[Phenylhexyl isothiocyanate induces gene p15 demethylation by down-regulating DNA methyltransferases in Molt-4 cells].

This study is to investigate the effect of phenylhexyl isothiocyanate (PHI), which has been proved to be a novel histone deacetylase inhibitor (HDACi) recently, on gene p15 de novo expression in acute leukemia cell line Molt-4, and to further study its potential mechanism. Modified methylation specific PCR (MSP) was used to screen p15-M and p15-U mRNA. DNA methyltransferasel (DNMT1), 3A (DNMT3A), 3B (DNMT3B) and p15 mRNA were measured by RT-PCR. P15 protein was detected by Western blotting. Hypermethylation of gene p15 was reversed and activation transcription of gene p15 in Molt-4 was de novo after 5 days exposure to PHI in a concentration dependent manner. DNMT1 and DNMT3B were inhibited by exposure to PHI for 5 days (P < 0.05). Alteration of DNMT3A was not significant. It is showed that PHI could reverse hypermethylation of gene p15 and transcriptional activation of gene p15 is de novo by PHI. It may result from down-regulating DNA methyltransferases, DNMT1 and DNMT3B, or up-regulating the histone acetylation that allows chromatin unfolding and the accessibility of regulators for transcriptional activation in the p15 promoter.

[Effects of acupoint-injection of Lentinan on the immunologic funtion in rabbits with spleen-qi deficiency].

UNLABELLED: To observe the effect of Spleen-Meridian-acupoint injection of Lentinan on the immunologic function in spleen-deficiency rabbits. METHODS: A total of 54 Newzealand rabbits were randomly divided into normal (n = 10), model (n = 8), intramuscular injection (n = 10), Sanyinjiao (SP6, n = 10), Diji (SP8, n = 8) and Xuehai (SP 12, n = 8) groups. Spleen-qi deficiency model was established by intragastric administration of 100% crude Radix et Rhizoma Rhei decoction (15 mL/kg/day x 10 d), and then Lentinan (LNT, 0.025 mg/kg/2 day x 5) was injected into the aforementioned acupoints of the Spleen Meridian. The erythrocyte immunologic function (RBC-C3 bR, RBC-IC), hemolysin (lgM) and changes of physical signs of the rabbits were observed. RESULTS: In comparison with the control group, the rabbits' body weight, rectal temperature, RBC-C3 bR% and serum IgM level were decreased significantly in model group (P < 0.05); while in comparison with the model group, the body weight in SP 8 group,retal temperature in SP 9 and SP 8 groups, RBC-C3 bR% in SP 9 and SP 12 groups, and serum IgM levels in SP 9, SP 8 and SP 12 groups increased considerably (P < 0.05, P < 0.01). Comparison among the 4 treatment groups showed that the effect of SP 12 was superior to that of intramuscular injection group in upregulaing RBC-C3 bR%, and the effects of SP 9, SP 8 and SP 12 groups were significantly superior to those of intramuscular injection group in upregulating serum IgM level (P < 0.05). CONCLUSION: The Spleen-Meridian-acupoint injection of LNT is superior to that of intramuscular injection of LNT in improving the spleen-qi deficiency rabbits' symptoms and immunologic function.

Diosmetin-7-O-ß-D-glucopyranoside from Pogostemonis Herba alleviated SARS-CoV-2-induced pneumonia by reshaping macrophage polarization and limiting viral replication.

ETHNOPHARMACOLOGICAL RELEVANCE: Viral pneumonia is the leading cause of death after SARS-CoV-2 infection. Despite effective at early stage, long-term treatment with glucocorticoids can lead to a variety of adverse effects and limited benefits. The Chinese traditional herb Pogostemonis Herba is the aerial part of Pogostemon Cablin (Blanco) Benth., which has potent antiviral, antibacterial, anti-inflammatory, and anticancer effects. It was used widely for treating various throat and respiratory diseases, including COVID-19, viral infection, cough, allergic asthma, acute lung injury and lung cancer. AIM OF THE STUDY: To investigate the antiviral and anti-inflammatory effects of chemical compounds from Pogostemonis Herba in SARS-CoV-2-infected hACE2-overexpressing mouse macrophage RAW264.7 cells and hACE2 transgenic mice. MATERIALS AND METHODS: The hACE2-overexpressing RAW264.7 cells were exposed with SARS-CoV-2. The cell viability was detected by CCK8 assay and cell apoptotic rate was by flow cytometric assay. The expressions of macrophage M1 phenotype markers (TNF-α and IL-6) and M2 markers (IL-10 and Arg-1) as well as the viral loads were detected by qPCR. The mice were inoculated intranasally with SARS-CoV-2 omicron variant to induce viral pneumonia. The levels of macrophages, neutrophils, and T cells in the lung tissues of infected mice were analyzed by full spectrum flow cytometry. The expressions of key proteins were detected by Western blot assay. RESULTS: Diosmetin-7-O-ß-D-glucopyranoside (DG) presented the strongest anti-SARS-CoV-2 activity. Intervention with DG at the concentrations of 0.625-2.5 µM not only reduced the viral replication, cell apoptosis, and the productions of inflammatory cytokines (IL-6 and TNF-α) in SARS-CoV-2-infected RAW264.7 cells, but also reversed macrophage polarity from M1 to M2 phenotype. Furthermore, treatment with DG (25-100 mg/kg) alleviated acute lung injury, and reduced macrophage infiltration in SARS-COV-2-infected mice. Mechanistically, DG inhibited SARS-COV-2 gene expression and HK3 translation via targeting YTHDF1, resulting in the inactivation of glycolysis-mediated NF-κB pathway. CONCLUSIONS: DG exerted the potent antiviral and anti-inflammatory activities. It reduced pneumonia in SARS-COV-2-infected mice via inhibiting the viral replication and accelerating M2 macrophage polarization via targeting YTHDF1, indicating its potential for COVID-19 treatment.