Reyanning mixture inhibits M1 macrophage polarization through the glycogen synthesis pathway to improve lipopolysaccharide-induced acute lung injury.

ETHNOPHARMACOLOGICAL RELEVANCE: Reyanning (RYN) mixture is a traditional Chinese medicine composed of Taraxacum, Polygonum cuspidatum, Scutellariae Barbatae and Patrinia villosa and is used for the treatment of acute respiratory system diseases with significant clinical efficacy. AIM OF THE STUDY: Acute lung injury (ALI) is a common clinical disease characterized by acute respiratory failure. This study was conducted to evaluate the therapeutic effects of RYN on ALI and to explore its mechanism of action. MATERIALS AND METHODS: Ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used to analyze the chemical components of RYN. 7.5 mg/kg LPS was administered to induce ALI in rats. RYN was administered by gavage at doses of 2 ml/kg, 4 ml/kg or 8 ml/kg every 8 h for a total of 6 doses. Observations included lung histomorphology, lung wet/dry (W/D) weight ratio, lung permeability index (LPI), HE staining, Wright-Giemsa staining. ELISA was performed to detect the levels of TNF-α, IL-6, IL-10, Arg-1,UDPG. Immunohistochemical staining detected IL-6, F4/80 expression. ROS, MDA, SOD, GSH/GSSG were detected in liver tissues. Multiple omics techniques were used to predict the potential mechanism of action of RYN, which was verified by in vivo closure experiments. Immunofluorescence staining detected the co-expression of CD86 and CD206, CD86 and P2Y14, CD86 and UGP2 in liver tissues. qRT-PCR detected the mRNA levels of UGP2, P2Y14 and STAT1, and immunoblotting detected the protein expression of UGP2, P2Y14, STAT1, p-STAT1. RESULTS: RYN was detected to contain 1366 metabolites, some of the metabolites with high levels have anti-inflammatory, antibacterial, antiviral and antioxidant properties. RYN (2, 4, and 8 ml/kg) exerted dose-dependent therapeutic effects on the ALI rats, by reducing inflammatory cell infiltration and oxidative stress damage, inhibiting CD86 expression, decreasing TNF-α and IL-6 levels, and increasing IL-10 and Arg-1 levels. Transcriptomics and proteomics showed that glucose metabolism provided the pathway for the anti-ALI properties of RYN and that RYN inhibited lung glycogen production and distribution. Immunofluorescence co-staining showed that RYN inhibited CD86 and UGP2 expressions. In vivo blocking experiments revealed that blocking glycogen synthesis reduced UDPG content, inhibited P2Y14 and CD86 expressions, decreased P2Y14 and STAT1 mRNA and protein expressions, reduced STAT1 protein phosphorylation expression, and had the same therapeutic effect as RYN. CONCLUSION: RYN inhibits M1 macrophage polarization to alleviate ALI. Blocking glycogen synthesis and inhibiting the UDPG/P2Y14/STAT1 signaling pathway may be its molecular mechanism.

The complete mitochondrial genome of Bullanga florida (Neuroptera: Myrmeleontidae).

The complete mitochondrial genome (mitogenome) of Bullanga florida (Neuroptera: Myrmeleontidae) was determined. The entire sequence is 15,937 bp in length which contains 13 protein-coding genes (PCGs), 22 tRNAs, two rRNAs and one non-coding AT-rich region. The overall A + T content of mitogenome is 73.3%. The gene order and arrangement are similar to other Neuroptera mitogenomes. Thirteen PCGs start with standard ATN initiation codons and stop with termination codons TAA or T. All tRNA genes have a typical clover-leaf secondary structure except for tRNASer (AGN), whose dihydrouridine (DHU) arm do not form a stable stem-loop structure. Two rRNA genes (rrnL and rrnS) are 1325 bp and 776 bp in size, respectively. The phylogenetic analysis is based on the amino acid sequences of 13 PCGs indicates that B. florida is a sister group to Epacanthaclisis banks, and Myrmeleontiformia is the monophyletic group.

The complete mitochondrial genome of Pycnarmon lactiferalis (Lepidoptera: Crambidae).

The complete mitochondrial genome of Pycnarmon lactiferalis (Walker, 1859) has been determined. The entire sequence is 15,219 bp in length which contains 13 protein-coding genes (PCGs), 22 transfer RNA genes, two ribosomal RNA genes, and an A + T-rich region. All PCGs start with the typical ATN codon except for COI with CGA. TAA is used for all the PCGs, while the COI, COII and ND5 possess incomplete termination codons T or TA. The secondary structure of 22 tRNAs have the typical clover-leaf pattern except for tRNASer(AGN) lacking the dihydrouridine (DHU) stem. The A + T-rich region, located between the srRNA and tRNAMet , do not contain the motif "ATAGA" that is conserved across other lepidopteran species mitogenomes. A phylogenetic relationship of Pyraloidea has been reconstructed based on 13 PCGs data using Bayesian inference (BI) method.