Zheng F, Cai Y E, Li L, Han D W
Department of Burns and Plastic Surgery, the First People's Hospital of Shangqiu, Shangqiu 476100, China.
Department of Burns, the First People's Hospital of Zhengzhou, Zhengzhou 450004, China.
Zhonghua Shao Shang Yu Chuang Mian Xiu Fu Za Zhi. 2023 Mar 20;39(3):256-263. doi: 10.3760/cma.j.cn501225-20220324-00087.
To investigate the effects and mechanism of astragalus polysaccharide (APS) on wound healing of deep partial-thickness burns in rats. The experimental study method was used. Fifty 7-week-old male Sprague-Dawley rats were divided into normal group, simple burn group, APS group, inhibitor group, and inhibitor+APS group according to the random number table, with 10 rats in each group. Except for normal group, rats in the other 4 groups were inflicted with a deep partial-thickness burn wound on the back. Rats in normal group and simple burn group were intraperitoneally injected with normal saline, and rats in the other three groups were injected with APS and/or integrin-linked kinase (ILK) inhibitor, respectively. After 7 days of injection, the wound healing rate of rats with burns in the four groups was calculated, and the serum content of interferon-γ, interleukin-2 (IL-2), and tumor necrosis factor α (TNF-α) in rats in 5 groups was determined by enzyme-linked immunosorbent assay (ELISA). The normal skin tissue of rats in normal group and wound tissue of rats with burns in the four groups were taken, the water content was determined and the water ratio was calculated, the content of interferon-γ, IL-2, and TNF-α was detected by ELISA, the mRNA expressions of epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), and ILK were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction, and the protein expressions of ILK, protein kinase B (Akt), phosphorylated Akt (p-Akt), glycogen synthetic kinase-3β (GSK-3β), and phosphorylated GSK-3β (p-GSK-3β) were detected by Western blotting. Data were statistically analyzed with one-way analysis of variance and least significant difference test. After 7 days of injection, the wound healing rate of rats in APS group was (67±5)%, which was significantly higher than (52±4)% in simple burn group and (59±5)% in inhibitor+APS group (with all the values <0.05). The wound healing rate of rats in inhibitor+APS group was significantly higher than (48±4)% in inhibitor group (<0.05). After 7 days of injection, compared with those in serum or normal skin tissue of rats in normal group, the serum content of interferon-γ, TNF-α, IL-2 and the water ratio of wound tissue of rats in simple burn group were significantly increased (<0.05); compared with those in APS group, the serum content of interferon-γ, TNF-α, IL-2 and the water ratio of wound tissue of rats in simple burn group and inhibitor+APS group were significantly increased (<0.05); compared with those in inhibitor group, the serum content of interferon-γ, TNF-α, IL-2 and the water ratio of wound tissue of rats in inhibitor+APS group were significantly decreased (<0.05). After 7 days of injection, compared with that in normal skin tissue of rats in normal group, the content of interferon-γ, TNF-α, and IL-2 in wound tissue of rats in simple burn group was significantly increased (<0.05); compared with that in APS group, the content of interferon-γ, TNF-α, and IL-2 in wound tissue of rats in simple burn group and inhibitor+APS group was significantly increased (<0.05); compared with that in inhibitor group, the content of interferon-γ, TNF-α, and IL-2 in wound tissue of rats in inhibitor+APS group was significantly decreased (<0.05). After 7 days of injection, compared with those in normal skin tissue of rats in normal group, the mRNA expressions of EGF, bFGF, ILK and protein expressions of ILK, p-Akt, p-GSK-3β in wound tissue of rats in simple burn group were significantly increased (<0.05); compared with those in APS group, the mRNA expressions of EGF, bFGF, ILK and protein expressions of ILK, p-Akt, p-GSK-3β in wound tissue of rats in simple burn group and inhibitor+APS group were significantly decreased (<0.05); compared with those in inhibitor group, the mRNA expressions of EGF, bFGF, ILK and protein expressions of ILK, p-Akt, p-GSK-3β in wound tissue of rats in inhibitor+APS group were significantly increased (<0.05). There were no statistically significant differences in the protein expressions of Akt and GSK-3β in normal skin tissue of rats in normal group and wound tissue of rats with burns in the four groups (>0.05). APS can alleviate systemic and local inflammation, alleviate tissue edema, and promote the expressions of healing factors in rats with deep partial-thickness burns, thus to promote the wound healing, possibly by activating ILK/Akt/GSK-3β signaling pathway.
探讨黄芪多糖(APS)对大鼠深Ⅱ度烧伤创面愈合的影响及其机制。采用实验研究方法。将50只7周龄雄性Sprague-Dawley大鼠按随机数字表分为正常组、单纯烧伤组、APS组、抑制剂组和抑制剂+APS组,每组10只。除正常组外,其他4组大鼠背部均造成深Ⅱ度烧伤创面。正常组和单纯烧伤组大鼠腹腔注射生理盐水,其他3组大鼠分别注射APS和/或整合素连接激酶(ILK)抑制剂。注射7天后,计算4组烧伤大鼠的创面愈合率,采用酶联免疫吸附测定(ELISA)法测定5组大鼠血清中干扰素-γ、白细胞介素-2(IL-2)和肿瘤坏死因子α(TNF-α)的含量。取正常组大鼠的正常皮肤组织和4组烧伤大鼠的创面组织,测定含水量并计算含水率,采用ELISA法检测干扰素-γ、IL-2和TNF-α的含量,采用实时荧光定量逆转录聚合酶链反应检测表皮生长因子(EGF)、碱性成纤维细胞生长因子(bFGF)和ILK的mRNA表达,采用蛋白质印迹法检测ILK、蛋白激酶B(Akt)、磷酸化Akt(p-Akt)、糖原合成激酶-3β(GSK-3β)和磷酸化GSK-3β(p-GSK-3β)的蛋白表达。数据采用单因素方差分析和最小显著差法进行统计学分析。注射7天后,APS组大鼠的创面愈合率为(67±5)%,显著高于单纯烧伤组的(52±4)%和抑制剂+APS组的(59±5)%(均P<0.05)。抑制剂+APS组大鼠的创面愈合率显著高于抑制剂组的(48±4)%(P<0.05)。注射7天后,与正常组大鼠血清或正常皮肤组织相比,单纯烧伤组大鼠血清中干扰素-γ、TNF-α、IL-2含量及创面组织含水率均显著升高(P<0.05);与APS组相比,单纯烧伤组和抑制剂+APS组大鼠血清中干扰素-γ、TNF-α、IL-2含量及创面组织含水率均显著升高(P<0.05);与抑制剂组相比,抑制剂+APS组大鼠血清中干扰素-γ、TNF-α、IL-2含量及创面组织含水率均显著降低(P<0.05)。注射7天后,与正常组大鼠正常皮肤组织相比,单纯烧伤组大鼠创面组织中干扰素-γ、TNF-α和IL-2含量显著升高(P<0.