Abstract
Keishibukuryogan (KBG) is one of the traditional herbal formulations widely administered to patients with blood stagnation for improving blood circulation; currently, it is the most frequently prescribed medicine in Japan. KBG has been reported to improve conjunctional microcirculation. The aim of this study was to evaluate the role of KBG and paeoniflorin, a bioactive compound of KBG, in inhibiting the production of inflammatory cytokines using human dermal microvessel endothelial cells (HDMECs). The authors observed that lipopolysaccharide (LPS; 1 μg/mL) stimulated the secretion of proinflammatory cytokines in HDMECs. KBG treatment (10 mg/mL) significantly suppressed the mRNA levels of migration inhibitory factor (MIF), interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-α in LPS-stimulated cultured HDMECs. Similarly, paeoniflorin significantly suppressed the mRNA levels of these cytokines in LPS-stimulated cultured HDMECs. ELISA showed that KBG and paeoniflorin suppressed the production of MIF, IL-6, IL-8, and TNF-α in LPS-stimulated HDMECs. Moreover, KBG and paeoniflorin decreased the expression of cyclooxygenase-2 and inducible nitric oxide synthase (iNOS) in these cells. These results suggest that KBG may be useful for improving microvascular inflammation in patients with skin diseases.
Highlights
Keishibukuryogan (KBG, Gui-zhi-fu-ling-wan in Chinese) is one of the Kampo medicines, and has been widely administered to patients with blood stagnation for improving blood circulation
Louis); nylon membranes were from Schleicher & Schuell (Keene, NH); the anticyclooxygenase-2 (COX-2) polyclonal antibody was purchased from Cell Signaling Technology, Inc. (Boston); antiinducible NOS pAb was purchased from Enzo Life Sciences International Inc. (NY); the anti-βactin Ab was purchased from Santa Cruz Biotechnology Inc. (CA); Paeoniflorin was from LKT Laboratories, Inc.; migration inhibitory factor (MIF), IL-6, and IL-8 ELISA kits were obtained from R&D Systems (Minneapolis); and the Western blot detection system was obtained from Cell Signaling Technology (Beverly, MA)
Treatment of the cells with 1 μg/mL of LPS increased in mRNA levels of these cytokines from 6 h after stimulation, but these mRNA levels were decreased by KBG (Figure 1(c))
Summary
Keishibukuryogan (KBG, Gui-zhi-fu-ling-wan in Chinese) is one of the Kampo medicines, and has been widely administered to patients with blood stagnation for improving blood circulation. KBG has been used clinically to treat various diseases, including skin diseases. It was reported that KBG improves conjunctional microcirculation in patients with cerebrospinal vascular diseases [1], suggesting that it may have beneficial effects on hematological parameters such as blood viscosity and red blood cell deformability [2,3,4]. Matsumoto et al have explored a proteomic approach for the diagnosis of blood stasis in rheumatoid arthritis patients treated with KBG [5]. We recently reported that KBG is effective in patients with chronic pigmented purpura, a group of skin vascular disorders of unknown etiology [7]
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