Journal: British Journal of Pharmacology

Article Title: A long‐acting FGF21 alleviates hepatic steatosis and inflammation in a mouse model of non‐alcoholic steatohepatitis partly through an FGF21‐adiponectin‐IL17A pathway

doi: 10.1111/bph.14383

Figure Lengend Snippet: The roles of IL17A in CD‐HFD‐induced NASH mice. (A) Flow cytometry analysis and quantification of IL17A expression on CD4+ T cells in the livers. (B) Relative mRNA levels of IL17A and Lcn2 in livers. (C) Western blot analysis of IL17A protein levels in the livers and quantitation of results of Western blot using Quantity One® (Bio‐Rad). (D) Serum IL17A was measured in CD‐HFD‐induced NASH mice with/without PsTag600‐FGF21. Data shown are means ± SD (n = 10 per group). *P < 0.05, significantly different from vehicle‐treated NASH mice.

Article Snippet: For rIL17A treatment, mice were injected i.p. with 25 μg·kg −1 recombinant mouse IL17A (Z03031, GenScript, Nanjing, China) twice a day for 15 days.

Techniques: Flow Cytometry, Expressing, Western Blot, Quantitation Assay

Journal: British Journal of Pharmacology

Article Title: A long‐acting FGF21 alleviates hepatic steatosis and inflammation in a mouse model of non‐alcoholic steatohepatitis partly through an FGF21‐adiponectin‐IL17A pathway

doi: 10.1111/bph.14383

Figure Lengend Snippet: The effect of recombinant IL17A on body weight, liver weight and glucose tolerance in NASH mice. (A) In a 15 day pharmacological study, change in body weight was monitored every day. NASH mice were injected i.p. with 3.7 mg·kg−1 PsTag600‐FGF21 every 3 days and 25 μg·kg−1 rIL17A twice a day. (B) Liver weight/body weight (%) of mice after 15 days of treatment. (C) For OGTT, blood glucose and the glucose AUC from 0 to 120 min were measured. Data shown are means ± SD (n = 10 per group). *P < 0.05, significantly different from vehicle‐treated NASH mice; # P < 0.05, significantly different from PsTag600‐FGF21‐treated NASH mice.

Article Snippet: For rIL17A treatment, mice were injected i.p. with 25 μg·kg −1 recombinant mouse IL17A (Z03031, GenScript, Nanjing, China) twice a day for 15 days.

Techniques: Recombinant, Injection

Journal: British Journal of Pharmacology

Article Title: A long‐acting FGF21 alleviates hepatic steatosis and inflammation in a mouse model of non‐alcoholic steatohepatitis partly through an FGF21‐adiponectin‐IL17A pathway

doi: 10.1111/bph.14383

Figure Lengend Snippet: PsTag600‐FGF21 indirectly suppressed Th17 cell differentiation and IL17A expression. (A) Naïve CD4+ T cells were treated with indicated concentrations of PsTag600‐FGF21 during differentiation to Th17 cells. The supernatant IL17A levels were assayed after differentiation. (B) Naïve CD4+ T cells were analysed for the expression of FGF21 receptor β‐klotho. Differentiated 3T3‐L1 adipocytes were used as a positive control. (C) Coculture of naïve CD4+ T cells and differentiated 3T3‐L1 adipocytes /AML12 cells/C2C12 myoblasts with/without PsTag600‐FGF21 (3.7 μg·mL−1) during differentiation to Th17 cells. Flow cytometry analysis and quantification of IL17A expression on CD4+ T cells after differentiation. (D) Co‐culture of differentiated 3T3‐L1 adipocytes and naïve CD4+ T cells were pretreated with GW9662 (10.0 μmol·L−1) for 1 h, followed by incubation with PsTag600‐FGF21 (3.7 μg·mL−1) during differentiation to Th17 cells. The supernatant IL17A levels were assayed after differentiation. Data shown are means ± SD (n = 6 per group). *P < 0.05, significantly different as indicated.

Article Snippet: For rIL17A treatment, mice were injected i.p. with 25 μg·kg −1 recombinant mouse IL17A (Z03031, GenScript, Nanjing, China) twice a day for 15 days.

Techniques: Cell Differentiation, Expressing, Positive Control, Flow Cytometry, Co-Culture Assay, Incubation

Journal: British Journal of Pharmacology

Article Title: A long‐acting FGF21 alleviates hepatic steatosis and inflammation in a mouse model of non‐alcoholic steatohepatitis partly through an FGF21‐adiponectin‐IL17A pathway

doi: 10.1111/bph.14383

Figure Lengend Snippet: PsTag600‐FGF21 suppressed Th17 cell differentiation via adiponectin (n = 5 per group). (A, B) Differentiated 3T3‐L1 adipocytes were incubated with indicated concentrations of PsTag600‐FGF21 for 24 h. Adiponectin concentration in the conditioned medium and relative adiponectin mRNA abundance were analysed. *P < 0.05, significantly different as indicated. (C) Naïve CD4+ T cells were treated with indicated concentrations of adiponectin during differentiation to Th17 cells. The levels of IL17A in the supernatants were assayed after differentiation. *P < 0.05, significantly different as indicated. (D) Naïve CD4+ T cells were analysed for the expression of the adiponectin receptors, Adipo1 and Adipo2. C2C12 muscle cells were used as a positive control. (E) Naïve CD4+ T cells were transfected with Adipo1 receptor siRNA or non‐targeting control siRNA. Relative abundance of Adipo1 receptor mRNA and protein levels were analysed. *P < 0.05, significantly different as indicated. (F) Naïve CD4+ T cells were transfected with Adipo1 receptor siRNA and were treated with adiponectin (5 μg·mL−1) during differentiation to Th17 cells. The levels of IL17A in the supernatants were assayed after differentiation. *P < 0.05, significantly different as indicated. (G) Differentiated 3T3‐L1 adipocytes were transfected with adiponectin siRNA or nontargeting control siRNA. Relative adiponectin mRNA abundance and protein levels were analysed. *P < 0.05, significantly different as indicated. (H) Differentiated 3T3‐L1 adipocytes were transfected with siRNA against adiponectin, or naïve CD4+ T cells were transfected with siRNA against Adipo1 receptors respectively. Differentiated 3T3‐L1 adipocytes and naïve CD4+ T cells were cocultured with PsTag600‐FGF21 (3.7 μg·mL−1) during differentiation to Th17 cells. The levels of IL17A in the supernatants were assayed after differentiation. *P < 0.05, significantly different as indicated. (I) Western blot analysis of IL17A protein levels in the livers was measured in PsTag600‐FGF21‐treated NASH mice with/without an anti‐adiponectin antibody. Quantitation of results of Western blot using Quantity One® (Bio‐Rad) *P < 0.05, significantly different from PsTag600‐FGF21‐treated NASH mice. (J) Serum IL17A in mice described in (I) (*P < 0.05 vs. PsTag600‐FGF21‐treated NASH mice). (K, L) Quantification of CD3 and F4/80 from Supporting Information Figure S7. Data shown are means ± SD. *P < 0.05, significantly different from PsTag600‐FGF21‐treated NASH mice.

Article Snippet: For rIL17A treatment, mice were injected i.p. with 25 μg·kg −1 recombinant mouse IL17A (Z03031, GenScript, Nanjing, China) twice a day for 15 days.

Techniques: Cell Differentiation, Incubation, Concentration Assay, Expressing, Positive Control, Transfection, Control, Western Blot, Quantitation Assay