miR-212-3p attenuates neuroinflammation of rats with Alzheimer's disease via regulating the SP1/BACE1/NLRP3/Caspase-1 signaling pathway.

Alzheimer's disease (AD) ranks as the leading cause of dementia. MicroRNA (miR)-212-3p has been identified to exert neuroprotective effects on brain disorders. The current study analyzed the protective role of miR-212-3p in AD rats via regulating the nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3)/Caspase-1 signaling pathway. The AD rat model was established via injection of amyloid-ß 1-42 (Aß1-42), followed by the Morris water maze test. The morphology and functions of neurons were observed. Furthermore, miR-212-3p, NLRP3, cleaved Caspase-1, gasdermin D N-terminus, interleukin (IL)-1ß and IL-18 expressions were measured. H19-7 cells were treated with Aß1-42 to establish the AD cell model, followed by an assessment of cell viability and pyroptosis. Downstream targets of miR-212-3p and specificity protein 1 (SP1), as well as beta-site amyloid precursor protein cleaving enzyme 1 (BACE1) were predicted by databases and testified using dual-luciferase and chromatin immunoprecipitation assays. miR-212-3p was weakly expressed in AD rats. miR-212-3p overexpression was linked to improved learning and memory capacities of AD rats and reduced neuronal pyroptosis linked to neuroinflammation attenuation. In vitro, miR-212-3p improved viability and suppressed pyroptosis of neurons via inhibiting NLRP3/Caspase-1. Overall, miR-212-3p inhibited SP1 expression to block BACE1-induced activation of NLRP3/Caspase-1, thereby attenuating neuroinflammation of AD rats.

Mangiferin inhibits macrophage classical activation via downregulating interferon regulatory factor 5 expression.

Mangiferin is a natural polyphenol and the predominant effective component of Mangifera indica Linn. leaves. For hundreds of years, Mangifera indica Linn. leaf has been used as an ingredient in numerous traditional Chinese medicine preparations for the treatment of bronchitis. However, the pharmacological mechanism of mangiferin in the treatment of bronchitis remains to be elucidated. Macrophage classical activation is important role in the process of bronchial airway inflammation, and interferon regulatory factor 5 (IRF5) has been identified as a key regulatory factor for macrophage classical activation. The present study used the THP­1 human monocyte cell line to investigate whether mangiferin inhibits macrophage classical activation via suppressing IRF5 expression in vitro. THP­1 cells were differentiated to macrophages by phorbol 12­myristate 13­acetate. Macrophages were polarized to M1 macrophages following stimulation with lipopolysaccharide (LPS)/interferon­Î³ (IFN­Î³). Flow cytometric analysis was conducted to detect the M1 macrophages. Reverse transcription­quantitative polymerase chain reaction was used to investigate cellular IRF5 gene expression. Levels of proinflammatory cytokines and IRF5 were assessed following cell culture and cellular homogenization using enzyme­linked immunosorbent assay. IRF5 protein and nuclei co­localization was performed in macrophages with laser scanning confocal microscope immunofluorescence analysis. The results of the present study demonstrated that mangiferin significantly inhibits LPS/IFN­Î³ stimulation­induced classical activation of macrophages in vitro and markedly decreases proinflammatory cytokine release. In addition, cellular IRF5 expression was markedly downregulated. These results suggest that the inhibitory effect of mangiferin on classical activation of macrophages may be exerted via downregulation of cellular IRF5 expression levels.

[Study on the regulation role of semen persicae to cAMP-PKA signal pathway in the rats with cold and heat blood stasis syndrome].

OBJECTIVE: To study the regulation role of with neutral property to cAMP-PKA pathway in the rats with cold and heat blood stasis syndromes and it's mechanism. METHODS: 60 rats were randomly divided into normal control group, model control group, Semen Persicae group, radix salvia miltiorrhiza group, rhizoma chuanxiong group, 12 rats per group. The three herb groups were orally given relative herbs decoction, whose dosages were equal to 10 times the human clinical dose, normal and model control groups were orally given water, 2 times/day, 20 mL/kg, for 7 days. Experiments in rats with cold and heat blood stasis syndromes were carried on respectiverly. In heat blood stasis syndromes, except normal control group, the other groups were intraperitoneally injected 10% carrageenan, 5 mL/kg, 1 times/day, for 3 days;24 hours after the last injection, subcutaneously injected 20% dry yeast suspension, 10 mL/kg. In cold blood stasis syndromes, except normal control group, the other groups were put into fridge, temperature--(18 +/- 2) degrees C, 2 hours/ times, 2 times/day, for 7 days. Separately draw 5 ml abdominal aortic blood and taken abdominal aorta, 6 hours and 12 hours after finishing the model in the two syndromes. Tested the cAMP content by elisa, tested the PKA protein expression by Western blot. RESULTS: Semen Persicae with neutral property, could decrease the content of cAMP in plasma (P < 0.01), inhibit the expression of protein PKA (P < 0.05) in rats with heat stagnation and blood stasis syndrome, increase the plasma content of cAMP (P < 0.01) and increase the expression of protein PKA (P < 0.01) in rats with cold stagnation and blood stasis syndrome. Semen Persicae had two-way adjustment action on CAMP-PKA signal pathway. CONCLUSION: In different internal environment of heat stagnation and blood stasis syndrome, cold stagnation and blood stasis syndrome, Semen Persicae with neutral property has two-way adjustment to cAMP-PKA signaling channel, which may be one of the mechanism of it's two-way application.

