Strictosamide alleviates acute lung injury via regulating T helper 17 cells, regulatory T cells, and gut microbiota.

BACKGROUND: Nauclea officinalis (Pierre ex Pit.) Merr. & Chun (Rubiaceae) is widely used to treat respiratory diseases in China. Strictosamide is its main active component and has significant anti-inflammatory activity. However, the effects and molecular mechanisms of strictosamide in the treatment of acute lung injury (ALI) remain largely unknown. PURPOSE: This study aimed to examine the regulatory effects of strictosamide on T helper 17 cells (Th17 cells)/Regulatory T cells (Treg cells) and gut microbiota in ALI-affected mice. MATERIALS AND METHODS: The ALI model was induced using lipopolysaccharide (LPS) intraperitoneal injection. Hematoxylin-eosin (H&E) staining, the number of inflammatory cells in broncho-alveolar lavage fluid (BALF), the Wet/Dry (W/D) ratio, and myeloperoxidase (MPO) activity were utilized as evaluation indices for the therapeutic efficacy of strictosamide on ALI. Flow cytometry (FCM), enzyme-linked immune sorbent assay (ELISA), quantitative reverse transcription polymerase chain reaction (qRT-PCR), and western blotting were used to determine the regulation of strictosamide on the Th17/Treg cells and the STAT3/STAT5 signaling pathway. The analysis of gut microbiota was conducted using 16S rDNA sequencing. The verification of the relationship between the gut microbiome and immune function was conducted using Spearman analysis. RESULTS: Strictosamide attenuated inflammation on ALI induced by LPS, which reduced the levels of Th17-related factors interleukin (IL)-6 and IL-17 and increased Treg-related factors IL-10 and transforming growth factor (TGF)-ß. In the spleens and whole blood, strictosamide reduced the proportion of Th17 cells and increased the proportion of Treg cells. Furthermore, strictosamide increased Forkhead/winged helix transcription factor 3 (Foxp3) and p-STAT5 protein expression while inhibiting Retinoid-related orphan nuclear receptors-γt (RORγt) and p-STAT3 expression. Moreover, strictosamide reshaped the diversity and structure of the gut microbiota, and influence the associations between immune parameters and gut microbiota in ALI mice. CONCLUSIONS: In summary, the results of the current investigation showed that strictosamide has a therapeutic impact on LPS-induced ALI. The mechanism of action of this effect may be associated with the modulation of Th17 and Treg cells differentiation via the SATA signaling pathway, as well as the impact of the gut microbiota.

Pterostilbene alleviated NAFLD via AMPK/mTOR signaling pathways and autophagy by promoting Nrf2.

BACKGROUND: NAFLD is a liver disease that is caused by liver damage or extreme lipid deposition but not alcohol. Nrf2 could mediate resistance to oxidative stress injury. Autophagy can degrade metabolic waste and accumulated toxic endogenous substances. Pterostilbene (PTE) is an active compound extracted from blueberry, and grape, that exhibits many biological effects, such as antiinflammation and antitumor. PURPOSE: This study provides a mechanism of PTE affecting on oxidative stress and autophagy in NAFLD mice. Tyloxapol, oil acid (OA) and palmitic acid (PA) were used to induce lipid accumulation in mice and HepG2 cells. METHODS: Western blotting, CRISPR/Cas 9 and other molecular biological approaches were applied to explore the mechanisms of PTE effected on NAFLD. RESULTS: PTE pretreatment effectively reduced the lipid accumulation in OA and PA induced HepG2 cells and tyloxapol induced mice, and significantly promoted the expression of nNrf2, PPAR-α and HO-1, and AMPK activity, but inhibited the expression of mTORC 1 and SREBP-1c. PTE activated phosphatidylinositide 3-kinase (PI3K) and proteins in the autophagy-related gene (ATG) family, and promoted the transformation of LC3Ⅰ to LC3Ⅱ which indicated the activation of autophagy, however, these effects were abolished after Nrf2 knockout. CONCLUSION: PTE effectively alleviated oxidative stress damage induced by excessive lipid accumulation in hepatocytes, thus promoting the metabolism and decomposition of fatty acids to improve NAFLD.

