Koumine regulates macrophage M1/M2 polarization via TSPO, alleviating sepsis-associated liver injury in mice.

BACKGROUND: Translocator protein (TSPO) is an 18-kDa transmembrane protein found primarily in the mitochondrial outer membrane, and it is implicated in inflammatory responses, such as cytokine release. Koumine (KM) is an indole alkaloid extracted from Gelsemium elegans Benth. It has been reported to be a high-affinity ligand of TSPO and to exert anti-inflammatory and immunomodulatory effects in our recent studies. However, the protective effect of KM on sepsis-associated liver injury (SALI) and its mechanisms are unknown. PURPOSE: To explore the role of TSPO in SALI and then further explore the protective effect and mechanism of KM on SALI. METHODS: The effect of KM on the survival rate of septic mice was confirmed in mouse models of caecal ligation and puncture (CLP)-induced and lipopolysaccharide (LPS)-induced sepsis. The protective effect of KM on CLP-induced SALI was comprehensively evaluated by observing the morphology of the mouse liver and measuring liver injury markers. The serum cytokine content was detected in mice by flow cytometry. Macrophage polarization in the liver was examined using western blotting. TSPO knockout mice were used to explore the role of TSPO in sepsis liver injury and verify the protective effect of KM on sepsis liver injury through TSPO. RESULTS: KM significantly improved the survival rate of both LPS- and CLP-induced sepsis in mice. KM has a significant liver protective effect on CLP-induced sepsis in mice. KM treatment ameliorated liver ischaemia, improved liver pathological injuries, and decreased the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH) and proinflammatory cytokines in serum. Western blotting results showed that KM inhibited M1 polarization of macrophages and promoted M2 polarization. In TSPO knockout mice, we found that TSPO knockout can improve the survival rate of septic mice, ameliorate liver ischaemia, improve liver pathological injuries, and decrease the levels of ALT, AST, and LDH. In addition, TSPO knockout inhibits the M1 polarization of macrophages in the liver of septic mice and promotes M2 polarization and the serum levels of proinflammatory cytokines. Interestingly, in TSPO knockout septic mice, these protective effects of KM were no longer effective. CONCLUSIONS: We report for the first time that TSPO plays a critical role in sepsis-associated liver injury by regulating the polarization of liver macrophages and reducing the inflammatory response. KM, a TSPO ligand, is a potentially desirable candidate for the treatment of SALI that may regulate macrophage M1/M2 polarization through TSPO in the liver.

Collective Plasmon Coupling in Gold Nanoparticle Clusters for Highly Efficient Photothermal Therapy.

Plasmonic nanomaterials with strong absorption at near-infrared frequencies are promising photothermal therapy agents (PTAs). The pursuit of high photothermal conversion efficiency has been the central focus of this research field. Here, we report the development of plasmonic nanoparticle clusters (PNCs) as highly efficient PTAs and provide a semiquantitative approach for calculating their resonant frequency and absorption efficiency by combining the effective medium approximation (EMA) theory and full-wave electrodynamic simulations. Guided by the theoretical prediction, we further develop a universal strategy of space-confined seeded growth to prepare various PNCs. Under optimized growth conditions, we achieve a record photothermal conversion efficiency of up to ∼84% for gold-based PNCs, which is attributed to the collective plasmon-coupling-induced near-unity absorption efficiency. We further demonstrate the extraordinary photothermal therapy performance of the optimized PNCs in in vivo application. Our work demonstrates the high feasibility and efficacy of PNCs as nanoscale PTAs.

Astragaloside IV promotes the angiogenic capacity of adipose-derived mesenchymal stem cells in a hindlimb ischemia model by FAK phosphorylation via CXCR2.

