AIM:To investigate the functions and potential mechanisms of obtusifolin in dry eye disease(DED)in a rat model.METHODS:A rat DED model was established via topical administration of benzalkonium chloride(BAC),followed ...AIM:To investigate the functions and potential mechanisms of obtusifolin in dry eye disease(DED)in a rat model.METHODS:A rat DED model was established via topical administration of benzalkonium chloride(BAC),followed by administration of obtusifolin.Conjunctival irritation score and tear production were measured to evaluate DED symptoms.Enzyme-linked immunosorbent assay(ELISA)was employed for determining inflammatory cytokine levels in rat conjunctiva.Periodic acid-Schiff staining and corneal fluorescein staining were implemented for assessing goblet cell numbers and corneal epithelial defects,respectively.Western blotting showed zonula occludens-1(ZO-1),matrix metalloproteinase-9(MMP-9),and mitogen-activated protein kinase(MAPK)/nuclear factor kappa B(NF-κB)signaling-related protein levels in the conjunctiva.RESULTS:Topical application of obtusifolin alleviated conjunctival irritation and enhanced tear production in BAC-induced DED rats.Obtusifolin attenuated conjunctival inflammatory response and goblet cell loss as well as corneal epithelial barrier disruption in DED rats.Obtusifolin suppressed extracellular signal-regulated kinase(ERK),p38,and NF-κB phosphorylation in the conjunctiva of DED rats.CONCLUSION:Obtusifolin ameliorates DED in rats possibly by alleviating inflammation via the inactivation of MAPK/NF-κB signaling.展开更多
AIM: To explore the effect of Obtusifolin on retinal pigment epithelial cell growth under hypoxia.METHODS: In vitro chemical hypoxia model of ARPE-19 cells was established using cobalt chloride(CoCl2). Cell viability ...AIM: To explore the effect of Obtusifolin on retinal pigment epithelial cell growth under hypoxia.METHODS: In vitro chemical hypoxia model of ARPE-19 cells was established using cobalt chloride(CoCl2). Cell viability was tested by cell counting kit-8(CCK-8) assay. Western blot and real-time quantitative polymerase chain reaction were applied to detect proteins and mRNAs respectively. Flow cytometry was used to examine the cell cycle. Secretion of vascular endothelial growth factor(VEGF) was tested by using enzyme linked immunosorbent assay(ELISA).RESULTS: Under the chemical hypoxia model established by CoCl2, hypoxia inducible factor-1α(HIF-1α) mRNA and protein levels was up-regulated. Cell viability was increased and the proportion of S phase was higher. Obtusifolin could reduce cell viability under hypoxic conditions and arrest cells in G1 phase. Obtusifolin reduced the expression of Cyclin D1 and proliferating cell nuclear antigen(PCNA) in the hypoxic environment and increased the expression of p53 and p21. The levels of VEGF, VEGFR2 and eNOS proteins and mRNA were significantly increased under hypoxia while Obtusifolin inhibited the increasing.CONCLUSION: Obtusifolin can inhibit cell growth under hypoxic conditions and down-regulate HIF-1/VEGF/eNOS secretions in ARPE-19 cells.展开更多
基金Supported by Hubei Provincial Hospital of Traditional Chinese Medicine In-hospital Project(No.2021YJKT-20)Hubei Provincial Administration of Traditional Chinese Medicine,Bureau-level Project(No.ZY2023F008)Hubei Science and Technology Department,General Project(No.2025AFB786).
文摘AIM:To investigate the functions and potential mechanisms of obtusifolin in dry eye disease(DED)in a rat model.METHODS:A rat DED model was established via topical administration of benzalkonium chloride(BAC),followed by administration of obtusifolin.Conjunctival irritation score and tear production were measured to evaluate DED symptoms.Enzyme-linked immunosorbent assay(ELISA)was employed for determining inflammatory cytokine levels in rat conjunctiva.Periodic acid-Schiff staining and corneal fluorescein staining were implemented for assessing goblet cell numbers and corneal epithelial defects,respectively.Western blotting showed zonula occludens-1(ZO-1),matrix metalloproteinase-9(MMP-9),and mitogen-activated protein kinase(MAPK)/nuclear factor kappa B(NF-κB)signaling-related protein levels in the conjunctiva.RESULTS:Topical application of obtusifolin alleviated conjunctival irritation and enhanced tear production in BAC-induced DED rats.Obtusifolin attenuated conjunctival inflammatory response and goblet cell loss as well as corneal epithelial barrier disruption in DED rats.Obtusifolin suppressed extracellular signal-regulated kinase(ERK),p38,and NF-κB phosphorylation in the conjunctiva of DED rats.CONCLUSION:Obtusifolin ameliorates DED in rats possibly by alleviating inflammation via the inactivation of MAPK/NF-κB signaling.
文摘AIM: To explore the effect of Obtusifolin on retinal pigment epithelial cell growth under hypoxia.METHODS: In vitro chemical hypoxia model of ARPE-19 cells was established using cobalt chloride(CoCl2). Cell viability was tested by cell counting kit-8(CCK-8) assay. Western blot and real-time quantitative polymerase chain reaction were applied to detect proteins and mRNAs respectively. Flow cytometry was used to examine the cell cycle. Secretion of vascular endothelial growth factor(VEGF) was tested by using enzyme linked immunosorbent assay(ELISA).RESULTS: Under the chemical hypoxia model established by CoCl2, hypoxia inducible factor-1α(HIF-1α) mRNA and protein levels was up-regulated. Cell viability was increased and the proportion of S phase was higher. Obtusifolin could reduce cell viability under hypoxic conditions and arrest cells in G1 phase. Obtusifolin reduced the expression of Cyclin D1 and proliferating cell nuclear antigen(PCNA) in the hypoxic environment and increased the expression of p53 and p21. The levels of VEGF, VEGFR2 and eNOS proteins and mRNA were significantly increased under hypoxia while Obtusifolin inhibited the increasing.CONCLUSION: Obtusifolin can inhibit cell growth under hypoxic conditions and down-regulate HIF-1/VEGF/eNOS secretions in ARPE-19 cells.
