1. miR‐181s can induce immunoparalysis AGroups of mice were subjected to CLP, followed 2 days later by injection with agomir‐181d or agomir‐NC via tail vein at the dose of 10 nmol per mouse once a day for three consecutive days.
miR‐181s can induce immunoparalysis AGroups of mice were subjected to CLP, followed 2 days later by injection with agomir‐181d or agomir‐NC via tail vein at the dose of 10 nmol per mouse once a day for three consecutive days. The mortality of mice after CLP 48 h was controlled at 20% (punctured once). Data shown are from one experiment (n = 20 mice per group), representative of a total of three independent experiments. *P =  (log‐rank test). BBacterial loads in the blood (left panel) and spleen (right panel) of CLP mice after injection with agomir‐181d or agomir‐NC via tail vein for 2 days. Data shown are from one experiment (n = 6–10 mice per group), representative of a total of three independent experiments. *P = 0.001, **P =  (Student's t‐test). CAfter injection with agomir‐181d or agomir‐NC via tail vein for 2 days, the count of CD4+ or CD8+ T cells in the spleen (left panel) and lymph nodes (right panel) of CLP mice was analyzed by flow cytometry, as described in the method section. Sham‐operated mice group was used as a control. Data are expressed as mean ± SD, n = 6–10 mice per group and are representative of three experiments. *P = 0.0001, **P = 0.0012, ***P =  (one‐way ANOVA). N.S., not statistically significant. DAntagomir‐181 reversed LPS‐induced endotoxin tolerance. Monocytes from 5 healthy volunteers were isolated, pre‐treated with 800 pmol antagomir‐181 or matched antagomir control for 24 h, further treated with 1 μg/ml LPS for 16 h, and then re‐stimulated with 1 μg/ml LPS for 2 h. TNF‐α mRNA levels were determined by Q‐PCR. Data are expressed as mean ± SD from two independent experiments. *P =  (one‐way ANOVA); N.S., not statistically significant. E, FInverse correlation between the expressions of miR‐181s (E) and TNF‐α mRNA (F) in monocytes from sepsis patients. Monocytes of 25 patients with severe sepsis and 22 healthy blood donors were isolated by adherence to plastic substrates. The culture plates were incubated for 2 h in a humidified 37°C, 5% CO2 incubator. After incubation, the media containing non‐adherent cells were removed by aspiration. Total RNA of the adherent cells was extracted using RNeasy kit (Invitrogen, USA). miR‐181s and TNF‐α mRNA levels were determined by Q‐PCR. Data are expressed as mean ± SD. *P = 0.0054, **P = 0.0034, ***P = 0.0309, ****P =  (Student's t‐test).
Cao Dan et al. EMBO Mol Med. 2015;7:
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