Effect Of Taohong Siwu Decoction On Expression Of VEGF And Its Receptor In HDMEC Cells Injured By Ultraviolet Radiation

Purpose:This study through the simulated solar UV irradiation, from isolated cell level of observation of UVA to human skin dermal micro vascular endothelial cells(hdmec) damage and to the argument of activating blood circulation to dissipate blood stasis, on skin photoaging of syndrome pathogenesis, improve understanding of its mechanism, from the cellular and molecular level to clarify Taohong Siwu Decoction on human dermal micro vascular endothelial cells secrete vascular endothelial growth factor and its receptor expression of, elucidated in the repair of dermal microvascular endothelial cell injury, and against the skin photoaging mechanism.Material and method:This study by choosing one of dermal microvascular endothelial cells, with the method of homemade equipment, exposure to the sun’s ultraviolet rays, UVA light irradiation of model so as to establish a uv damage cells.Taohong Siwu Decoction preparing SD rats normal serum containing medicine serum and SD rats. Respectively establish HDMEC cell lines as follows: the blank control group(group A), UV model group(group B), UV + normal serum group(group C), UV + drug-containing serum group(group D), UV + retinoic acid group(group E) model of the five groups.(1) the blank control group(group A) : do not add any interventions, 24 h after normal training, collecting culture supernatant and cell, used for testing related indicators.(2) the model control group(group B) : will collapse HDMEC cell broth, PBS wash three times, add 2 MLPBS buffer, at UVA radiation meter, the dose of 10 j/cm2, from light source cells from 20 cm. Continue to develop 24 h after irradiation, germ-free collected 50 ml clear liquid, save the backup stored at 4 ℃; At the same time collecting cells, for related index detection.(3) the blank serum group(group C) : HDMEC cells after UVA irradiation method with group B(exposure), to join the culture medium containing 10% normal serum, cultivate 24 h, sterile and collect the 50 ml culture supernatant, stored at 4 ℃ in standby; At the same time collecting cells, for related index detection.(4) the drug-containing serum group(group D) : HDMEC cells after UVA irradiation method with group B(exposure), to join the culture medium containing 10% medicated serum, cultivate 24 h, sterile and collect the 50 ml culture supernatant, stored at 4 ℃ in standby; At the same time collecting cells, for related index detection.Retinoic acid(5) group(group E) : HDMEC cells after UVA irradiation method with group B(exposure), immediately join fang retinoic acid ethyl ester, clear liquid cultivation 24 h after collection and cells, for related index detection.Results:1. HDMEC cell morphological changesAdherent cell growth in blank control group, a polygon or short tail fusiform. Model control group, blank serum group and drug-containing serum group compared with blank control group, the cells become reduced, retraction, fuzzy boundaries, alignment, intercellular space increased obviously, and the drug-containing serum group cell damage degree is lower than the model control group and the blank serum group.2. VEGF content test resultsGroups of VEGF content in the cell culture supernatant on testing results show that the low content of VEGF in blank control group, Compared with blank control group, model control group and the blank serum VEGF levels were significantly higher in the group with statistical significance(P < 0.05), the drug-containing serum VEGF levels in the group also has a significantly increased compared with the blank control group, there is statistical significance(P < 0.05); Compared with model control group, medicated serum VEGF levels in the group improved significantly, there is statistical significance(P < 0.01); Compared with blank serum group, medicated serum VEGF levels in the group also has a significant boost, with significant difference(P < 0.01).3. VEGFR- 2 m RNA and protein expression level test resultsVEGFR- 2 groups of cells m RNA and protein expression level of the test results showed: the blank control group VEGFR- 2 m RNA and protein expression level is low, compared with blank control group, model control group and the blank serum group of VEGFR- 2 m RNA and protein expression level increased obviously and the differences are statistically significant(P < 0.05), serum medicated VEGFR- 2 m RNA and protein expression levels in the group compared with blank control group to compare rise sharply and there is statistical significance(P < 0.01); Group compared with model control group, serum medicated VEGFR- 2 m RNA and protein expression level increased obviously, there is significant difference(P < 0.01); Compared with blank serum group, serum medicated group of VEGFR-2 m RNA and protein expression level increased obviously, there is statistical significance difference(P < 0.01).Conclusion:1. From the point of view of cell morphology observation, Taohong Siwu Decoction drug-containing serum on after UVA irradiation damage HDMEC cells have a certain amount of repair effect.2. Taohong Siwu Decoction medicated serum could improve HDMEC cells secrete VEGF and VEGFR- 2 expression level,and promote the repair of dermal microvascular endothelial cells.