ABSTRACT
Objective @#To fabricate a co-delivery system of curcumin (CUR) and siRNA based on mesoporous silica nanoparticles (MSN) and investigate its potential application in inducing macrophage M2 polarization.@*Methods@# MSNs were synthesized using the conventional sol-gel method. The interior mesochannels were occupied by small-molecule CUR, while the exterior surface was adsorbed by cationic polymeric polyethyleneimine (PEI) to link the negatively charged siRNA molecules to formulate the (CUR@MSN)PEI/siRNA co-delivery system. The formulation process was monitored by transmission electron microscopy(TEM). The MTT assay was used to evaluate the cytotoxicity in RAW264.7 cells under various concentrations of nanoparticles. Confocal laser scanning microscopy and TEM were used to observe cell internalization using FAM-labeled siRNA. GAPDH-targeting siRNA was used to prepare nanoparticles and then was transfected into RAW264.7 cells to observe the silencing efficiency of target genes. The knockdown efficiency was examined by real-time quantitative PCR. The related control groups were untreated cells, CUR delivery only and the co-delivery of CUR and siRNA negative control. By loading miRNA-130a-3p antisense oligonucleotide (ASO) to transfect RAW264.7 cells, the effects on the polarization of macrophages were observed. The M2 polarization marker arginase 1 (Arg-1) was measured by western blotting. The related control groups were untreated cells, CUR delivery only and co-delivery of CUR and miRNA negative control. @* Results @# The (CUR@MSN)PEI/siRNA co-delivery system was successfully formulated. The nanoparticles exhibited dose-dependent cytotoxicity, and the cell viability was maintained over 90% when the nanoparticle concentration was less than 10 μg/mL. A high cell uptake efficiency was observed, and the target gene knockdown efficiency was greater than 80% (P < 0.05 vs. all the other groups). The CUR delivery-only group and co-delivery of the CUR-and miRNA-negative control group improved Arg-1 expression ~ 3-fold (P < 0.05 vs. untreated cells). Using the co-delivery of CUR and ASO, synergistic effects were obtained, and Arg-1 expression was increased ~ 8-fold (P < 0.05 vs. all the other groups).@*Conclusion @#The CUR-siRNA co-delivery system can effectively transfect macrophages and synergistically induce M2 polarization.
ABSTRACT
The objective was to investigate the effect of kinsenoside (Kin) treatments on macrophage polarity and evaluate the resulting protection of chondrocytes to attenuate osteoarthritis (OA) progression. RAW264.7 macrophages were polarized to M1/M2 subtypes then administered with different concentrations of Kin. The polarization transitions were evaluated with quantitative real-time polymerase chain reaction (qRT-PCR), confocal observation and flow cytometry analysis. The mechanism of Kin repolarizing M1 macrophages was evaluated by Western blot. Further, macrophage conditioned medium (CM) and IL-1 were administered to chondrocytes. Micro-CT scanning and histological observations were conducted on anterior cruciate ligament transection (ACLT) mice with or without Kin treatment. We found that Kin repolarized M1 macrophages to the M2 phenotype. Mechanistically, Kin inhibited the phosphorylation of IB, which further reduced the downstream phosphorylation of P65 in nuclear factor-B (NF-B) signaling. Moreover, Kin inhibited mitogen-activated protein kinases (MAPK) signaling molecules p-JNK, p-ERK and p-P38. Additionally, Kin attenuated macrophage CM and IL-1-induced chondrocyte damage. , Kin reduced the infiltration of M1 macrophages, promoted M2 macrophages in the synovium, inhibited subchondral bone destruction and reduced articular cartilage damage induced by ACLT. All the results indicated that Kin is an effective therapeutic candidate for OA treatment.
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Objective To explore the activition and polarization of microglia in the epileptic rats induced by lithium chloride-pilocarpine. Methods One hundred male SD rats were randomly divided into five groups: control group and different time points model groups including 1d,3d,7d and 14d. Epilepsy models were established by lithium chloride-pilocarpine intraperitoneal injection. The control group was given the same dosage of normal saline. The morphology change was detected by immunofluorescence,and the expressions of iNOS and Arg-1 were determined by IHC at respective time points. Results Compared the model groups with control group,microglia was activated,synapsis was shorten,volume got bigger,most of them seemed as amoebocyte,the expression of iNOS increased and Arg-1 decreased,especially at 3d.ConclusionThe results from this study indicated that microglia was activated and polarized in epileptic rats induced by pilocarpine.
