The Study Of MiR-222-3p Gene Silencing Participates In Pyroptosis Via NLRP3 Inflammatory Body-Caspase-1 And Intervention Of Jianpi Qingre Huoxue Decoction On Colon Carcinoma-bearing Mice

Aim: By observing the changes of NLRP-3,Caspase-1 and pyrolysis rate in different groups of tumor-bearing mice,explore the possible mechanism of Jianpi Qingre Huoxue Decoction to interfere with inflammation-related carcinogenesis.Method: Purchase 70 SPF-grade 4-6 weeks old BALB/c(nu)nude mice,randomly divided into 7 groups: silent group(positive control group),NC group,low-dose Chinese medicine group,middle-dose Chinese medicine group,high-dose group Chinese medicine group,chemotherapy group,and Chinese medicine chemotherapy combined group,each with 10 animals.The silent group was inoculated with the stable silencing miRNA-222 HCT-116 colon cancer cell line,and the other groups were inoculated with the HCT-116 colon cancer cell line in the armpit to prepare the colon cancer model of tumor-bearing mice.From the 2nd day of tumor transplantation,nude mice in the model group and the stable silent group were fed simultaneously,and the same dose of normal saline was administered continuously for 6 weeks;low,medium,and high doses were given Qingrejianpi prescription,and the chemotherapy group was given oxaliplatin Intraperitoneal injection.All animals were sacrificed at the end of the 6th week,and the tumor tissues under the armpits were taken for later use,and the tumor size was measured.Use microscope to observe HE stained tumor tissue and observe structural changes.Western-blot method and real-time fluorescent quantitative PCR method were used to detect the expression of NLRP3,Caspase-1 protein and mRNA in tumor tissue of colon cancer tumor-bearing mice.Application of TUNEL immunofluorescence double staining to detect the changes of tumor tissue pyrolysis.Reoults: The survival rate of each group was compared with 100% in the Chinese medicine group and the miR-222 group,80% in the chemotherapy group and the Chinese medicine chemotherapy combined group,and 60% in the model group.In terms of tumor volume,compared with the NC group(1.76±0.24),the tumor volume of the Chinese medicine group was reduced to varying degrees.The volume of the Chinese medicine low-dose group was1.53 ±0.1,and the Chinese medicine middle-dose group was 1.44±0.18 Chinese medicine high-dose group It was 1.34±0.25.The tumor suppression was the most obvious in the chemotherapy group and the Chinese medicine chemotherapy combined group.The chemotherapy group was 0.43±0.09,and the Chinese medicine chemotherapy group was 0.41±0.13.Compared with HE staining in each group,pathological mitosis was more common,and there was no significant difference.Compared with the NC group,the protein and gene expression of NLRP-3 and Caspase-1 in the traditional Chinese medicine group showed a downward trend.The difference was statistically significant(P<0.05).Compared with the NC group,the pyrolysis rate of each group was higher in the traditional Chinese medicine group.The difference was statistically significant(P<0.05).Conclusions : Jianpi Qingre Huoxue Recipe can inhibit the expression of miR-222,down-regulate the expression of NLRP-3 and Caspase-1,promote pyrolysis,increase survival rate,play the role of survival with tumor,and prevent inflammation-related cancer.