Effects of Huanglian Ointment promoting Angiogenesis by PI3K/AKT/eNos pathway in Full-thickness Skin Defect Mice / 世界科学技术-中医药现代化

ABSTRACT

Objective: to observe the effects of Huanglian ointment promote wound healing and angiogenesis by the AKT/VEGF/eNos pathway in full-thickness skin defect mice. Methods: 7.5 mm diameter full-thickness skin excision modelwas made in the back of the 45 male C57 BL/6 J mice respectively. That were subsequently randomly placed into 3 groupswith Random number table method; i.e., vehicle, Huanglian ointment groups and the control group. In the Huanglianointment group, topical Huanglian ointment was applied to the wound, in the vehicle group were treated with vehicleointment, and in the control group were treated with nothing. Changes in the size of their wounds was monitored by takingpictures with a digital camera on days 0, 3, 7, 10, and 14 after wound creation. The mice were sacrificed on the 3, 7, and14 days after wound creation, and the tissue samples of the wounds were obtained for m RNA level of b FGF and PDGF、CD-31 cells and expression of AKT、VEGF-A、eNos were measured too. Results: Comparison of the sizes of the woundsamong the groups showed that there was no significant difference on the 0, 3 and 7 days, the most significant decreaseswere found in experimental Huanglian ointment group on the day10 (Huanglian ointment versus vehicle (76±7) ％ VS (48±9) ％, huanglian ointment versus control (76±7) ％ VS (46±7) ％, P＜0.01), and day14 (Huanglian ointment versus vehicle (93±5) ％ VS (68 ±11) ％, huanglian ointment versus control (93 ±5) ％ VS (64±9) ％, P＜0.01) . The percentage of CD-31 cells on the Huanglian ointment group were significantly higher than that of the vehicle and control groups on the 3、7 days, (Huanglian ointment versus vehicle day3 (16.3±3.2) ％ VS (12.5±4.6) ％, P＜0.05;day7 (33.6±5.0) ％ VS (19.2±4.0) ％, P＜0.01; (Huanglian ointment versus control day3 (16.3±3.2) ％ VS (8.4±2.4) ％, P＜0.05;day7 (33.6±5.0) ％ VS (17.8±6.0) ％, P＜ 0.01. The m RAN of b FGF on the Huanglian ointment group were significantly higher than the vehicleand control groups on the 3 and 7 days, (day3 Huanglian ointment versus vehicle (1.75±0.22) VS (0.96±0.13), day7 (2.98±0.35) VS (1.53±0.24), P＜0.01) ); (day3 Huanglian ointment versus control (1.75±0.22) VS (0.78±0.24), and day7 (2.98 ± 0.35) VS (1.64 ± 0.31), P＜0.01) . But on 14 day, the vehicle and control groups were significantly higher thanHuanglian ointment group, Huanglian ointment versus vehicle (1.43±0.42) VS (1.88±0.38), Huanglian ointment versuscontrol (1.43±0.42) VS (2.03±0.21), P＜ 0.05. The m RAN of PDGF on the Huanglian ointment group were significantlyhigher than the vehicle and control groups on the 3、7 days, (day3 Huanglian ointment versus vehicle (1.04±0.28) VS (0.56±0.15), Huanglian ointment versus control (1.04±0.28) VS (0.67±0.20) (P＜0.01); day7 Huanglian ointment versusvehicle (1.82±0.25) VS (1.38±0.21), Huanglian ointment versus control (1.82±0.25) VS (1.45±0.26) (P＜0.05) . On the 7 day, the protein of P-AKTS308 and P-AKTS437 in wound tissue on the Huanglian ointment group were significantlyhigher than the vehicle and control groups, (P-AKTS308 Huanglian ointment versus vehicle (0.45±0.04) VS (0.23±0.06), Huanglian ointment versus control (0.45 ± 0.04) VS (0.19 ± 0.08), (P＜0.05); (P-AKTS437 Huanglian ointmentversus vehicle (0.27±0.03) VS (0.16±0.04); Huanglian ointment versus control (0.27±0.03) VS (0.20±0.05), (P＜0.01) .the protein of VEGF-A and e NOS on the Huanglian ointment group were significantly higher than the vehicle and controlgroups too, Huanglian ointment versus vehicle (VEGF-A (0.18±0.02) VS (0.26±0.04), P＜0.01, e NOS (0.12±0.05) VS (0.14±0.07, P＜0.01) ); Huanglian ointment versus control (VEGF-A (0.18±0.02) VS (0.13±0.06), P＜0.01, e NOS (0.12±0.05) VS ((0.17±0.03), (P＜0.01) ) . Conclusions: Huanglian ointment promotes wound healing and enhance b FGF, PDGFand VEGF-A content by increasing the angiogenesis with the AKT/VEGF/eNos pathway.