ABSTRACT
ObjectiveTo investigate the intervention effect of total glucosides of paeony （TGP） on the renal injury of MRL/lpr mice based on the Toll-like receptor 9 （TLR9）/myeloid differentiation factor 88 （MyD88）/nuclear transcription factor-κB （NF-κB） signaling pathway and explore the immunological mechanism of TGP in preventing and treating systemic lupus erythematosus （SLE）. MethodMRL/lpr female mice of SPF grade were randomly divided into a model group， a dexamethasone group （0.15 g·kg-1）， and high- （0.078 g·kg-1） and low-dose （0.039 g·kg-1） TGP groups， and female C57BL/6J mice were assigned to a blank group， with 7 mice in each group. Mice in each group were treated with corresponding drugs or normal saline by gavage at the same time every day. After 4 weeks， samples were collected. The kidney and spleen were weighed， and the organ index was calculated. Serum creatinine （SCr） and blood urea nitrogen （BUN） levels in each group were detected by biochemical assay. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes in the kidney. The degree of renal fibrosis was evaluated by Masson staining. The serum levels of interleukin （IL）-2， interferon （IFN）-α， IL-4， and anti-nuclear antibody （ANA） were detected by enzyme-linked immunosorbent assay （ELISA）. The mRNA expression of TLR9， MyD88， and NF-κB p65 in renal tissues was detected by real-time quantitative polymerase chain reaction （Real-time PCR）. The protein expression of TLR9 and NF-κB p65 in renal tissues was detected by immunofluorescence. The protein expression of TLR9， MyD88， and NF-κB p65 in renal and spleen tissues was tested by Western blot. ResultCompared with the blank group， the model group showed increased SCr， BUN， spleen index， and kidney index （P<0.05）， deteriorated pathological injury and fibrosis in renal tissues， elevated serum levels of IFN-α， IL-4， and ANA， decreased level of IL-2 （P<0.05）， and up-regulated TLR9， MyD88， and NF-κB p65 mRNA and protein levels in the kidney and spleen （P<0.05）. Compared with the model group， the TGP groups displayed reduced SCr， BUN， spleen index， and kidney index （P<0.05）， relieved pathological damage and fibrosis in renal tissues， decreased serum levels of IFN-α， IL-4， and ANA （P<0.05）， increased level of IL-2， and declining mRNA and protein expression levels of TLR9， MyD88， and NF-κB p65 in the kidney and spleen （P<0.05）. ConclusionTGP may inhibit the expression of downstream inflammatory factors to regulate immunity and resist SLE-induced renal injury by regulating the TLR9/MyD88/NF-κB signaling pathway.
ABSTRACT
Atherosclerosis （AS） is a chronic inflammatory disease which has great threat to human health in our society. The coronary heart disease， stroke and other cardiovascular and cerebrovascular diseases caused by it have a high mortality and disability rate. AS is characterized by endothelial dysfunction， lipoproteins accumulation and inflammatory cell infiltration， with a complex pathological mechanism during its evolution. Inflammatory response runs through the whole process of occurrence and development of AS， and is a common link of many pathological factors. Intervention of inflammatory response is an effective way and important strategy to control AS. By virtue of its unique advantages， traditional Chinese medicine （TCM） plays an important role in the clinical treatment. According to the TCM theory， the formation and development of AS are closely related to five factors， namely wind， fire， phlegm， blood stasis and deficiency. Among them， blood stasis and turbid phlegm are the most closely related. Blood stasis and turbid phlegm are both pathological products and pathogenic factors. Their formation is mainly due to the imbalance of Yin and Yang in the whole body， dysfunction of viscera and organs and obstruction of transfusion of Qi， blood and body fluid. The pathogenic factors are not only independent， but also related. In recent years， there are more and more studies on the intervention effect and mechanism of TCM on AS， but the intervention effect of TCM on the inflammatory response signaling pathways has not been reported. In this paper， it mainly introduces the understanding of the effect of TCM against AS and seven signaling pathways relating to the intervention effect of TCM on the inflammatory response， which are Janus kinase/signal transduction and activator of transcription （JAK/STAT） signaling pathway， nuclear factor-κB （NF-κB） signaling pathway， mitogen-activated protein kinase （MAPK） dependent signaling pathway， reactive oxygen species （ROS） dependent signaling pathway， CD40-CD40L signaling pathway and Toll-like receptor （TLR） dependent signaling pathway.
ABSTRACT
Objective: The effect of Xiexintang on Toll like receptor 9 (TLR9) signaling pathway in macrophage derived foam cells was studied by in vitro cell experiments. Method: The fifty SPF male SD rats were randomly divided into low, medium and high dose groups (1.4,4.2,12.6 g·kg-1·d -1) and normal groups. Except 20 rats in the normal group, 10 rats in each group were given equal volume of pure water gavage in the blank group. After the last Administration for 7 days, serum was separated,and the serum containing drugs in the low, medium and high dose groups of Xiexintang was prepared. Oxidized low density lipoprotein (ox-LDL) was used to intervene the differentiation of RAW264.7 macrophages into foam cells. The cell foam was identified by oil red O staining. After observing the effect of drug containing serum on the proliferation of macrophage derived foam cells by methye thiazolye telrazlium(MTT) method,the serum containing 20%concentration of each drug was selected to act on the foam cell model. The expression of interleukin(IL) -1β and interferon (INF) -γ was determined by enzyme-linked immunosorbent assay (ELISA) and real-time fluorescence quantitative PCR (Real-time PCR). TLR9, myeloid differentiation factor 88(MyD88)and nuclear factor(NF) -κB were detected by Western blot. Result: Oil red O staining showed that the red particles were obvious after ox-LDL intervention. The foam cell model was successfully prepared. MTT results showed that there was no significant difference in cell proliferation between the high dose group of Xiexintang in the 10%~30%concentration range and the normal group serum. Follow up selection of the serum containing 20%concentration of each dose intervened the foam cells induced by ox-LDL. Compared with the normal group,the model group after ox-LDL intervention induced the high expression of TLR9,MyD88,NF-κB p65,IL-1β,INF-γ (PPκB p65,IL-1β,INF-γ(PPConclusion: Xiexintang containing serum can inhibit ox-LDL-induced RAW264.7 macrophage foaming,and its mechanism may involve regulation of TLR9/MyD88/NF-κB p65 signaling pathway and inhibition of IL-1β and INF-γ overexpression. This may be one of its mechanisms of against atherosclerosis.
ABSTRACT
Diagnostic ions filter method was used to rapidly detect and identify the phenolic compounds in Rheum palmatum based on ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MSE). The representative authentic standards of phenolic compounds, including gallic acid, (+)-catechin, (－)-epicatechin, (－)-epicatechin-3-O-gallate and procyanidin B2, were subjected to analysis by UPLC-Q-TOF/MSE system with negative ion mode. Fragmentation patterns of each standard were summarized based on assigned fragment ions. The prominent product ions were selected as diagnostic ions. Subsequently, diagnostic ions filter was employed to rapidly recognize analogous skeletons. Combined with retention time, accurate mass, characteristic fragments and previous literature data, the structures of the filtered compounds were identified or tentatively characterized. A total 63 phenolic compounds (36 phenolic acid derivatives, 8 flavonoid derivatives and 19 tennis derivatives) in R. palmatum were identified, including 6 potential new compounds. The method of diagnostic ions filter could rapidly detect and identify phenolic compounds in R. palmatum This study provides a method for rapid detection of phenolic compounds in R. palmatum and is expected to complete the material basis of rhubarb.