健脾散积方通过LPS介导TLR4/MAPK通路抑制肠腺瘤生长的机制OA

Objective It is to observe the effect of Jianpi Sanji Decoction on colorectal adenoma in mice,and to explore the molecular mechanism in inhibiting the growth of colorectal adenoma in mice based on lipopolysaccharide(LPS)in in-flammatory microenvironment and its mediated Toll-like receptor 4(TLR4)/mitogen-activated protein kinase(MAPK)sig-naling pathway.Methods Thirty-two male C57BL/6 mice were randomly divided into blank group,model group,LPS group and Jianpi Sanji Decoction group,with 8 mice in each group.The mice of all groups except for the blank group were prepared three-cycle chemical induction with azoxymethane(AOM)and dextran sulfate sodium(DSS)to establish model of colorectal adenoma.From the first day to the end of the experiment,the Jianpi Sanji Decoction group was given Jianpi Sanji Decoction 27.53 g/kg(0.2 mL/mouse)by gavage,and the blank and model groups were intragastrically adminis-tered with the same volume of saline,all once daily;the LPS group was intragastrically administered with the same amount of saline and intraperitoneally injected with 0.1 mL LPS at the end of modeling,continuously treated for 3 days.The daily activities,body weight,stool characteristics and other general conditions of the mice were observed;the number of colorec-tal adenoma was recorded;the pathological morphology of colorectal tissues was observed by HE staining;the serum levels of LPS in the blank group,model group and Jianpi Sanji Decoction group were measured by ELISA;the mRNA expressions of RAS,BRAF and MAPK in colorectal tissues were detected by RT-qPCR;the protein expressions of TLR4,RAS,BRAF,extracellular signal-regulated kinase(ERK)and p-ERK in colorectal tissues were detected by Western blot;the positive protein expressions of RAS,BRAF and ERK in colorectal tissues were detected by immunohistochemistry.Results Compared with the control group,the mice in the model group all had symptoms such as weight loss,diarrhea with hema-tochezia,and listless,and with intestinal polyps of varying sizes;HE staining revealed that the colorectal polyps were high-grade intraepithelial neoplasia;the serum level of LPS was significantly increased(P<0.05);the relative mRNA expres-sions of RAS,BRAF and MAPK,relative protein expressions of TLR4,RAS,BRAF,ERK and p-ERK,and the mean op-tical density values of positive protein expressions of RAS,BRAF and ERK in colorectal tissue were all significantly in-creased(all P<0.05).Compared with the control and LPS groups,the physical condition of mice in the Jianspi Sanji De-coction group was improved,and the number of adenomas was significantly reduced(all P<0.05);the serum level of LPS(excluding the LPS group)was significantly reduced(P<0.05);the relative mRNA expressions of RAS,BRAF and MAPK,as well as the relative protein expressions of RAS,BRAF and p-ERK,and the mean optical density values of posi-tive expressions of RAS,BRAF and ERK in colorectal tissue were all significantly reduced(all P<0.05),while the rela-tive protein expression of TLR4 was significantly increased(P<0.05).Conclusion Jianspi Sanji Decoction can reduce the level of LPS in the inflammatory microenvironment and the activation of the downstream TLR4/MAPK signaling pathway mediated by LPS,thereby inhibiting the growth of colorectal adenoma in mice.