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Objective To investigate the effects of Schistosoma japonicum infection on selective activation of macrophages and insulin resistance in liver tissues of mice with high-fat diet induced obesity and the possible mechanism. Methods Thirty-six male C57BL/ 6J mice were randomly divided into 3 groups in-cluding normal control group(NC group,n=12),high-fat diet feeding group(HF group,n=12)and high-fat diet feeding with Schistosoma japonicum infection group(HSj group,n=12). Specimens were collected 6 and 12 weeks after high-fat diet feeding. The levels of fasting blood glucose(FBG),fasting plasma insulin (FINS)and homeostasis model assessment for insulin resistance index(HOMA-IR)were detected. The ex-pression of interleukin-6(IL-6),arginase-1(Arg-1)and found in inflammatory zone-1(Fizz-1)at mRNA and protein levels were detected by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR)and immunohistochemistry,respectively. Results The mice from HF group showed higher levels of HOMA-IR than those form NC group and HSj group by the end of 6 and 12 weeks after infection(P﹤0. 05). The levels of HOMA-IR in mice from HF group were increased by the end of week 12 after infection as com-pared with those at week 6(P>0. 05). The levels of IL-6 in mice from both HF group and HSj group were higher than those from NC group by the end of week 6 after infection(P﹤0. 05). Higher levels of IL-6 were detected in mice from HF group as compared with those from HSj group and NC group by the end of week 12 after infection(P﹤0. 05). The expression of Arg-1 and Fizz-1 in mice form HF group were lower than those from NC group by the end of 6 and 12 weeks after infection(P﹤0. 05). Arg-1 was highly expressed in mice form HSj group,followed by those from NC and HF groups 12 weeks after infection. The expression of Fizz-1 in mice from HSj group was the highest among the three groups by the end of week 6 and 12 after infection (P﹤0. 05). Conclusion The proinflammatory effects on mice with diet induced obesity were induced dur-ing acute infection of Schistosoma japonicum cercariae(6 weeks). The chronic infection of Schistosoma ja-ponicum cercariae(12 weeks)might be helpful in reversing hepatic insulin resistance in mice with diet in-duced obesity by changing the polarity of macrophages in liver tissues.
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Introducción: Variaciones moleculares en el gen de la arginasa I, ARGl, podrían provocar un aumento de la expresión de la enzima o de su actividad, lo que puede llevar a una disfunción endotelial al disminuir la producción de óxido nítrico por parte de la eNOS. Así, el objetivo del presente estudio fue investigar la posible asociación del polimorfismo rs2781666 G>T y la presencia de enfermedad coronaria (confirmada con angiografía) en individuos de la región de La Araucanía. Material y Métodos: En el presente estudio, de tipo casos y controles, fueron evaluados 247 sujetos, con edades entre 30 y 74 años; 124 individuos pacientes con enfermedad arterial coronaria (casos) y 123 controles. La genotipificación del polimorfismo rs2781666 del gen ARGl fue realizada mediante PCR- RFLR Resultados: La frecuencia del genotipo homocigoto TT para el polimorfismo rs2781666 del gen ARGl fue de 6.5 por ciento en el grupo casos y 8 por ciento en el grupo control, difiriendo significativamente (p=0.032). La frecuencia relativa del alelo T también presentó diferencias significativas entre casos y controles (0.230 vs. 0.325, p=0.031). La OR relacionada al alelo mutado T fue de 0.63 (I.C. 95 por ciento, 0.43 - 0.94), confirmando la presencia de la asociación observada. Conclusión: Los resultados obtenidos sugieren que el polimorfismo rs2781666 G>T del gen ARGl confiere protección contra enfermedad coronaria en la población analizada. Sin embargo, este resultado debe ser replicado en otros grupos poblacionales de nuestro país.
Background: Recent data support a role for arginase 1 (ARG1) in the initiation, development, and complications of coronary artery disease. Thus, in the present study we investigated the possible association between the rs2781666 G>T variant of ARG1 and the presence of CAD confirmed by angiography in Chilean subjects. Methods: A total of 124 unrelated patients with diagnosis of CAD confirmed by angiography (stenosis > 70 percent) and 123 healthy controls (30 - 74 years old) were included in this study. The rs2781666 G>T variant of the ARG1 gene was evaluated by PCR-RFLR Results: The frequency of TT homozygous genotype in CAD patients was lower when compared to control group (6.5 percent vs. 8.0 percent, p=0.032). Similarly, the T allele frequency was different between CAD and control groups (0.230 vs. 0.325, p=0.031). The OR for CAD related to T allele was 0.64 (95 percent CI. = 0.43-0.94), con-firming the association founded. Conclusion: These findings suggest that the T allele for rs2781666 polymorphism of the ARG1 gene constitutes an inherited protective factor against CAD in Southern Chilean subjects.