BACKGROUND: Therapeutic angiogenesis by transplantation of autologous/allogeneic adipose stem cells (ADSCs) is a potential method for the treatment of critical limb ischemia (CLI). However, the therapeutic efficiency is limited by poor viability, adhesion, migration and differentiation after cell transplantation into the target area. Astragaloside IV (AS-IV), one of the main active components of Astragalus, has been widely used in the treatment of ischemic diseases and can promote cell proliferation and angiogenesis. However, there is no report on the effect of AS-IV on ADSCs and its effect on hindlimb ischemia through cell transplantation. PURPOSE: The purpose of this study was to elucidate that AS-IV pretreatment enhances the therapeutic effect of ADSC on critical limb ischemia, and to characterize the underlying molecular mechanisms. METHODS: ADSCs were obtained and pretreated with the different concentration of AS-IV. In vitro, we analyzed the influence of AS-IV on ADSC proliferation, migration, angiogenesis and recruitment of human umbilical vein endothelial cells (HUVECs) and analyzed the relevant molecular mechanism. In vivo, we injected drug-pretreated ADSCs into a Matrigel or hindlimb ischemia model and evaluated the therapeutic effect by the laser Doppler perfusion index, immunofluorescence, and histopathology. RESULTS: In vitro experiments showed that AS-IV improved ADSC migration, angiogenesis and endothelial recruitment. The molecular mechanism may be related to the upregulation of CXC receptor 2 (CXCR2) to promote the phosphorylation of focal adhesion kinase (FAK). In vivo, Matrigel plug assay showed that ADSCs pretreated with AS-IV have stronger angiogenic potential. The laser Doppler perfusion index of the hindlimbs of mice in the ADSC/AS-IV group was significantly higher than the laser Doppler perfusion index of the hindlimbs of mice of the ADSC group and the control group, and the microvessel density was significantly increased. CONCLUSION: Our results demonstrate that AS-IV pretreatment of ADSC improves their therapeutic efficacy in ameliorating severe limb exclusion symptomology through CXCR2 induced FAK phosphorylation, which will bring new insights into the treatment of severe limb ischemia.

[Multi-residue determination of 289 pesticides in garlic by gas chromatography and gas chromatography/mass spectrometry].

A new idea of solvent transfer technique was developed and applied to determine 283 pesticide residues in garlic by gas chromatography-mass spectrometry (GC-MS) (method I), and the other method using normal phase silica/selective elution technique was applied to determine 6 pesticide residues with strongly polar in garlic by gas chromatography (method II). For the method I, the residues were extracted from homogenized tissue with acetonitrile-water, separated with liquid-liquid partition; the clear supernatant was purified by solvent transfer technique and solid phase extraction (Envi-18 and LC-NH2 columns), then was analyzed by GC-MS. For the method II, the residues were extracted from homogenized tissue using ethyl acetate and sodium sulfate assisted by ultrasonication. The supernatant was purified by solid phase extraction (primary secondary amine (PSA) powder and LC-Si column) prior to GC analysis. The determination was performed by using selected ion monitoring (SIM) mode in GC-MS method and flame photometric detector (FPD) in GC method, then external standard method was used in the quantification. Under the optimal conditions, the detection limits for the two methods (S/N > or = 10) of pesticides were 0.01-0.05 mg/kg, the recoveries carried out by the addition of standards of 0.01-0.20 mg/kg were 52%-163%, among which the recoveries for 88% pesticides were between 70% and 120%; the recoveries of the method II were 70%-111%; while the relative standard deviations were 2.4%-18% and 3.2%-9.3%, respectively. The model of solvent transfer technique and the sensitivity improvement of GC-MS was also studied. The methods are easy, fast, more sensitive, and can meet the requirement of the multiresidual analysis in garlic.

VKORC1 haplotypes influence the performance characteristics of PIVKAII for screening of hepatocellular carcinoma.

BACKGROUND: PIVKAII (protein induced by vitamin K absence), used for screening for hepatocellular carcinoma (HCC), is influenced by vitamin K epoxide reductase complex subunit 1 (VKORC1). VKORC1 haplotype frequency is significantly different in ethnic groups. We evaluated whether VKORC1 haplotypes could influence the performance characteristics of PIVKAII in screening for HCC. METHODS: A total 228 HCC patients and 258 patients with hepatitis B were recruited. Tumor size was measured in 76 patients with HCC. Serum PIVKAII concentrations and VKORC1 haplotype were determined in the cohort. Youden's index and ROC curves were used to compare the performance characteristics of PIVKAII in screening for HCC. RESULTS: In the HCC group and in patients with hepatitis B, serum PIVKAII concentrations were higher in VKORC1 rs2395612 TT carriers than in CC/CT carriers (50.34±72.18 vs. 11.98±27.45, p<0.05 in HCC group and 1.92±0.52 vs. 1.48±0.36, p<0.01). The estimated optimal cut-off value of PIVKAII for screening HCC was 2.0 and 3.0 ng/mL in CC/CT carriers and TT carriers, respectively. Furthermore, VKORC1 haplotypes also influenced the association of serum PIVKAII concentrations with HCC tumor size in patients with HCC. CONCLUSIONS: VKORC1 haplotypes influence the performance characteristics of PIVKAII for screening of HCC. Thus, measurement could be complementary for PIVKAII in HCC screening.

[Protective effect of Danzhi-xiaoyao San on rat brain energy or material metabolism (correction of matebolism) dealt with D-galactose].