Uncovering the efficacy and mechanisms of Genkwa flos and bioactive ingredient genkwanin against L. monocytogenes infection.

ETHNOPHARMACOLOGICAL RELEVANCE: Genkwa flos (yuanhua in Chinese), the dried flower buds of the plant Daphne genkwa Siebold & Zucc., as a traditional herb widely used for the treatment of inflammation-related symptoms and diseases, with the efficacies of diuretic, phlegm-resolving and cough suppressant. AIM OF THE STUDY: Traditional Chinese Medicine (TCM) is presumed to be of immense potential against pathogens infection. Whereas, the potential efficacy and mechanisms of Genkwa flos against L. monocytogenes infection has not been extensively explored. The present study aimed to identify the bioactive ingredients of Genkwa flos against L. monocytogenes infection and to delineate the underlying mechanisms of action. MATERIALS AND METHODS: Bioinformatics approach at protein network level was employed to investigate the therapeutic mechanisms of Genkwa flos against L. monocytogenes infection. And hemolysis inhibition assay, cytoprotection test, western blotting, oligomerization assay and molecular docking analysis were applied to substantiate the multiple efficacies of Genkwa flos and the bioactive ingredient genkwanin. Histopathological analysis and biochemistry detection were conducted to evaluate the in vivo protective effect of genkwanin. RESULTS: Network pharmacology and experimental validation revealed that Traditional Chinese Medicine (TCM) Genkwa flos exhibited anti-L. monocytogenes potency and was found to inhibit the hemolytic activity of LLO. Bioactive ingredient genkwanin interfered with the pore-forming activity of LLO by engaging the active residues Tyr414, Tyr98, Asn473, Val100, Tyr440 and Val438, and thereby attenuated LLO-mediated cytotoxicity. Consistent with the bioinformatics prediction, exposed to genkwanin could upregulate the Nrf2 level and promote the translocation of Nrf2. In vivo, genkwanin oral administration (80 mg/kg) significantly protected against systemic L. monocytogenes infection, as evidenced by reduced myeloperoxidase (MPO) and malondialdehyde (MDA) levels, increased mice survival rate by 30% and decreased pathogen colonization. CONCLUSION: Our study demonstrated that Genkwa flos is a potential anti-L. monocytogenes TCM, highlighted the therapeutic potential of Genkwa flos active ingredient genkwanin by targeting the pore-forming cytolysin LLO and acting as a promising antioxidative candidate against L. monocytogenes infection.

Protective effects of morin on lipopolysaccharide/d-galactosamine-induced acute liver injury by inhibiting TLR4/NF-κB and activating Nrf2/HO-1 signaling pathways.

Morin, a bioactive flavonoid extracted from the bark of Moraceae plants and many medicinal herbs, has anti-inflammatory and antioxidative effects. In this research, we explored the protective effects of morin against lipopolysaccharide (LPS) and d-galactosamine (D-GalN) induced acute liver injury in mice. Mice were given an intraperitoneal injection of morin before LPS and D-GalN treatment and the HepG2 cells were only given morin to investigate its effects. The results showed that morin markedly inhibited the production of serum alanine transaminase (ALT), aspartate aminotransferase (AST), interleukin-6 (IL-6), tumor necrosis factor (TNF-α) and hepatic TNF-α, IL-6, and myeloperoxidase (MPO) induced by LPS/D-GalN. In order to evaluate morin effect in the future, we investigated the expression of nuclear factor E2 related factor 2 (Nrf2), nuclear factor-kappaB (NF-κB), toll like receptor 4 (TLR4) on liver injury. Taken together, these results suggested that morin could exert the anti-inflammatory and anti-oxidative effects against LPS/D-GalN-induced acute liver injury by activating Nrf2 signal pathways and inhibiting NF-κB activation.

[Diagnosis and treatment characteristics of head-wind sha in She medicine].

