1.Effect of hemoporfin-photodynamics on VEGF-A/MAPK/ERK pathway in vascular endothelial cells
Qichao LIU ; Yingjuan MEI ; Youquan LUO ; Mengxue ZHENG ; Rui YIN
Chinese Journal of Plastic Surgery 2020;36(4):385-391
Objective:To observe the molecular mechanism of Hemoporfin-mediated photodynamic therapy on vascular endothelial cells.Methods:Human umbilical vein endothelial cells (HUVEC) were cultured in vitro and divided into four groups: PDT group(Hemoporfin concentration: 5 μg/ml, light fluences: 4 J/cm 2), drug group (only Hemporfin: 5 μg/ml), light group(only irradiation by 4J/cm 2 light), and blank control (no drug, no light). The cell viability and proliferation were detected by cck-8 cytotoxicity test and Brdu testafter different treatments as mentioned above. Expression levels of VEGF-A/MAPK/ERK pathway related molecules in the cells before and after photodynamic treatment were detected by real-time quantitative PCR, Western blot and immunofluorescence staining. Results:Compared with the black control group, the cell viability[(0.45±0.08)vs(1.02±0.11), t=12.02, P<0.05] and cell proliferation level [(0.42±0.02)vs(1.00±0.01), t=31.20, P<0.05]were significantly decreased in PDT group.The mRNA expression levels, including Ras[(0.62±0.02)vs(1.05±0.03), t=10.35, P<0.05], c-Raf [(0.72±0.04)vs(1.00±0.05), t=7.35, P<0.05], Mek[(0.73±0.12)vs(1.15±0.04), t=7.74, P<0.05], Erk [(0.56±0.11)vs(1.02±0.03), t=5.56, P<0.05], VEGF-Α [(0.34±0.04)vs(1.02±0.07), t=7.59, P<0.05], and VEGFR2[(0.54±0.05)vs(1.00±0.03), t=5.34, P<0.05] were significantly decreased. The proteinphosphorylation level of c-Raf[(0.44±0.02)vs(1.02±0.05), t=46.7, P<0.05], Mek[(0.72±0.05)vs(1.05±0.04), t=5.35, P<0.05], Erk[(0.62±0.15)vs(1.03±0.03), t=8.58, P<0.05] and the proteinexpression level of VEGF-A[(0.64±0.03)vs(1.03±0.04), t=21.65, P<0.05] were significantly down-regulated in PDT group compared with the black control group. Compared with the blank control group, there were no significant differences expression between the drug group and the light group at cell activity, molecular proliferation level and molecular expressions. Conclusions:HMME-PDT inhibits the activity and proliferation of vascular endothelial cells by inhibiting the expression of the VEGF-A/MAPK/ERK pathway to achieve the purpose of inhibiting vascular hyperplasia and repair.
2.Effect of hemoporfin-photodynamics on VEGF-A/MAPK/ERK pathway in vascular endothelial cells
Qichao LIU ; Yingjuan MEI ; Youquan LUO ; Mengxue ZHENG ; Rui YIN
Chinese Journal of Plastic Surgery 2020;36(4):385-391
Objective:To observe the molecular mechanism of Hemoporfin-mediated photodynamic therapy on vascular endothelial cells.Methods:Human umbilical vein endothelial cells (HUVEC) were cultured in vitro and divided into four groups: PDT group(Hemoporfin concentration: 5 μg/ml, light fluences: 4 J/cm 2), drug group (only Hemporfin: 5 μg/ml), light group(only irradiation by 4J/cm 2 light), and blank control (no drug, no light). The cell viability and proliferation were detected by cck-8 cytotoxicity test and Brdu testafter different treatments as mentioned above. Expression levels of VEGF-A/MAPK/ERK pathway related molecules in the cells before and after photodynamic treatment were detected by real-time quantitative PCR, Western blot and immunofluorescence staining. Results:Compared with the black control group, the cell viability[(0.45±0.08)vs(1.02±0.11), t=12.02, P<0.05] and cell proliferation level [(0.42±0.02)vs(1.00±0.01), t=31.20, P<0.05]were significantly decreased in PDT group.The mRNA expression levels, including Ras[(0.62±0.02)vs(1.05±0.03), t=10.35, P<0.05], c-Raf [(0.72±0.04)vs(1.00±0.05), t=7.35, P<0.05], Mek[(0.73±0.12)vs(1.15±0.04), t=7.74, P<0.05], Erk [(0.56±0.11)vs(1.02±0.03), t=5.56, P<0.05], VEGF-Α [(0.34±0.04)vs(1.02±0.07), t=7.59, P<0.05], and VEGFR2[(0.54±0.05)vs(1.00±0.03), t=5.34, P<0.05] were significantly decreased. The proteinphosphorylation level of c-Raf[(0.44±0.02)vs(1.02±0.05), t=46.7, P<0.05], Mek[(0.72±0.05)vs(1.05±0.04), t=5.35, P<0.05], Erk[(0.62±0.15)vs(1.03±0.03), t=8.58, P<0.05] and the proteinexpression level of VEGF-A[(0.64±0.03)vs(1.03±0.04), t=21.65, P<0.05] were significantly down-regulated in PDT group compared with the black control group. Compared with the blank control group, there were no significant differences expression between the drug group and the light group at cell activity, molecular proliferation level and molecular expressions. Conclusions:HMME-PDT inhibits the activity and proliferation of vascular endothelial cells by inhibiting the expression of the VEGF-A/MAPK/ERK pathway to achieve the purpose of inhibiting vascular hyperplasia and repair.
3.Research on dynamic visual function difference
Jun CAI ; Dawei TIAN ; Po XU ; Yange ZHANG ; Qingjun ZHANG ; Xiaoyang YANG ; Yongsheng CHEN ; Lei SONE ; Mei LONG ; Yingjuan ZHENG ; Yuanyuan JIANG ; Shan CHEN ; Sujiang XIE ; Hongbo JIA ; Jie WANG
Military Medical Sciences 2014;(5):368-370
Objective To detect the dynamic visual acuity ( DVA) before and after vestibular habituation of subjects in order to optimize the DVA assessment criteria .Methods The vestibular function examination system was applied to the detection of static and dynamic visual function in 16 healthy subjects .Results When the speed of left or right swinging was fast enough , DVA before and after vestibular habituation was different .Conclusion Subjects with vestibular habituation can reduce their sensitivity to the vestibular system , the changes in DVA are better than before habituation , and the vestib-ular function adaptability training may have effect on DVA .