目的：本研究旨在观察黄连膏对全层皮肤缺损小鼠创面愈合的促进作用及其通过PI3K/AKT/eNos通路对血管生成的影响。方法：在45只C57BL/6J雄性小鼠背部制作直径为7.5 mm的全层皮肤切除模型，随机数字表法分为基质组、空白对照组和黄连膏组，基质组和黄连膏组创面分别涂抹基质和黄连膏治疗，空白对照组不涂抹药物。在造模后第0、3、7、10、14天时相观察创面皮肤的愈合情况；在造模后第3、7、14天各组分别处死5只小鼠取皮肤创面组织观察bFGF和PDGF的mRNA水平、CD-31细胞阳性数、AKT、VEGF-A、eNos的蛋白表达变化。对数据行单因素方差分析、重复方差分析和析因设计方差分析。结果：3组在0-7天创面愈合面积无明显差别，黄连膏组在第10、14天创面愈合率分别为（76±7）%和（93±5）%，与基质组（48±9）%和（68±11）%相比，P<0.01，差异有统计学意义；与空白对照组（46±7）%和（64±9）%相比，P<0.01，差异有统计学意义。结果显示黄连膏组伤后3、7 d创面组织中CD-31阳性细胞百分率分别为（16.3±3.2）%和(33.6±5.0)，明显高于基质组(12.5±4.6)%和(19.2±4.0)、空白对照组(8.4±2.4)%和(17.8±6.0)，(P<0.05或P<0.01)，差异有统计学意义。伤后3、7 d，黄连膏组小鼠创面组织中bFGF的mRAN水平分别为（1.75±0.22）和(2.98±0.35)，明显高于基质组（0.96±0.13）和(1.53±0.24)，空白对照组(0.78±0.24)%和(1.64±0.31)（P<0.01），差异有统计学意义；伤后14 d，基质组和空白对照 组小鼠创面组织中bFGF 的mRAN 水平分别为(1.88±0.38)和(2.03±0.21)达到最高，明显高于黄连膏组(1.43±0.42)，（P<0.05），差异有统计学意义。伤后3、7 d，黄连膏组小鼠创面组织中PDGF的mRAN水平（1.04±0.28）和 (1.82±0.25)，明显高于基质组（0.56±0.15）和(1.38±0.21)，（P<0.01或P<0.05），空白对照组(0.67±0.20)%和(1.45±0.26)（P<0.01或P<0.05）差异有统计学意义。黄连膏组伤后7 d，创面组织中P-AKTS308和P-AKTS437的蛋白分别 为（0.45±0.04）和(0.27±0.03)，明显高于基质组（0.23±0.06）和(0.16±0.04)、空白对照组（0.19±0.08）和(0.20±0.05), (P<0.05或P<0.01)，差异有统计学意义；VEGF-A和eNOS的蛋白含量（0.18±0.02）和(0.26±0.04)，也明显高于基质组（0.12±0.05）和(0.14±0.07)、空白对照组（0.13±0.06）和(0.17±0.03)，(P<0.01)，差异有统计学意义。结论：黄连膏能够提高全层皮肤缺损小鼠创面愈合率，并通过AKT/VEGF/eNos通路增加bFGF、PDGF、VEGF-A水平，促进模型小鼠创面的血管生成而促进创面愈合。

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 model was made in the back of the 45 male C57BL/6J mice respectively. That were subsequently randomly placed into 3 groups with Random number table method; i.e., vehicle, Huanglian ointment groups and the control group. In the Huanglian ointment group, topical Huanglian ointment was applied to the wound, in the vehicle group were treated with vehicle ointment, and in the control group were treated with nothing. Changes in the size of their wounds was monitored by taking pictures with a digital camera on days 0,3, 7, 10, and 14 after wound creation. The mice were sacrificed on the 3, 7, and 14 days after wound creation, and the tissue samples of the wounds were obtained for mRNA level of bFGF and PDGF、CD-31 cells and expression of AKT、VEGF-A、eNos were measured too. Results: Comparison of the sizes of the wounds among the groups showed that there was no significant difference on the 0,3 and 7 days, the most significant decreases were 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、7days, (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 mRAN of bFGF on the Huanglian ointment group were significantly higher than the vehicle and 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 than Huanglian ointment group, Huanglian ointment versus vehicle (1.43±0.42) VS(1.88±0.38), Huanglian ointment versus control (1.43±0.42) VS (2.03±0.21), P< 0.05. The mRAN of PDGF on the Huanglian ointment group were significantly higher 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 versus vehicle (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 significantly higher 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 ointment versus 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 eNOS on the Huanglian ointment group were significantly higher than the vehicle and control groups too, Huanglian ointment versus vehicle: (VEGF-A: (0.18±0.02) VS (0.26±0.04), P<0.01, eNOS: (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, eNOS: (0.12±0.05) VS ((0.17±0.03), (P<0.01)). Conclusions: Huanglian ointment promotes wound healing and enhance bFGF, PDGF and VEGF-A content by increasing the angiogenesis with the AKT/VEGF/eNos pathway.

上海市卫生和计划生育委员会卫中医药科研专项（2016JP006）：黄连膏促进全层皮肤切除小鼠创面愈合实验研究，负责人：奉典旭；上海市普陀区高层次人才科研创新项目（2014－A－24）：SnoN促进兔耳创面愈合及减轻兔耳增生性瘢痕的作用观察及机制研究，负责人：奉典旭；上海中医药大学创新团队（B－X－75）：中西医结合防治急性胰腺炎，负责人：奉典旭。