with cold syndrome and heat syndrome.After combining the disease and syndrome,the corresponding syndromes can be explored to explore the mechanism of colorectal cancer selection under different environmental pressures,providing a reliable basis for the study of TCM syndromes and the mechanism of tumor microenvironment Animal model experience.3.The stiffness of large intestine and the occurrence of tumor are related to the different internal environment of cold syndrome and heat syndrome.The intestinal tissues of the cold module showed higher hardness and brittleness,increased fibrosis,and obvious characteristics of promoting fiber proliferation.The hardness and brittleness of intestinal tissues appeared in the thermal module.4.Fan Zuo jin wan and zuo jin wan have contradictory effects on cold syndrome and heat syndrome,but their contralateral evidence weakens in later stages.The effect of pre-tumor treatment is better,and its effect is closely related to the drug ratio of zuo jin wan and fan zuo jin wan.However,there are still some risks in taking Chinese medicine for a long time,and the efficacy of Chinese medicine in the later stage is very small.