基于NLRP3/Caspase-1/GSDMD信号通路探讨首荟通便胶囊对便秘小鼠结肠组织损伤的影响OA

AIM To investigate the effects of Shouhui Tongbian Capsules on colon tissue injury in constipation mice.METHODS C57BL/6J mice were randomly divided into control group,model group,mosapride group(3 mg/kg mosapride citrate dispersible tablets)and Shouhui low-dose and high-dose groups(150,300 mg/kg Shouhui Tongbian Capsules),with 12 mice in each group.Except for the control group,mice in other groups were given 10 mg/kg loperamide hydrochloride by gavage for 21 days to induce constipation models.After 7 days of modeling,they were given drugs by gavage continuously for 14 days.The excretion time of the first black stool in mice was observed,and the fecal moisture content and intestinal propulsion rate were measured.HE staining was used to observe the pathological morphology of colon tissue.Serum levels of IL-1β,IL-18 and TNF-α were detected by ELISA.Immunohistochemistry was used to detect the protein expressions of IL-18 and NLRP3,and the intestinal barrier protein expressions of Claudin-1 and Occludin in colon tissue.Western blot was used to detect the protein expressions of p38,p-p38,NF-κB,IL-1β,IL-6,IL-18,TNF-α,cleved-Caspase-1(C-Caspase-1)+Caspase-1,GSDMD+N-GSDMD,NLRP3 and ASC in colon.RT-qPCR was used to detect the mRNA expressions of p38,NF-κB,IL-1β,TNF-α,Caspase-1,GSDMD,NLRP3 and ASC in colon tissue.RESULTS Compared with the control group,the excretion time of the first black stool in the model group was prolonged(P＜0.05),and the fecal moisture content and intestinal propulsion rate were decreased(P＜0.05).Colon tissue showed inflammatory injury and barrier damage.Serum levels of IL-1β,IL-18 and TNF-α increased(P＜0.05).The protein expressions of p-p38/p38,NF-κB,IL-1β,IL-6,IL-18,TNF-α,C-Caspase-1/Caspase-1,N-GSDMD/GSDMD,NLRP3 and ASC in colon tissue were increased(P＜0.05),while the protein expressions of Claudin-1 and Occludin decreased(P＜0.05).The mRNA expressions of p38,NF-κB,IL-1β,TNF-α,Caspase-1,GSDMD,NLRP3 and ASC in colon tissue increased(P＜0.05).Compared with the model group,the excretion time of the first black stool in Shouhui high-dose group was shortened(P＜0.05),and the fecal moisture content and intestinal propulsion rate were increased(P＜0.05).Inflammatory injury and barrier damage of colon tissue recovered.Serum levels of IL-1β,IL-18 and TNF-α decreased(P＜0.05).The protein expressions of p-p38/p38,NF-κB,IL-1β,IL-6,IL-18,TNF-α,C-Caspase-1/Caspase-1,N-GSDMD/GSDMD,NLRP3 and ASC in colon tissue decreased(P＜0.05),while the protein expressions of Claudin-1 and Occludin increased(P＜0.05).The mRNA expressions of p38,NF-κB,IL-1β,TNF-α,Caspase-1,GSDMD,NLRP3 and ASC in colon tissue decreased(P＜0.05).CONCLUSION Shouhui Tongbian Capsules can improve the inflammatory injury and barrier damage of colon tissue in constipation mice,and its effect may be related to anti-inflammation and inhibition of activation of NLRP3/Caspase-1/GSDMD signaling pathway.