Effects of Yangyin Huoxue Prescription on the imbalance of Treg/Th17 and atherosclerosis in ApoE-/- mice induced by high-fat diet / 中国药科大学学报

@#Atherosclerosis(AS), characterized with the accumulation of lipids on the vessel wall, is an immune-related inflammatory disease which promotes the progression of cardiovascular diseases(CVD). The imbalance of Treg/Th17 accelerates the progression of AS. Yangyin Huoxue Prescription(YHF)is an efficient traditional Chinese medicine used in the treatment of AS, but the effects of YHF on the balance of immunity have still not been clarified. This project was designed to investigate the effects of YHF on the imbalance of Treg/Th17 and AS in ApoE-/- mice induced by high-fat diet(HFD). ApoE-/- mice were given HFD to induce AS and administered low-dose YHF(18 g/kg)or high-dose YHF(36 g/kg)for 20 weeks. Atherosclerotic plaque area was analyzed by oil red O staining. Serum lipids were measured by biochemical kits. Treg or Th17 cells in peripheral blood were detected by flow cytometry. mRNA and protein expression of Foxp3 and RORγt of aortas were determined by qRT-PCR, Western blot and immunohistochemistry. Splenic CD4+T cells of mice were isolated and activated by anti-CD3/CD28, and then treated with lipopolysaccharide(LPS)and YHF. The expression of mRNA and protein of Foxp3 and RORγt were detected by qRT-PCR and immunofluorescence. It was found that YHF reduced the plaque area, decreased lipid level and increased the ratio of Treg cells in peripheral blood. Moreover, YHF increased mRNA or protein expression of Foxp3 in aortas in vivo or CD4+T cells in vitro while decreasing mRNA or protein expression of RORγt. These results suggested that YHF can regulate the imbalance of Treg/Th17 in ApoE-/- mice induced by HFD, and reduce the inflammatory stimulation of LPS on CD4+T cells, thereby improving AS.

Effects of Mycobacterium tuberculosis HSP65 on Treg/Th17 immune balance in ApoE-knockout mice / 中国药科大学学报

@#To investigate the effects of Mycobacterium tuberculosis heat shock protein 65(HSP65)on Treg/Th17 immune balance in ApoE-knockout(ApoE-/-)mice, ApoE-/- mice with a high-cholesterol diet were immunized with M. tuberculosis HSP65. Sera were obtained for measurement of anti-HSP65 antibodies by ELISA; the effect of administration of different antigens was investigated, respectively, using flow cytometry analysis on the number of CD4+CD25+Foxp3+Tregs and CD4+IL-17+ Th17; the production of cytokines(IL-10, TGF-β1, IL-17 and IL-21)by these cells were determined by ELISA; total plasma cholesterol(TC), triglyceride(TG), high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)levels were detected by biochemical autoanalyzer. Atherosclerotic lesions were measured by lipid deposition stained with oil red O. The results demonstrated that the levels of anti-HSP65 IgG antibodies were increased significantly in Mycobacterium tuberculosis HSP65-treated ApoE-/- mice, revealed obvious decrease in Treg number, Treg related cytokines(IL-10, TGF-β1)levels and significant increase in Th17 number, Th17 related cytokines(IL-17 and IL-21)levels, the levels of TC, TG, HDL-C and LDL-C did not change between groups, while the atherosclerotic lesions significantly increased. Results indicate that M. tuberculosis HSP65 could interrupt the Th17/Treg immune balance in ApoE-/- mice, suggesting a potential role in the formation and progression of atherosclerosis.

Effects of nasal immunization of multi-target preventive vaccines on atherosclerosis.

Previous investigations have demonstrated that anti-inflammatory or lipid-lowering treatments could be useful for alleviating morbidity and mortality of atherosclerotic cardiovascular diseases. However, whether a vaccine designed to target inflammation and lipid simultaneously is more powerful to control the process of atherosclerosis remain to be unknown. Here, a vaccine was designed to target heat shock protein-65(Hsp65) and cholesteryl ester transfer protein (CETP) simultaneously and the effects of nasal immunization of multi-target vaccine on high-cholesterol-diet-driven rabbit atherosclerosis lesions were evaluated. Sera, nasal lavages and lung washes were used to ELISA assay for the analysis of IgG and IgA against Hsp65 and CETP. Sera were also used to the analysis of the avidity of combination of anti-Hsp65 and anti-CETP IgG antibodies with corresponding antigen, cytokines IL-10 and IFN-γ, and lipoproteins. In addition, aortas were harvested for analysis of atherosclerotic lesions. The results showed that lower and lasting specific anti-Hsp65 IgG and high anti-CETP IgG in sera and protective anti-Hsp65 and anti-CETP IgA in nasal cavity and lung were induced, the avidity of combination of anti-Hsp65 and anti-CETP IgG with antigen were higher, and more protective IL-10 and less adverse IFN-γ were produced. In addition, sera TC, and LDL-C were decreased. As a result, the size of aorta atherosclerotic plaques was significantly reduced. We conclude that multifaceted vaccine combining lipid-regulating with anti-inflammation was a potential remedy, especially for atherosclerosis with complicated etiology.

Shikonin inhibits the chemotaxis of monocytes in endometriosis via regulating RANTES expression / 中国免疫学杂志

Objective:To evaluate the effect of shikonin on the expression of RANTES and chemotactic activity of monocyte in endometriosis.Methods:Established SCID endometriosis models and cultured U937 cells were treated by a series of concentration of shikonin.RANTES transcriptive expression was determined by Real-time PCR,and RANTES secretion was determined by ELISA.Furthermore,Chemotaxis assay in vitro was conducted to elucidate the effect of shikonin on chemotaxis of U937 cells by RANTES.Results:Shikonin improved the RANTES transcription of human endometrium transplanted to SCID mouse(P

Protective Mechanism of Salvianolic Acid B on Myocardial Ischemia-reperfusion Injury of Rats / 中药新药与临床药理

Objective To investigate the protective mechanism of Salvianolic acid B (Sal B) on experimental in-vivo myocardial ischemia-reperfusion injury (MIRI)in rats. Methods Rats model of myocardial ischemia-reperfusion injury was induced by ischemia for 60 min and then reperfusion for 60 min. After treatment,endothelin (ET) levels in plasma and the homogenate of myocardial tissue,and serum levels of nitric oxide (NO) and NO synthase were measured. Meanwhile,the pathological changes of myocardial tissue were also observed. Results Compared with the model group,middle-dose and high-dose Sal B could depress the ET levels in the plasma and the myocardial tissue,and increase the content of serum NO and NOS (P

Effect of Shuluo Injection on Cerebral Ischemia-reperfusion in Mice / 中药新药与临床药理

Objective To investigate the effect of Shuluo Injection on cerebral water volume,cerebral index ,LDH,NOS and SOD in the brain.Methods The model of cerebral ischemia-reperfusion in mice is set up by separating the bilateral common carotid artery (CCA) of the mice,blocking the blood stream by straining the thread and by reperfusion loosing the thread. After 3 times of ischemia-reperfusion,brain samples were prepared to observe the cerebral injury. Results High-dose Shuluo Injection significantly reduced the cerebral water volume and decreased the cerebral index and the activity of LDH and NOS(P