[Experimental study on two-way application of drugs with neutral property for promoting blood circulation and removing blood stasis on cold and heat blood stasis syndromes II].

OBJECTIVE: To further study the characteristics of drugs with neutral property in two-way application and conditioned dominance by observing the action characteristic of 10 traditional Chinese medicines with neutral property in hemorheological indicators of heat stagnation and blood stasis syndrome and cold stagnation and blood stasis syndrome rats. METHOD: The model of heat stagnation and blood stasis syndrome rats was established by injecting carrageenan and dry yeast, while the model of cold stagnation and blood stasis syndrome rats was established by body freezing. Subsequently, 10 traditional Chinese medicines with neutral property, 5 traditional Chinese medicines with heat property and 5 traditional Chinese medicines with cold property were selected for intervention to observe the changes in such indicators as whole blood viscosity, plasma viscosity and hematocrit and analyze the action characteristics of drugs with neutral property. RESULT: ANOVA showed that among six of the 10 traditional Chinese medicines with neutral property, including Typhae Pollen, Sarcandrae Herba and Sappan lignum, could obviously increase the hemorheological indicators of both heat stagnation and blood stasis syndrome and cold stagnation and blood stasis syndrome rats; five traditional Chinese medicines with cold property, such as Salviae Miltiorrhizae Radix et Rhizoma, Leonuri Herba, Rhei Radix et Rhizoma, could significantly ameliorate the hemorheological indicators of heat stagnation and blood stasis syndrome rats (P < 0.01 or P < 0.05), and Salvia Miltiorrhiza Radix et Rhizoma alone could ameliorate the hemorheological indicators of cold stagnation and blood stasis syndrome rats (P < 0.05); all of the five traditional Chinese medicines with heat property could significantly ameliorate the hemorheological indicators of cold stagnation and blood stasis syndrome rats (P < 0.01), among which Carthami Flos and Notoginseng Radix et Rhizoma could significantly ameliorate the hemorheological indicators of cold stagnation and blood stasis syndrome rats. According to the average high-shear blood viscosity analysis, drugs with neutral property showed similar action characteristics to those with cold property in ameliorating hemorheology indicators of heat stagnation and blood stasis syndrome rat and better effect than those with heat property in reducing whole blood viscosity; and traditional Chinese medicines with neutral property have the similar action characteristics to those with heat property in improving the hemorheology indicators of cold stagnation and blood stasis syndrome rat and better effect than those with heat property in reducing whole blood viscosity. CONCLUSION: Under the condition of heat stagnation and blood stasis syndrome, traditional Chinese medicines with neutral property show the similar action characteristics to those with cold property; but under the condition of cold stagnation and blood stasis syndrome, traditional Chinese medicines with neutral property show the similar action characteristics to those with heat property. This indicates that traditional Chinese medicines with neutral property show both heat and cold properties under he conditions of heat stagnation and blood stasis syndrome and cold stagnation and blood stasis syndrome.

[Experimental study on two-way application of traditional Chinese medicines capable of promoting blood circulation and removing blood stasis with neutral property in cold and hot blood stasis syndrome I].

