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ObjectiveTo observe the effect of shikonin （SKN） on synovitis in DBA/1 mice with collagen-induced arthritis （CIA）. MethodThirty-six DBA/1 mice were randomly divided into a normal group， a CIA group， low-， medium-， and high-dose SKN groups （1， 2， and 4 mg·kg-1）， and a methotrexate （MTX， 0.5 mg·kg-1） group， with 6 mice in each group. Mice in the CIA group， the SKN groups， and the MTX group were immunized with an equal volume of bovine type Ⅱ collagen and complete Freund's adjuvant on day 1. On day 21， those mice received a second immunization with an equal volume of bovine type Ⅱ collagen and incomplete Freund's adjuvant to establish the CIA model. On the day of the second immunization， mice were treated with drugs by gavage. Mice in the MTX group received oral administration three times a week， while others received once per day， for 28 days. On day 22， the symptoms of arthritis， such as redness and swelling of joints， in CIA mice were observed， and arthritis scores were recorded. On day 49 after sample collation， histopathological examination of synovial inflammation in CIA mice was performed using hematoxylin-eosin （HE） staining. Immunofluorescence （IF） double labeling was used to detect the expression of vimentin and mitogen-activated protein kinase 1 （MAPK1） in the synovium of CIA mice. Network pharmacology predicted that the target of SKN in rheumatoid arthritis （RA） was MAPK1， which was verified by molecular docking. Western blot was used to detect the expression of extracellular signal-regulated kinase （ERK）， c-Jun N-terminal kinase （JNK）， p38， phosphorylated （p）-ERK， p-JNK， and p-p38 proteins in the synovial membrane of mice. ResultCompared with the normal group， the CIA group showed significantly higher arthritis scores， morbidity， and synovial inflammation， severely disrupted joint structure， evident articular cartilage and bone destruction， severe bone erosion （P<0.01）， increased expression of vimentin and MAPK1 in the synovium of mice， and increased protein expression of p-ERK/ERK， p-JNK/JNK， and p-p38/p38 in the synovium of mice （P<0.01）. Compared with the CIA group， the SKN groups and the MTX group showed relatively normal joint structure， with milder bone erosion and bone destruction， and smoother articular surfaces. Molecular docking results confirmed that the target of SKN was MAPK1. In the SKN groups and the MTX group， the expression of vimentin and MAPK1 in the synovial membrane was significantly reduced （P<0.01）， and the protein expression of p-ERK/ERK， p-JNK/JNK， and p-p38/p38 in the synovium of mice was significantly reduced （P<0.01）. ConclusionSKN can target MAPK1 to inhibit the protein expression of p-ERK， p-JNK， and p-p38 in CIA mice， thereby treating RA.
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ObjectiveTo observe the effect of Flemiphilippinin D on collagen-induced arthritis （CIA） in rats and explore its mechanism. MethodForty rats were randomly divided into normal group， CIA group， methotrexate （MTX） group （1.35 mg·kg-1）， low-dose Flemiphilippinin D group （1.5 mg·kg-1）， and high-dose Flemiphilippinin D group （3.0 mg·kg-1）， with eight rats in each group. Except for the normal group， the CIA model was induced by type Ⅱ collagen. Each group was given corresponding liquid medicine or normal saline， once a week in the MTX group， and once a day in the Flemiphilippinin D groups for a total of 28 days. The arthritis score and joint swelling degree of rats were experimentally recorded. Pathological changes in the ankle joint of rats were observed by hematoxylin-eosin （HE） staining. Serum levels of inflammatory cytokines interleukin （IL）-1β， IL-6， IL-8， and tumor necrosis factor （TNF）-α were detected by enzyme-linked immunoabsorbent assay （ELISA）， and the mRNA expression of Toll-like receptor 2 （TLR2）， myeloid differentiation factor 88 （MyD88）， and nuclear transcription factor-κB （NF-κB） p65 were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， and the protein expressions of TLR2， MyD88， and NF-κB p65 were detected by Western blot. ResultCompared with the normal group， the ankle joint of the CIA group was significantly swollen， and the clinical score of arthritis and the degree of joint swelling were significantly increased （P<0.01）. The ankle joint tissue structure was significantly damaged， and the levels of inflammatory factors IL-1β， IL-6， IL-8， and TNF-α in serum were significantly increased （P<0.01）. The mRNA levels and protein levels of TLR2， MyD88， and NF-κB p65 were significantly increased（P<0.01）. Compared with the CIA group， arthritis clinical score and joint swelling of rats in each administration group were significantly reduced （P<0.05， P<0.01）， and the pathological changes in the ankle joint were significantly improved. The contents of serum IL-1β， IL-6， IL-8， and TNF-α were significantly decreased （P<0.05， P<0.01）. The mRNA levels and protein levels of TLR2， MyD88， and NF-κB p65 in the ankle joint were significantly decreased （P<0.05， P<0.01）. ConclusionTo a certain extent， Flemiphilippinin D can reduce the expression of inflammatory factors in rheumatoid arthritis rats and play a good therapeutic effect. It works perhaps by inhibiting the activation of the TLR2/MyD88/NF-κB signaling pathway and thus shows an anti-inflammatory effect.
