Bofutsushosan (BTS; fangfengtongshengsan in Chinese) is a formula in traditional Japanese Kampo and Chinese medicine comprising 18 crude drugs and used to treat obesity and metabolic syndrome. In our previous study, BTS boiling water extract inhibited the uptake of fructose absorbed via glucose transporter 5 into cultured cells. In this study, the inhibitory effect of BTS extract on the absorption of fructose from the intestine was investigated in vivo. The extract of BTS was orally administered to rats at doses equivalent to 25-fold of the daily dose for humans. One minute after sample administration, fructose was orally administered and blood samples were collected from the jugular vein 0.5, 1, 1.5, 2, and 4 h after the administration of fructose. The absorption of fructose from the intestine was significantly reduced by treatment with BTS extract, and this in vivo study reproduced previous in vitro results. Subsequently, the blood samples were collected from the portal vein 30 min after the oral administration of fructose in mice. BTS extract significantly reduced fructose absorption in mice, and compared the effect of modified BTS samples by removing one to several crude drugs from BTS. We found that the dried rhizome of Rheum palmatum (RR) significantly contributed to the inhibitory effect of BTS on fructose absorption. We found sennoside A to be the active ingredient of RR for the inhibition of fructose absorption, and that its effect almost saturated at a dose of 3 mg/kg. These results support the action mechanisms of BTS when used for the treatment of obesity in clinics and drug stores. Graphical abstract
Physiologic responses mediated by hyaluronic acid (HAs) depend on their molecular weights and binding receptors, TLR4 and CD44. Low molecular weight HA (LMW-HA) induces inflammation, while high molecular weight HA (HMW-HA) works as an anti-inflammation factor. UVB-induced epidermal inflammation is usually initiated by endogenous molecules released from damaged cells upon skin tissue injury, called damaged-associated molecular patterns (DAMPs). Calprotectin, a typical DAMP, usually causes severe skin inflammation through activating TLR4 pathways. Since both LMW-HA and HMW-HA have inhibitory functions on TLR-mediated macrophage inflammation, HAs are assumed to suppress UVBinduced Calprotectin-mediated skin inflammations. In this study, ultra-LMW-HA, uLMW-HA (0.8 kDa) and HMW-HA (1200kDa), are utilized in the evaluation of UVB-irradiated keratinocyte inflammation. Calprotectin and proinflammatory cytokines secretion were demonstrated on UVB-irradiated keratinocytes. Results showed increased amounts of IL-6 and IL-8 in the medium after irradiation. Under HAs treatments on UVB-irradiated keratinocytes, IL-6 secretion showed a nearly 20% reduction(p<0.05), which indicates the anti-UVB-induced inflammatory effect of uLMW-HA and HMW-HA. Additionally, we treated the calprotectintreated keratinocytes with HAs since it was clear that UVB up-regulated Calprotectin secretion. Results showed suppressive effects of HAs on IL-6 secretion, and HMW-HA presented more substantial inhibitory effect compared to uLMW-HA. Furthermore, TLR4 downstream protein, TRAF6 expression was down-regulated in uLMW-HA-treated conditions, which supported uLMW-HA's blocking effects on Calprotectin/TLR4 signal. It is firmly believed that understanding the detailed molecular mechanisms of natural skin response to HAs will be helpful in guiding future HA-containing formulations designs.
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