The diagnosis and treatment characteristics of head-wind sha in She medicine were analyzed and summarized. By visiting She-nationality villages and towns in Zhejiang province and Fujian province and interviewing hundreds of doctors of She medicine, the sha diagnosis, sha differentiation, experience and theory of treatment were arranged, and a comprehensive summary on theory and application of head-wind sha in She medicine such as pathogeny, name of disease, mechanism, diagnosis, differential diagnosis and treatment was made. It is believed that the methods of diagnosis and treatment in She medicine for head-wind sha could effectively enhance curative effect, safety and patients' quality of life, and the further research should be carried out.

[Comparative identification on Dai medicine "Pokou" and its processing product].

OBJECTIVE: To research identification methods of the Dai Medicine "Pokou" (the rhizome of Homalomena gigantea) and its processing product, and provide basis for identification of the drug in further research and application. METHODS: Macroscopic, microscopic observation and TLC and FTIR techniques were used to authenticate this raw medicine and its processing product. RESULTS: There were certain differences in the macroscopic features. The TLC result and infrared spectra of the samples had also obvious differences. The methods for identification of this raw medicine and its processing product were established, The detailed tissue and powder of this medicine were drawn. CONCLUSION: The results provided the basis for identification of the medicine and establishment of its quality standard.

[Effect of vitamin D supplementation in early life on the expression of interleukin-10 and intercellular adhesion molecule-1 in rat asthma model].

OBJECTIVE: To explore the effect and mechanism of vitamin D supplementation in early life on rat asthma model. METHOD: Thirty two sex-mature, female Wistar rats were randomly divided into a control group (n = 8), a low dose group (n = 8), a medium dose group (n = 8) and a high dose group (n = 8). From the seventh day of pregnancy on, the rats in each group were given different doses of vitamin D by intragastric administration, until the offspring rats were 21 days old. The rats in the control group were fed with DMSO-PBS. After the offsprings were weaned, 8 rats were randomly selected from each group. The number of male and female rats was equal. The rats were sensitized to ovalbumin (OVA) and challenged with aerosol OVA to establish the asthma model. The lung tissues were examined for pathologic changes after HE staining. ELISA was used to determine the concentrations of IL-10 in serum and BALF. Immunohistochemical staining methods were used to measure the expression of intercellular adhesion molecule-1 (ICAM-1) in lung tissues. RESULT: (1) Pathologic changes of lung tissues: compared with the control group, light microscope (LM) showed that eosinophil cells infiltration and the airway inflammation decreased in the low dose and medium dose groups, but increased in the high dose group. (2) The concentrations of IL-10 in serum and BALF: In serum, compared with the control group [(18.7 +/- 4.7) pg/ml], the concentrations of IL-10 in the low dose group [(30.2 +/- 2.8) pg/ml, P < 0.05] and the medium dose group [(51.5 +/- 6.6) pg/ml, P < 0.05] were significantly increased. And the IL-10 level of medium dose group was higher than that of the low dose group (P < 0.05). In BALF, compared with the control group [(59.1 +/- 14.4) pg/ml], the concentrations of IL-10 in the medium dose group [(90.0 +/- 14.3) pg/ml, P < 0.05] was significantly increased. There were no significant changes in the low dose group [(58.1 +/- 3.4) pg/ml, P > 0.05], whereas in the high dose group [(45.3 +/- 6.5) pg/ml, P < 0.05] the level significantly decreased. (2) The expression of ICAM-1 in lung tissues: compared with the control group, there were no significant changes in the low dose group (P > 0.05). The expression of ICAM-1 was significantly decreased in the medium dose group (P < 0.05). In the high dose group, the expression of ICAM-1 was significantly increased (P > 0.05). CONCLUSION: Adequate intervention with 1,25(OH)2D3 in the early life could alleviate the inflammation in the lung tissues, reduces eosinophil cell infiltration in rat asthma model. However, overdose might play a detrimental role. Its mechanism may be associated with the effect of 1,25(OH)2D3 on IL-10 secretion and the expression of ICAM-1.