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ObjectiveTo investigate the protective effect of Jianpi Huogu prescription （JPHGP） on the functional injury of vascular endothelial cells caused by alcohol and explore its mechanism based on protein kinase B/c-Jun amino-terminal kinase/p38 MAPK （Akt/JNK/p38 MAPK） signaling pathway. MethodThrough chick embryo allantoic membrane， thoracic aortic ring， and migration， invasion， adhesion， and lumen formation of human umbilical vein endothelial cells （HUVEC）， the effect of JPHGP with different concentrations （8， 16 and 32 μg·L-1） on angiogenesis was observed in the presence or absence of alcohol. The expression levels of phosphorylation of Akt， JNK， and p38 MAPK were determined by Western blot. ResultAs compared with the normal group， the number and length of capillaries around the arterial ring in the model group were decreased， and the migration， invasion， and lumen formation capacity of HUVEC were decreased （P<0.05， P<0.01）. After treatment with 16 and 32 μg·L-1 JPHGP， the length of neovascularization in chick embryo allantoic membrane was significantly increased （P<0.05， P<0.01）. Compared with the model group， the 8， 16， and 32 μg·L-1 JPHGP groups increased the number of capillaries around the thoracic aortic ring in a concentration-dependent manner （P<0.05， P<0.01）， and the 32 μg·L-1 JPHGP group increased the length of capillaries around the thoracic aortic ring （P<0.05）. The 16 and 32 μg·L-1 JPHGP groups enhanced the migration， invasion， and lumen formation capacity of HUVEC. The results of Western blot showed that， as compared with the normal group， the protein expression levels of p-JNK/JNK， p-p38 MAPK/p38 MAPK， and p-Akt/Akt were significantly decreased in the model group （P<0.01）， and as compared with the model group， the protein expression levels of p-p38 MAPK/p38 MAPK and p-Akt/Akt were significantly increased in the 8， 16， and 32 μg·L-1 JPHGP groups （P<0.01） and the protein expression level of p-JNK/JNK was increased significantly in the 16 and 32 μg·L-1 JPHGP groups （P<0.01）. ConclusionJPHGP has a protective effect on the functional injury of vascular endothelial cells caused by alcohol， and its mechanism may be related to the activation of Akt/JNK/p38 MAPK signaling pathway. Relevant research results will provide certain scientific basis for clarifying the effect of JPHGP on 'invigorating spleen and promoting blood circulation'.
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Objective To study the influence of the combined administration of Bletilla striata and Radix aconiti preparata with different doses or ratios on anhydrous ethanol-induced gastric mucosal injury in mice with gastric ulcer. Methods The uniform design method was adopted on the basis of two factors and seven levels, which aims to investigate the influence of combined oral administration of Bletilla striata and Radix aconiti preparata with different doses or ratios on gastric mucosal injury in the mice with gastric ulcer. The indices like gastric ulcer index and stomach mucosa damage index were used to help selecting the right dose or ratio of decoctions for further study. Results The combined administration of Bletilla striata and Radix aconiti preparata at a certain proportion could reduce gastric ulcer index and stomach mucosa damage index. According to the regression analysis, the gastric ulcer index and stomach mucosa damage index were obviously inhibited by Bletilla striata, while this function decreased with the combined administration of Bletilla striata and Radix aconiti preparata. However, an interactive effect between each other has not been found. The influence of decoctions on the gastric ulcer index and stomach mucosa damage index became less when the compatibility ratio of Radix aconiti preparata increased, which referred to the total dose ranged between 3.64 g/kg and 29.32 g/kg. Conclusions The effect of the combined administration of Bletilla striata and Radix aconiti preparata showed relationship among the doses and ratios on anhydrous ethanol-induced gastric mucosal injury in mice with gastric ulcer.
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Objective To establish a molecular typing system of Staphylococcus aureus by using resolution melting for food-poi-soning fast tracing.Methods Primers were designed and synthesized according to the literature of VNTR in Staphylococcus au-reus ,and were used to perform molecular typing on the strains which had detected by PFGE,then 4 types of VNTRs were with higher discriminatory power were selected.On this basis,we established a molecular typing system for the detection of 59 Staphy-lococcus aureus isolated from food poisoning.Results The molecular typing system has good precision for detection.The standard deviation(s)of within-batch repetitive experiments were 0.03 -0.05 ℃,between-batch repetitive experiments were 0.04 -0.06℃,between-day repetitive experiments were 0.04-0.06 ℃.At the same time,the 59 strains of Staphylococcus aureus were divided into 19 types which were 11 epidemic clones and 8 sporadic clones.The correlation coefficient of Simpson was 0.916 4.Conclusion The molecular typing system for Staphylococcus aureus based on resolution melting was simple,fast and repeatable.It can be ap-plied to fast tracing and screen of Staphylococcus aureus in food poisoning.