| Background:Inflammation is the body’s defense response to pathogens,harmful stimuli,or physical damage to tissues and cells.It is usually beneficial,limiting and removing harmful agents to repair damaged tissues.However,uncontrolled inflammatory responses can cause damage to the body,such as hypersensitivity diseases or inflammation in specific areas leading to lesions in parenchymal organs.In addition,long-term stimulation of chronic inflammation can also cause the occurrence and development of various diseases such as cardiovascular and cerebrovascular diseases,diabetes and tumors.Therefore,it is of great significance to carry out anti-inflammatory research.Traditional anti-inflammatory drugs are mainly western drugs,but they have side effects such as causing allergic reactions,strongly stimulating the digestive and respiratory systems,and damaging the liver and kidneys.However,traditional Chinese anti-inflammatory drugs have less drug toxicity and side effects than western drugs.Therefore,finding new TCM anti-inflammatory drugs and exploring their molecular mechanisms may lay a good foundation for the clinical application of drugs.TLR4,an important member of the Toll-like receptor family,can recognize and bind to the liposolysaccharide of Gram-negative bacteria and trigger inflammatory responses in the body.The inflammatory lesions of many organs and tissues are closely related to the expression level of TLR4 and its pathway-related proteins.Therefore,the study of TLR4 and its signaling pathway has become a hot topic in the field of anti-inflammation.Solanum lyratum Thunb(SLT)is a traditional Chinese medicine commonly and widely used in clinical practice which has anti-tumor,anti-allergy,antioxidant and many other pharmacological effects.It can be used in the treatment of a variety of diseases.There are abundant researches on the anti-tumor effects of SLT,but the anti-inflammatory studies on SLT are not systematic and in-depth at present.Therefore,based on the preliminary results of extraction and separation of SLT components,this study selected two extracts,SLT ferulic acid(sltPHA)and SLT saponin(sltSAP),to study their anti-inflammatory activities in vivo and in vitro and their effects on TLR4-targeted signaling pathway,so as to make full use of traditional Chinese medicine resources,effectively treat inflammatory diseases,and create economic and social benefits at the same time.Objective:To detect and evaluate the anti-inflammatory effects of sltPHA and sltSAP.To investigate the effect of sltPHA and sltSAP on TLR4-My D88-NF-κB signaling pathway.To identify the effective anti-inflammatory components of Solanum lyratum Thunb and develop candidate anti-inflammatory traditional Chinese medicine.Methods:CCK8 assay was used to investigate the effects of sltPHA and sltSAP at different concentrations on cell activity of mice macrophages(RAW264.7);The anti-inflammatory activities of sltPHA and sltSAP at cell level were studied and compared by ELISA kit and qPCR;The effects of sltPHA and sltSAP on acute abdominal inflammation induced by thioglycollate broth in mice were detected by blood smear cell count,white blood cell count and ELISA;QPCR,western blot and immunofluorescence were used to detect the effects of sltPHA and sltSAP on intracellular TLR4 expression;Western blot was used to detect the effects of sltPHA and sltSAP on the expression levels of proteins related to TLR4/My D88/NF-κB p65 signaling pathway.Results:1.Slt PHA had no significant effect on cell viability at 0.5,1 and 5μg/m L.Slt SAP had no significant effect on cell viability at 2,4 and 8μg/m L.2.Slt PHA and sltSAP treatment groups significantly reduced the secretion levels of inflammatory cytokines IL-1β,IL-6 and TNF-α in LPS-induced RAW264.7 cells(p<0.05)in a concentration-dependent manner.High concentration of sltPHA and sltSAP had the best anti-inflammatory effect,and sltPHA was better than sltSAP.3.Slt PHA and sltSAP at different concentrations significantly down-regulated the expression levels of IL-1β,IL-6 and TNF-α related factors in intracellular inflammation(p <0.05).High concentration of sltPHA and sltSAP had the best anti-inflammatory effect,and sltPHA was better than sltSAP.4.Different concentrations of sltPHA and sltSAP could significantly reduce the number of neutrophils in blood and white blood cells in abdominal fluid of mice with acute abdominal inflammation,and the difference was significant compared with model group(p <0.05),in a dose-dependent manner.High concentration of sltPHA and sltSAP had the best anti-inflammatory effect,and sltPHA was better than sltSAP.5.Different concentrations of sltPHA and sltSAP significantly reduced the expression of IL-1β and IL-6 in serum of mice with acute abdominal inflammation,and the difference was significant compared with model group(p <0.05),in a concentration-dependent manner.High concentration of sltPHA and sltSAP had the best anti-inflammatory effect,and sltPHA was better than sltSAP.6.High concentration of sltPHA and sltSAP could inhibit the expression of TLR4 protein and its downstream proteins My D88 and P65(p <0.05).Conclusion:1.Both sltPHA and sltSAP showed obvious anti-inflammatory effects,and the effectsof high concentration were the best.2.Both sltPHA and sltSAP exerted their anti-inflammatory effects by inhibiting TLR4-My D88-NF-κB signaling pathway.3.sltPHA has better anti-inflammatory effect than sltSAP. |
PDF Full Text Request