OBJECTIVE: To research the mechanism of Danzhi-xiaoyao San (DZXYS) for treating Alzheimer's disease model of rats dealt with D-galactose. METHOD: An Alzheimer's disease-like model of rats has been set up with sc. D-galactose 150.0 mg kg-1 D-1 x 49 d. Comparing with Acricept in 0.54 mg kg-1 D-1 dosage as a positive control drug, DZXYS in 12.636 g kg-1 D-1 x 49 d dosage has orally been administrated orally to treat the injury in the Alzheimer's disease-like model of rats. The energy charge in the cerebral tissues had been detested with waters liquid chromatography; the protein content and DNA content in the cerebral tissues had been detested with ultraviolet assay, the relative content of aldose reductase-mRNA is detested with RT-PCR. The difference was analyzed between the control rats without D-galactose, the model rats dealt with D-galactose, the model rats treated with Aricept and the model rats treated with DZXYS, it is significant as P<0.05. RESULT: 1) DZXYS can not affect the energy charge in their cerebral tissues. 2) DZXYS can increase the protein content from 0.3139 +/- 0.019468 to 0.3213 +/- 0.015528 (ni=10, P>0.05) in their cerebral tissues. 3) DZXYS can increase the total DNA content from 1.093 +/- 0.267 to 1.488 +/- 0.341 (ni=10, P<0.01) in their cerebral tissues. 4) DZXYS can increase the content of AR-mRNA in their cerebral tissue from 0.732 +/- 0.159 to 1.418 +/- 0.277 (ni=5, P<0.01). CONCLUSION: It suggests that DZXYS could be effective in human Alzheimer's disease for its stabling gene expression, maintaining protein characteristics, recovering signal transduction in the Alzheimer's disease-like model rats dealt with D-galactose.

[Study on cardiac gene expression microarray and recipe for activating blood circulation and supplementing qi on heart of rats with post-infarction heart failure].

OBJECTIVE: To observe the effect of recipe for activating blood circulation and supplementing Qi (RAS) on cardiac functional structure in rats with post-infarction heart failure (PIHF). METHODS: Rat model of PIHF was established by left coronary artery ligation. Left ventricular samples of model rats from infarcted or peri-infarcted area were obtained at PIHF formation stage and stable stage (10 days and 8 weeks respectively after operation), the total RNA extracted and detected using 6 pieces of rat's 40s gene microarray (4096 genes/microarray), the data were analyzed using software as Genespring, Treeview, Clustering and SOM. Besides, RAS was used to treat the model rats beginning from 4 weeks after modeling and lasted for 4 weeks, changes of heart function and cardiac coefficient before and after treatment were observed by impedance method with Captopril as positive control. RESULTS: (1) Genespring analysis showed thousands of genes differential expression (upper or down regulated), including 13 kinds of gene involving energy metabolism, myocardial cytoskeleton, fibrosis, etc. which, in the infarcted area at heart formation stage were 1086 genes and at the stable stage, 724 genes, while in the peri-infarcted area, formation stage 196 genes and stable stage 97 genes. (2) After RAS or Captopril treatment, the heart function improved significantly, with the stroke volume, cardiac output and cardiac index increased significantly (P < 0.01). RAS could also improve the cardiac coefficient of model rats, as compared with that in untreated model, P < 0.01, compared with that in the sham-operated rats, P < 0.05. CONCLUSION: PIHF is a kind of overload heart disease with multiple genes abnormality. RAS could improve the heart function and histologic indexes, so as to treat the heart failure.

[Effect of different drug dosage to activate blood circulation and to nourish qi on cardiac function and structure of congestive heart failure rats after acute myocardial infarction].

OBJECTIVE: To observe the effect of different drug dosage to activate blood circulation and to nourish Qi on cardiac function and structure of heart failure rats after acute myocardial infarction. METHOD: The congestive heart failure post cardiac infarction rats models subjected to left coronary occlusion were made and given different dosage of drug of Huoxue Yiqi and captopril (positive control) from 4 weeks to 8 weeks after operation. The cardiac function before and after using drugs were observed by impedance methods. RESULT: Treated by different dosage of drug of Huoxue Yiqi and captopil, cardiac function of heart failure rats after acute myocardial infarction improved, SV, CO, CI(P < 0.01); at the same time, prescription of Huoxue Yiqi and Qixue could improve the model rats' heart coefficient, VS SHAM(P > 0.05), but prescription of Yiqi Huoxue and captopril could not improve it. CONCLUSION: Drug to activate blood circulation and to increase qi can treat heart failure rats after acute myocardial infarction, through improving their function and structure. At the same time, more dosage of drug to activate blood circulation and to increase qi(Huoxue/Yiqi drug) may improve the model rats' heart coefficient.