OBJECTIVE: To study the action characteristics of "two-way application and conditioned dominance" of traditional Chinese medicines with neutral property by observing the action characteristic of 10 traditional Chinese medicines capable of promoting blood circulation and removing blood stasis with neutral property in the microcirculation in rats with heat stagnation and blood stasis syndrome. METHOD: The rat model with heat stagnation and blood stasis syndrome was established by injecting carrageenan and dry yeast, and the rat model with cold stagnation and blood stasis syndrome was built by the body freezing method. Ten traditional Chinese medicines with neutral property, including 5 with hot property and 5 with cold property, were selected for intervention to observe blood flow rate and flow state indicators in rat auricles and make a comparative analysis on action characteristics of traditional Chinese medicines with neutral property. RESULT: ANOVA showed that among the 10 traditional Chinese medicines with neutral property, 6 such as Typhae Pollen, Sappan Lignum and Vaccariae Semen can obviously increase the blood flow rate (P < 0.01 or P < 0.05) in the above two models; all of the 5 traditional Chinese medicines with cold property can increase the blood flow rate (P < 0.01 or P < 0.05) in the rat model with heat stagnation and blood stasis syndrome, but only Salvia miltiorrhiza can increase the blood flow rate (P < 0.01 or P < 0.05) in the rat models with cold stagnation and blood stasis syndrome, while other medicines showed no notable effect; among the 5 traditional Chinese medicines with hot property, Carthamus tinctorius and Ligusticum chuanxiong can increase the blood flow rate (P < 0.01 or P < 0.05) in the rat models with cold stagnation and blood stasis syndrome, but had no obvious effect to the blood flow rate in the rat models with heat stagnation and blood stasis syndrome. According to the analysis on average blood flow rate, traditional Chinese medicines with natural and cold properties showed similar effect on heat stagnation and blood stasis syndrome and better effect in increasing blood flow rate than those with hot property; those with natural and hot properties showed similar effect and better effect in increasing blood flow rate than those with cold property. CONCLUSION: Under the condition of heat stagnation and blood stasis syndrome, traditional Chinese medicines with neutral property have the similar action characteristics with those with cold property; wile under the condition of cold stagnation and blood stasis syndrome, traditional Chinese medicines with neutral property have the similar action characteristics with the Chinese medicinal herbs with hot property. This indicates the action characteristics of "two-way application and conditioned dominance" of traditional Chinese medicines with neutral property to some extent.

Amygdalin mediates relieved atherosclerosis in apolipoprotein E deficient mice through the induction of regulatory T cells.

OBJECTIVE: Regulatory T cells (Tregs) play a critical role in the regulation of T cell-mediated immune responses in atherosclerosis, a chronic autoimmune-like disease. Therefore, in this study, we aimed to investigate the therapeutic effect of amygdalin on atherosclerosis of apolipoprotein E deficient (ApoE(-/-)) mice, and to explore its immune regulatory function by stimulation of Tregs. METHODS AND RESULTS: To evaluate the anti-atherosclerotic effect of amygdalin and for in vivo Treg expansion/activation analysis, ApoE(-/-) mice received intraperitoneal injections of amygdalin, and this therapy resulted in a comparatively 2-fold decrease in triglyceride (TG), 1.5-fold decrease in total cholesterol (TC) and low density lipoprotein (LDL). By comparing the vessel areas, lumen areas, plaque areas, and aortic plaque coverage percentage, the effects of amygdalin on pre-existing lesions were assessed. Studies on IL-10 and TGF-ß indicated that mice treated with amygdalin had increased expression of Treg-related cytokines. Meanwhile, flow cytometry and real-time PCR data showed that mice treated with amygdalin had higher percentage of CD4(+)CD25(+)Foxp3(+) T cells than untreated mice and increased expression of forkhead box P3 (FOXP3) gene. CONCLUSION: Our data showed amygdalin could attenuate the development of atherosclerosis by suppressing inflammatory responses and promoting the immunomodulation function of Tregs. The effects of amygdalin ultimately resulted in the enlarged lumen area and the loss of atherosclerotic plaque. All these data indicated the therapeutic potential of amygdalin in preventing and/or treating of atherosclerosis.

[Naphtha analysis of different processed products of Rhizoma Curcumae from Guangxi by GC-MS].

OBJECTIVE: To approach the naphtha analysis of different processed products of Rhizoma Curcumae from Guangxi. METHODS: The naphtha was extracted by steam distillation. The relative content of every compound was determined with area normalization method and the structures were elucidated by GC-MS technique. RESULTS: 33 kinds of chemical compositions were identified in the naphtha of the six samples,the main contents were Cyclohexane, 1-ethenyl-1-methyl-2,4-bis (1-methylethenyl)-, 5-Benzofuranacetic acid, 6-ethenyl-4,5,6,7-terahydro-3, 6-dimethyl-alpha. -methylene-,methyl ester, and so on. CONCLUSIONS: Six samples of Rhizoma Curcumae are significantly different in quality,and the naphtha's content of the sample processing with vinegar is the highest.