Gallic acid promotes macrophage phagosome acidification and phagolysosome formation by activating NLRP3/mTOR signaling pathway.

INTRODUCTION: Gallic acid (GA) has a good therapeutic effect in bacteriological inhibition and plays a variety of functions in maintaining the stability of the immune system. The aim of the present study was to investigate the effect of GA on the bactericidal activity of macrophages against Vibrio vulnificus (Vv). METHODS: A cell counting kit-8 (CCK-8) assay was carried out to test the cytotoxicity of GA on J774A.1 cells. Concentration of proinflammatory cytokines in J774A.1 cells were evaluated by ELISA. The internalization and degradation of Vv in the phagosomes were observed by transmission electron microscopy (TEM). The phagosome acidification and phagolysosome formation were detected to evaluate the bacteria-clearing function of J774A.1 cells. The bactericidal activity of GA in vivo was also investigated by collecting the survival time of Vv infected mice and observing the inflammatory infiltration of organs. RESULTS: Our results demonstrated that GA at 50 µM significantly inhibited the proinflammatory cytokines levels, promoted phagosome acidification and phagolysosome formation in J774A.1 cells with Vv infection. This may be related to the activation of NLRP3/mTOR signaling pathway. Additionally, GA treatment improves the survival and bactericidal activity of mice infected with Vv. CONCLUSIONS: In summary, GA exerts bactericidal activity against Vv infection by regulating the formation and acidification of phagocytic lysosomes in macrophages.

Revealing active ingredients, potential targets, and action mechanism of Ermiao fang for treating endometritis based on network pharmacology strategy.

ETHNOPHARMACOLOGICAL RELEVANCE: Ermiao fang (EMF) is a traditional Chinese medicinal herbal formula from ancient times and recorded in the pharmacopeia of the People's Republic of China. It is composed of two typical Chinese herbal medicines, Cortex Phellodendri (Huangbai), the bark of Phellodendron chinensis Schneid. (Rutaceae), and Rhizoma Atractylodis (Cangzhu), the rhizome of Atractylodes lancea (Thunb.) DC. (Compositae). EMF has been clinically used for the treatment of endometritis for many years in China. AIM OF THE STUDY: This study was aimed to identify the active ingredients, potential targets, and mechanism of action of EMF for the treatment of endometritis. MATERIALS AND METHODS: In this research, the pharmacological effects of EMF on endometritis were first evaluated by establishing a rat model of endometritis. A network pharmacology-based analytical strategy was then used to predict its targets and signaling pathways. An endometritis-related protein target and compound database was built for EMF. The compounds in EMF and those absorbed into the blood were identified by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS). High-throughput virtual screening and molecule docking methods were used to predict the protein targets of EMF. The surface plasmon resonance analysis (SPR) method was used to validate the affinity between the compound and proteins. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was used to predict the related pathways. Western blotting analysis was used to evaluate the expression of key proteins in the related pathways. RESULTS: The animal study showed that EMF could reduce uterine inflammation in rats with endometritis. Then, an ingredient database including 187 compounds and a protein target database including 836 proteins were constructed. Twenty-four compounds in EMF were identified by UHPLC-Q-TOF/MS, among which eight compounds were present in rat plasma after an oral administration of EMF. Afterward, 39 potential target proteins were predicted by the high-throughput screening method, and 20 of them were selected after further screening using molecular docking. Subsequently, an ingredient-target network was constructed, and the target proteins were classified into the NF-κB and MAPK signal pathways by KEGG pathway enrichment analysis. Finally, the affinity between the active ingredients and the target proteins was verified by SPR. The Western blotting analysis showed that EMF significantly inhibited the elevated NF-κB and MAPK pathway proteins in rats with endometritis. CONCLUSIONS: EMF exhibited a significant pharmacological effect on rats with endometritis. Network pharmacology analysis revealed that eight compounds were absorbed into the blood after oral administration and interacted with 20 targets. Western blotting analysis indicated that EMF exerted anti-inflammatory effects by inhibiting the NF-κB and MAPK signaling pathway proteins in the treatment of endometritis.

[Study on extracting and separating curcuminoids from Curcuma longa rhizome using ultrasound strengthen by microemulsion].

OBJECTIVE: To establish a new method for the extraction and separation of curcuminoids from Curcuma longa rhizome by cloud-point preconcentration using microemulsions as solvent. METHODS: The spectrophotometry was used to detect the solubility of curcumin in different oil phase, emulsifier and auxiliary emulsifier, and the microemulsion prescription was used for false three-phase figure optimization. The extraction process was optimized by uniform experiment design. The curcuminoids were separated from microemulsion extract by cloud-point preconcentration. RESULTS: Oil phase was oleic acid ethyl ester; Emulsifier was OP emulsifier; Auxiliary emulsifier was polyethylene glycol(peg) 400; The quantity of emulsifier to auxiliary emulsifier was the ratio of 5: 1; Microemulsion prescription was water-oleic acid ethyl ester-mixed emulsifier (0.45:0.1:0.45). The optimum extraction process was: time for 12.5 min, temperature of 52 degrees C, power of 360 W, frequency of 400 kHz, and the liquid-solid ratio of 40:1. The extraction rate of curcuminoids was 92.17% and 86.85% in microemulsion and oil phase, respectively. CONCLUSION: Curcuminoids is soluble in this microemulsion prescription with good extraction rate. This method is simple and suitable for curcuminoids extraction from Curcuma longa rhizome.

[The common and variation peak ratio dual index sequence analysis on UV fingerprint spectra of Paeonia Lactiflora Pall].

Baishao and Chishao (Paeonia Lactiflora Pall.) and their close relative Danpi (Paeonia Suffruticosa Andr) samples were estimated quantitatively, based on their UV fingerprint spectra of the extracts obtained with chloroform, ethanol and water, by applying the common and variation peak ratio dual index sequence analysis method. The analytical results showed that the Baishao samples B2, B3 and B4 from the closest regions were the most similar samples. Their common peak ratios were larger than 70 percent and their variation peak ratios were less than 33.3 percent. However, there existed obvious differences among Baishao sample group 1 (B1 and B5), group 2 (B2, B3 and B4) and group 3 (B6) from different regions. The common peak ratios among group 1 (B1 and B5) and group 2(B2, B3 and B4) were lower than 60 percent, while those among group 1 (B1 and B5) and group 3 (B6) were less than 57 percent. The Baishao samples B1 and B5 from the same region collected in different years were of significant disparity, their common peak ratio was only 44.4 percent, but their variation peak ratios were larger than 100 percent. In fact, this method reaches the limitation of quantitative identification of herbs, and can distinguish at least two samples quantitatively.