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.Monitoring and analysis of eye lens dose of interventional radiology workers
Qiuqiu PAN ; Jun DENG ; Mengxue LI ; Xiaohua MIAO ; Lingling ZHENG ; Yajuan FENG ; Senxing ZHENG
Chinese Journal of Radiological Health 2025;34(3):384-389
Objective To investigate the levels and influencing factors for eye lens dose of interventional radiology workers, and to provide a basis for reasonable and scientific radiation protection. Methods Thermoluminescent eye lens dosimeters were used to monitor the left and right eye lens doses of interventional radiology workers in real time during different surgical positions and varying eye protection conditions. The annual eye lens doses for the operators were estimated based on their yearly workload. The differences in eye lens doses under different conditions were analyzed and the influencing factors were identified. Results For individual interventional operations, the range of personal dose equivalent Hp(3) of the left eye of interventional radiology workers was ( < MDL ~ 418.33) μSv, the median (Q1, Q3) was 9.29 ( < MDL, 40.79) μSv, and the mean was 40.79 ± 70.36 μSv. The estimated annual eye lens doses were 4.05 mSv and 17.80 mSv based on the median and mean values of the eye lens dose of a single operation multiplied by average annual frequency of operations per person, respectively. The left eye lens dose was higher than the right eye lens dose of the same operator (Z = −4.24, P < 0.05), and the dose of the right eye lens was strongly positively correlated with that of the left eye lens. The left eye lens dose of the first surgeon was higher than that of the second surgeon in the same operation (Z = −3.10, P < 0.05). The eye lens dose was influenced by operator position (χ2 = 9.149, P = 0.002, OR = 8.343), eye protection (χ2 = 4.619, P = 0.032, OR = 4.352), and air kerma area product (χ2 = 8.032, P = 0.005, OR = 5.488). Conclusion According to the results of this study, a significant portion of interventional operators have eye lens doses that approach or exceed international occupational dose limits. It is recommended to pay attention to the operation frequency of the first operator and the air kerma area product of interventional operation, and strengthen radiation protection and dose monitoring for the eye lens of interventional radiology workers.