Study On The Regulatory Mechanism Of Baishaoqiwu Granules On TNBS-induced UC In NLRP3 Knockout Rats

Objective: To explore the therapeutic effect of BAISHAOQIWU granules on TNBS induced NLRP3 gene knockout rat UC model and its effect on related proteins and m RNA in colon tissue.Methods: 30 normal SD male rats and 30 NLRP3 knockout SD male rats,They were randomly divided into 6 groups by weight,Ten in each group,That is: A group is blank group: normal SD rats,B group was model group: normal SD rat s+TNBS,C group was BAISHAOQIWU granules group: normal SD rats +TNBS modeling + BAISHAOQIWU granules,D group was gene knockout blank group: NLRP3 gene knockout rats,E group was gene knockout model group: NLRP3 gene knockout rats +TNBS,F group was gene knockout medication administration group: NLRP3 gene knockout rats +TNBS modeling+BAISHAOQIWU granules,After making the model,The next day began to gavage.The rats in C、F group were perfused with water solution of BAISHAOQIWU granules containing 1.32 g/ml of raw drug,The other four groups were given equal volume saline,Each group was given once a day,The period of administration was 14 days.During the administration,All rats were given free drinking and feeding,Weigh once a day,The dosage was adjusted according to the change of body weight.Fasting and anesthesia on the last day.RATs were killed and specimens were collected for detection.Record animal weight,colon length,HE staining,pathology,PCR detection of colonic mucosa Caspase-1、IL-18、IL-1βm RNA expression,WB detect the expression of Caspase-1、IL-1β、IL-18 proteins in the colonic mucosa,To evaluate the effect of BAISHAOQIWU granules on NLRP3 gene knockout UC rats.Results: 1.DAI and CMDI scores: the scores of each group were significantly higher than those of blank control group and gene knockout blank group(P<0.05),BAISHAOQIWU granules group,gene knockout medication administration group,gene knockout model group scores were significantly lower than the model group(P<0.05),BAISHAOQIWU granules group and gene knockout medication administration group were significantly lower than gene knockout model group(P<0.05),BAISHAOQIWU granules group,gene knockout medication administration group score close(P>0.05).2.HE staining:inflammation was evident in the model group.Gland damage,Inflammation infiltrates the submucosa,There are a large number of fibrous connective tissue hyperplasia.The glands in the gene knockout model group were irregular,Vascular congestion and hemorrhage,Vascular dilatation and congestion in the lamina propriety and submucosa,a small amount of inflammatory cell infiltration.The pathological changes in the group of BAISHAOQIWU and the group of gene knockout medication administration were improved obviously,A slight amount of inflammatory cell infiltration in the mucosal layer,individual intestinal gland dilatation.3.Caspase-1、IL-18、IL-1βm RNA expression: the Caspase-1、IL-18、IL-1βm RNAexpression in the model group was significantly higher than that in the normal group(P<0.01);The expression of BAISHAOQIWU granules group,gene knockout medication administration group and gene knockout model group Caspase-1、IL-18、IL-1βm RNAsignificantly lower than that of model group(P<0.01);The expression of Caspase-1、IL-18、IL-1βm RNAin BAISHAOQIWU granules group and gene knockout medication administration group was significantly lower than that in gene knockout model group(P<0.01);Caspase-1、IL-18、IL-1βm RNAexpression was lower in gene knockout medication administration group than in BAISHAOQIWU granules group,but not statistically significant(P>0.05).4.The expression of Caspase-1、IL-18、IL-1β protein in colonic mucosa: model group,BAISHAOQIWU granules group,gene knockout medication administration group and gene knockout model group were higher than that in two normal group(P<0.05);Compared to the model group,The relative expression of Caspase-1、IL-18、IL-1β protein in BAISHAOQIWU granules group,gene knockout medication administration group and gene knockout model group were all low(P<0.05);Compared with the gene knockout model group,The relative expression of Caspase-1、IL-18、IL-1β protein in BAISHAOQIWU granules group and gene knockout medication administration group was low(P<0.05);However,there was no significant difference in the relative expression of Caspase-1、IL-18、IL-1β protein in the group of BAISHAOQIWU granules and gene knockout medication administration group(P>0.05).Conclusion: 1.The pathogenesis of rats UC induced by TNBS is related to the NLRP3 related signaling pathway,and knockout of NLRP3 genes will lead to the decrease of the severity of UC in rats.2.BAISHAOQIWU granules can inhibit the expression of Caspase-1、IL-18、IL-1β in colon tissue of UC rats by multiple targets,the main target of which is NLRP3 inflammatory signaling pathway.