Mechanism And Study Of YYFZBJS On Intestinal Adenoma

Colorectal adenoma is a benign intestinal tumor and an important precancerous le sion of colorectal cancer.More than 90%of intestinal cancer is developed from intestinal adenoma.Traditional Chinese medicine has a good clinical effect in the prevention and treatment of precancerous lesions of colorectal cancer.Therefore,this study examined the effects of the treatment of intestinal fistula on the occurrence and growth of colorectal adenomas,and the immunomodulation of APC gene mutations spontaneously in the formation of intestinal adenomas by observing the treatment of intestinal fistula in the treatment of intestinal sputum.Role;to explore whether its mechanism of action inhibits the occurrence and de velopment of colorectal adenoma by regulating T lymphocytes,and provides an e xperimental basis for the clinical application of YYFZBJS in the prevention and tr eatment of colorectal cancer.Objective:?1?C57BL/6J female mice were co-cultured with APC^Min/+male mice,genotype identification was performed by PCR nucleic acid electrophoresis experiments,and APC^Min/+mice were selected from the progeny mice for subsequent experiments.?2?The experimental mice were randomly divided into groups,and the rats were continuously intragastrically administrated with YYFZBJS for 20 weeks,and the body weight of the mice was weighed every week.After 20weeks,peripheral blood, spleen,intestine and mesenteric tissues of the mice were taken,and the number of intestinal adenomas was counted for statistical analysis.?3?The degree of carcinogenesis of intestinal adenoma was identified by HE staining; the expression of Ki67 and PCNA in intestinal tumor tissues was detected by immunohistochemistry,and the proliferation cells of intestinal adenoma were labeled with Brd U.?4?The transcription level of T lymphocyte-related genes in intestinal adenoma of APC^Min/+mice was analyzed by real-time PCR.?5?ELISA was used to detect immune-related cytokines in peripheral blood of APC^Min/+mice.Flow cytometry was used to sort and analyse the helper T lymphocytes in lymphocytes of mouse spleen,mesentery and small intestine lamina propria.?6?CCK-8 method was used to detect the effect of YYFZBJS on the proliferation of HCT116 and MC-38 cells,and the trypan blue test was used to detect the effects of YYFZBJS on lymphocytes in spleen and peripheral blood of mice.?7?The effects of sputum aconite and APC^Min/+mouse spleen Treg cell supernatant on the growth of MC-38 cells and the expression of related proteins were detected by trypan blue assay,colony formation assay and western-blot assay.Result:?1?Compared with the model group mice,the number of small intestine adenomas and the number of colorectal adenomas were significantly decreased after 20 weeks of administration of sputum aconite and sputum.?P<0.01?.The pathological results showed that the mice in the model group showed carcinoma in situ in the large intestine,and typical adenocarcinoma in the small intestine with local submucosal invasion;the mice in the aconite of the Fufuzi Sauce group showed no carcinoma in situ,and the small intestine was a high-grade gland.More common tumors,suggesting that YYFZBJS can effectively reduce the degree of malignant transformation of intestinal adenoma.IHC results showed that the positive expression rate of Ki67 and PCNA protein was significantly lower than that of the model group?P<0.01?.?2?Fluorescence quantitative PCR results of intestinal adenoma tissues of APC^Min/+mice showed that the m RNA expression levels of T-bet and ROR-?in the treatment group were higher than those in the model group,while the m RNA expression levels of Gata3 and Foxp3 were decreased?P<0.01?.Flow cytometry analysis of spleen,mesenteric lymphocytes and small intestine lamina propria of APC^Min/+mice showed that compared with the model group,the Treg cells in the treated group were significantly reduced;the levels of IL-10 and IL-6 in the serum of the mice were significant.The levels of IL-17A and TNF-?were significantly increased?P<0.01?,and showed a dose-dependent effect with the increase of the concentration of Aconite.It is suggested that the sputum aconite can enhance the cell function of Th1 and Th17,but inhibit the cell function of Th2 and Treg.?3?The results of CCK-8 in vitro showed that there was no significant inhibitory effect on the growth of HCT116 cells and MC-38 cells,and the difference was not statistically significant?P>0.05?.The trypan blue assay was used to detect the sputum Sauce powder has no inhibitory effect on the proliferation of peripheral blood and spleen lymphocytes in mice.?4?Trypan blue viable cell counting experiment and colony formation experiment showed that the supernatant of YYFZBJS can inhibit the proliferation of MC-38 cells after intervention of Treg cells;the results of Western-blot experiment showed that compared with the control group,The expression of Ki67 and PCNA protein in the drug intervention group decreased,and the difference was statistically significant?P<0.01?.Conclusion:?1?YYFZBJS can not improve the quality of life of APC^Min/+mice and reduce the number of intestinal adenomas.?2?YYFZBJS can reduce the positive expression rate of Ki67 and PCNA protein in intestinal tissues of APC^Min/+mice,and reduce the differentiation level of intestinal adenoma cells.?3?YYFZBJS can change the expression of lymphocyte-related factors in spleen,mesentery and intestine of APC^Min/+mice,enhance the cell function of Th1 and Th17,and inhibit the cell function of Th2 and Treg.The proportion of helper T lymphocytes in the spleen,mesentery and small intestine lamina propria lymphocytes of the treated group was reduced.Down-regulate the expression of IL-6,IL-10 and other tumor suppressor cytokines,and increase the expression of tumor suppressor cytokines such as TNF-?and IL-17A.?4?YYFZBJS can inhibit the growth of MC-38 cells and the expression of related proteins Ki67 and PCNA by regulating Treg cells.