Figure 6.

Sch B reduces inflammatory responses in db/db mice by inhibiting MyD88. A) Co‐immunoprecipitation showing formation of MD2/TLR4, TLR4/MyD88, and TLR4/TRIF complexes in heart tissues of db/db mice. TLR4 was immunoprecipitated (IP) and levels of MD2, TRIF, and MyD88 were detected by immunoblotting (IB). Densitometric quantification is shown on right. B) Western blot analysis of pathway activation downstream of MyD88 in heart tissues. Phosphorylated TAK1, TBK1, and IRF3 were detected. Total proteins were used as control. Densitometric quantification shown on right. C) Analysis of MAPK and NF‐κB activation in heart tissues of db/db mice. Phosphorylated ERK, JNK, and p38 were detected. Total proteins were used as control. IκB‐α was used to measure NF‐κB activity, with GAPDH as the loading control. Right panel shows densitometric quantification. D) mRNA levels of Tnf, Il6, and Ifng in heart tissues of mice. Transcripts were normalized to Actb. E) Protein levels of IL‐6 and TNF‐α in serum of db/db mice were determined by enzyme‐linked immunoassay (ELISA). Data in all panels are shown as mean ± SEM (n = 6 per group; *p < 0.05, **p < 0.01, ***p < 0.001 compared to db/m; #p < 0.05, ##p < 0.01, ns = not significant compared to db/db by one‐way ANOVA followed by Bonferroni's multiple comparisons test).