Topical Ozone Accelerates Diabetic Wound Healing by Promoting Re-Epithelialization through the Activation of IGF1R-EGFR Signaling.

Ozonated oil increases the healing of chronic diabetic wounds, but the underlying mechanisms remain unclear. We investigated the effect of topical ozonated oil on wound healing in mice with diabetes with diet-induced obesity and further elucidated the role of EGFR and IGF1R signaling in diabetic wound healing. We found that topical ozonated oil accelerated wound healing; increased phosphorylation of IGF1R, EGFR, and VEGFR; and improved vascularization at the wound leading edge in mice with diabetes with diet-induced obesity. Exposure of normal epidermal keratinocytes to ozonated medium (20 µM for 2 hours daily) increased cell proliferation and migration distance by increasing phosphorylation of IGF1R and EGFR and downstream phosphoinositide 3-kinase, protein kinase B, and extracellular signal-regulated kinase. These findings shed light on the mechanism for topical ozone action in chronic wounds and support its potential therapeutic application.

[Inhibition of hepatitis C virus replication by mycophenolic acid in hepatocytes].

OBJECTIVE: It is well known that cyclosporine A (CsA), a widely used immunosuppressant for clinical organ transplantation, has the ability to inhibit HCV replication. In this study, the effects of several other immunosuppressants, including mycophenolic acid (MPA), rapamycin and FK-506, on HCV replication were examined in human hepatocytes. METHODS: HCV JFH-l-infected hepatocytes were treated with immunosuppressants or with control vehicles. The levels of viral RNA and the expression of HCV core protein were determined by quantitative real-time RT-PCR and Western Blot assay, respectively. RESULTS: MPA-treated cells showed significant decreases in both viral RNA and HCV Core protein expression compared with the control cells. Moreover, MPA treatments of hepatocytes before, during or after HCV infection could significantly inhibit viral replication. In contrast, rapamycin and FK-506 had little effect on HCV replication. Mechanism research disclosed that the inhibition of HCV replication by MPA was mainly due to its depletion of guanosine, a purine nucleoside crucial for synthesis of guanosine triphosphate (GTP), which is required for initiation of HCV RNA replication. The supplement of exogenous guanosine could reverse most of anti-HCV effect of MPA. CONCLUSION: These results indicate that MPA, through the depletion of guanosine, inhibits HCV JFH-1 replication in hepatocytes, suggesting that MPA may be beneficial for HCV-infected transplant recipients.

[Effect of TLR4 on the migration of asthmatic airway smooth muscle cells induced by airway epithelial cells].

OBJECTIVE: To explore the effect of Toll-like receptor 4 (TLR4) activation on the migration of asthmatic airway smooth muscle cell (ASMCs) induced by airway epithelial cells. METHODS: Primary ASMCs were cultured by the method of cell digestion. Cell culture supernatant of RTE cells were collected by TNF-alpha stimulation of epithelial cells. Detected the IL-8 and RANTES levels in the supernatant. The transmembrane migration of asthmatic ASMCs were detected by Modified Boyden chemotaxis chamber. The effect of TLR4 on the migration of asthmatic ASMCs induced by epithelial cells with TLR4 antibody drugs as a tool. RESULTS: The levels of IL-8 and RANTES in the supernatant of TNF-alpha groups were significantly increased, and that in the 20 ng/ml group was significantly higher than other groups (P < 0.01). The transmembrane migration of asthmatic ASMCs groups was increased than that of control group. The transmembrane migration of asthmatic ASMCs from asthma group and TNF-alpha + TLR4 antibody group was significantly decreased compared with that in TNF-alpha group (P < 0.01). The migration of asthma ASMCs from TNF-alpha + TLR4 antibody group was increased than that of asthma group (P < 0.05). CONCLUSION: TLR4 in the surface of asthmatic ASMCs may be activated by cytokines secreted by the airway epithelial cells and enhance the transmembrane migration of asthmatic ASMCs induced by airway epithelial cells so that it plays a role in airway remodeling of asthma.

[Effect of triptolide on airway smooth muscle proliferation and the expression of nuclear factor-kappa B, Bcl-2 in asthmatic rats].

OBJECTIVE: To explore the effect of triptolide on airway remodeling and the expression of nuclear factor-kappaB, Bcl-2 in asthmatic rats. METHODS: 40 rats were randomly divided into 5 groups (n = 8): (1) Control group; (2) Asthmatic 4 week group; (3) Asthmatic 6 week group; (4) Therapeutic 4 week group; (5) Therapeutic 6 week group. The airway resistance and eosinophilic inflammation of airway wall were observed. The airway wall thickness (WA/Pi), the bronchial smooth muscle thickness (smooth muscle area/Pi) and the number of bronchial smooth muscle nucleus (N/Pi) were measured by image analysis system. The expression of PCNA, nuclear factor-kappaB and Bcl-2 protein were determined by immunohistochemical staining and Western blot. The expression of Bcl-2 mRNA was determined by reverse transcription-polymerase chain reaction(RT-PCR). RESULTS: (1) The expression of NF-kappaB protein in asthmatic 4 week group and asthmatic 6 week group was significantly higher than that in control group, respectively (P < 0.01). The above-mentioned parameters of therapeutic 6 week group were significantly lower than those of asthmatic 4 week group, asthmatic 6 week group and therapeutic 4 week group, respectively (P < 0.01, P < 0.01 P < 0.05). (2) The expression of Bcl-2 protein and mRNA of asthmatic 4 week group and asthmatic 6 week group were significantly higher than those in control group respectively (P < 0.01). The expression of Bcl-2 protein of therapeutic 6 week group was significantly lower than those of asthmatic 4 week group, asthmatic 6 week group and therapeutic 4 week group respectively (P < 0.05, P < 0.01, P < 0.01), but the expression of Bcl-2 mRNA was significantly higher than the above-mentioned groups respectively (P < 0.01), the expression of Bcl-2 protein and mRNA of therapeutic 6 week group were higher than control group respectively (P < 0.05, P < 0.01). (3) The expression of PCNA protein of asthmatic 4 week group and asthmatic 6 week group were significantly higher than those of control group respectively (P < 0.01). (4) The WA/ Pi, the smooth muscle area/Pi and the N/Pi of asthmatic 4 week group and asthmatic 6 week group were significantly higher than those of control group, respectively (P < 0.01). The above-mentioned parameters of therapeutic 6 week group were significantly lower than those of asthmatic 4 week group, asthmatic 6 week group and therapeutic 4 week group, respectively (P < 0.01). (5) The airway resistance of asthmatic 4 week group and asthmatic 6 week group were significantly higher than those of the control group, respectively (P < 0.01). The above-mentioned parameters of therapeutic 6 week group were significantly lower than those of asthmatic 4 week group, asthmatic 6 week group and therapeutic 4 week group, respectively (P < 0.01, P < 0.01, P < 0.05). CONCLUSION: The proliferation of airway smooth muscle(ASM) is related with apoptosis of airway smooth muscle cells in asthma. NF-kappaB may be involved in the process. Triptolide may prevent apoptosis of ASMCs and decrease the proliferation of ASM by inhibiting the expression of NF-kappaB, Bcl-2.