| Objective: Observing the influence of 5-Aminolevulinic acid photodynamic therapy associated with Salvia miltiorrhiza Bge. acting on Fibroblast derived from Hyperplastic scar and Vascular endothelial cell, and exploring its curative effect and mechanism.Methods: 1.Collecting the clinical samples: The samples of this experiment were taken from 12 patients’ scar tissue of The First People’s Hospital of Yunnan Province. It proves to be according with the clinical diagnosis by HE stain and microscope. 2.Separating and culturing FB and VEC derived from Hyperplastic scar, then picking out the tissue with a good condition during logarithmic phase to be used in experiment. 3.Grouping: The injection of the Salvia miltiorrhiza Bge. is confected into a few dose, including 1.5mg/ml, 3mg/ml, 6mg/ml, 12mg/ml and 24mg/ml.So is the laser, including 5J/cm2,10J/cm2,50J/cm2,100J/cm2,150J/cm2.And so is ALA, including 0.015mmol/L,0.15mmol/L,1.5mmol/L,15mmol/L,150mmol/L,1500mmol/L. Next,observe the form of the cells and measure the OD of every hole by ELISA with wavelength of 490 nm.Find the best effective dose beyond the 3 groups.Then mix these 3 factors up and make them act on the FB and VEC. Next,observe the form of the cells and measure the OD of every hole by ELISA with wavelength of 490 nm. 4.α-SMA,TGF-β1 and VEGF levels in supernatants were measured by ELISA. Theexpression of α-SMA protein content and TGF-β1 protein content were dectected by Western blotting. 5.With the way of statistic by software of SPSS19.0, the measurement data are showed by mean ± standard deviation, with P<0.05 having statistical significance.Results: The best concentration of the injections inhibiting Hyperplastic scar FB is 12mg/ml, inhibiting Hyperplastic scar VEC is 6mg/ml. The best light dose of laser acting on Hyperplastic scar FB and VEC is 100J/cm2 on average. The best concentration of the ALA acting on Hyperplastic scar FB and VEC is 1.5mmol/L. The OD in 490 nm of the ALA-PDT associated with Salvia miltiorrhiza Bge group on hypertrophic scar FB was 0.51±0.02, The OD in 490 nm of the ALA-PDT associated with Salvia miltiorrhiza Bge group on hypertrophic scar VEC was 0.62±0.02.Obviously, the group of ALA-PDT associated with Salvia miltiorrhiza Bge. shows better effect of inhibiting the Hyperplastic scar FB and VEC than the ones with only one method(P<0.01). All the groups were able to inhibit the secretion of α-SMA,TGF-β1 and VEGF. The concentration of α-SMA in the group of ALA-PDT associated with Salvia miltiorrhiza Bge was 19.60 ± 1.68 ng/ml, The concentration of TGF-β1 in the group of ALA-PDT associated with Salvia miltiorrhiza Bge was 13.47 ± 0.25 pg/ml, The concentration of VEGF in the group of ALA-PDT associated with Salvia miltiorrhiza Bge was 54.63 ± 1.42 pg/ml.The group of ALA-PDT associated with Salvia miltiorrhiza Bge. shows better effect of inhibiting the secretion of α-SMA,TGF-β1 and VEGF than the ones with only one method(P<0.01). All the groups were able to reduce the protein synthesis of α-SMA and TGF-β1, Under the action of ALA-PDT associated with Salvia miltiorrhiza Bge, the protein content of α-SMA was decreased by 81%, and the protein content of TGF-β1 was decreased by 64%, the group of ALA-PDT associated with Salvia miltiorrhiza Bge. shows better effect of reduceing of α-SMA and TGF-β1 than the ones with only one method.Conclusions:1、5-Aminolevulinic acid photodynamic therapy associated with Salvia miltiorrhiza Bge. acting can exert influence of inhibiting the morphology and proliferative activity of Hyperplastic Scar FB in vitro, whose effect is concentration-dependent and works on Hyperplastic Scar commendably. 2、5-Aminolevulinic acid photodynamic therapy associated with Salvia miltiorrhiza Bge. acting can exert influence of inhibiting the morphology and proliferative activity of Hyperplastic Scar VEC in vitro, whose effect is concentration-dependent and works on Hyperplastic Scar commendably. 3、ALA-PDT associated with Salvia miltiorrhiza Bge was able to inhibit the secretion of α-SMA,TGF-β1 and VEGF. 4、ALA-PDT associated with Salvia miltiorrhiza Bge was able to reduce the protein synthesis of α-SMA and TGF-β1. 5、ALA-PDT associated with Salvia miltiorrhiza Bge can produce better therapeutic effect on hypertrophic scar... |
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