雷帕霉素及去鐵敏對(duì)缺血缺氧創(chuàng)面愈合的影響
本文關(guān)鍵詞: 雷帕霉素 去鐵敏 雷帕霉素靶蛋白 缺氧誘導(dǎo)因子α 創(chuàng)面愈合 出處:《中國(guó)修復(fù)重建外科雜志》2017年06期 論文類型:期刊論文
【摘要】:目的探討雷帕霉素及去鐵敏對(duì)缺血缺氧創(chuàng)面愈合的影響及其作用機(jī)制。方法 SPF級(jí)雄性成年SD大鼠40只,體質(zhì)量(300±20)g,于背部制備缺血缺氧創(chuàng)面模型;將其隨機(jī)分為4組(n=10),分別為空白對(duì)照組(A組)、去鐵敏干預(yù)組(B組)、雷帕霉素干預(yù)組(C組)、去鐵敏+雷帕霉素共同干預(yù)組(D組)。模型制備后3、6、9 d,A、B、C、D組分別腹腔注射生理鹽水、去鐵敏(10 mg/kg)、雷帕霉素(3 mg/kg)、去鐵敏(10 mg/kg)+雷帕霉素(3 mg/kg)。模型制備后觀察創(chuàng)面愈合情況并記錄愈合時(shí)間;于創(chuàng)面完全愈合后第2天切取創(chuàng)面愈合組織,采用實(shí)時(shí)熒光定量PCR及Western blot檢測(cè)創(chuàng)面組織中雷帕霉素靶蛋白(mammalian target of rapamycin,m TOR)、缺氧誘導(dǎo)因子1α(hypoxia inducible factor 1α,HIF-1α)、VEGF m RNA及蛋白的表達(dá)。結(jié)果各組大鼠均成活至實(shí)驗(yàn)完成,創(chuàng)面均愈合;其中A、B、D組創(chuàng)面愈合時(shí)間均較C組明顯縮短(P0.05);A、B、D組間比較差異無統(tǒng)計(jì)學(xué)意義(P0.05)。實(shí)時(shí)熒光定量PCR檢測(cè)示,C、D組m TOR m RNA表達(dá)較A、B組明顯下調(diào)(P0.05),A、B組間比較差異有統(tǒng)計(jì)學(xué)意義(P0.05),C、D組間比較差異無統(tǒng)計(jì)學(xué)意義(P0.05)。B、D組HIF-1α、VEGF m RNA表達(dá)較A、C組明顯上調(diào),A組較C組表達(dá)上調(diào),比較差異有統(tǒng)計(jì)學(xué)意義(P0.05);B、D組間比較差異無統(tǒng)計(jì)學(xué)意義(P0.05)。Western blot檢測(cè)示,C、D組m TOR蛋白相對(duì)表達(dá)量較A、B組明顯下調(diào)(P0.05),C、D組間比較差異無統(tǒng)計(jì)學(xué)意義(P0.05)。A、B、C組HIF-1α蛋白相對(duì)表達(dá)量明顯高于D組(P0.05),A、B、C組間比較差異無統(tǒng)計(jì)學(xué)意義(P0.05)。B、C、D組VEGF蛋白相對(duì)表達(dá)量較A組明顯下調(diào),D組低于B、C組,C組低于B組,比較差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論去鐵敏可促進(jìn)大鼠缺血缺氧創(chuàng)面愈合,而雷帕霉素作用相反;可能與慢性缺血缺氧創(chuàng)面中存在m TOR與HIF-1信號(hào)調(diào)節(jié)通路有關(guān)。
[Abstract]:Objective to investigate the effect and mechanism of rapamycin and deferoxin on wound healing of ischemic and hypoxic wound. Methods 40 adult SD rats of SPF grade, with body weight of 300 鹵20 g, were used to establish the model of ischemic and hypoxic wound on the back. They were randomly divided into 4 groups: control group A, desferrin intervention group B, rapamycin intervention group C, destilmicin intervention group D, and normal saline injected intraperitoneally after 3 days, 6 days after the model was made, and 3 days after the model was made, normal saline was injected intraperitoneally in group A, group B, group B, group C, group C, group C, group C, group C, group C, group C, group C, group C, group C, group C, and group C, group C, group C, respectively. 10 mg / kg, rapamycin 3 mg / kg, rapamycin 10 mg / kg) rapamycin 3 mg / kg. The wound healing was observed and the healing time was recorded after the model was made, and the wound healing tissue was removed on the second day after the complete wound healing. Real-time fluorescence quantitative PCR and Western blot were used to detect the expression of rapamycin target protein mammalian target of rapamycin TORM, hypoxia inducible factor 1 偽 -hypoxia inducible factor 1 偽 -HIF-1 偽 -VEGFM-VEGF-M RNA and protein in wound tissue. There was no significant difference in wound healing time between group A and C (P 0.05). The expression of m TOR m RNA in group C was significantly lower than that in group A (P 0.05). There was a significant difference in the expression of m TOR m RNA between group A and C by real-time fluorescence quantitative PCR assay (RQFQ), which showed that the expression of m TOR m RNA in group C was significantly lower than that in group A (P 0.05) and the expression of m TOR m RNA in group C was significantly lower than that in group A (P 0.05). There was no significant difference between the two groups. The expression of HIF-1 偽 -VEGFM RNA in group P0.05 was significantly higher than that in group A and C, and the expression of VEGFM in group A was significantly higher than that in group C. There was no significant difference between the two groups. Western blot analysis showed that the relative expression of m TOR protein in group C was significantly lower than that in group A (P 0.05). There was no significant difference in the relative expression of HIF-1 偽 protein between group A and C (P 0.05). ABC group (P 0.05). The relative expression of HIF-1 偽 protein in group B was significantly lower than that in group A (P 0.05). The relative expression of HIF-1 偽 protein in group C was significantly lower than that in group A (P 0.05). There was no significant difference in the relative expression of HIF-1 偽 protein between two groups. The relative expression of VEGF protein in group D was significantly lower than that in group D (P 0.05), and the expression of VEGF protein in group C was lower than that in group B (P < 0.05), and the expression of VEGF protein in group C was lower than that in group B (P < 0.05), and the expression of VEGF protein in group D was significantly lower than that in group A (P 0.05). Conclusion Deferoxamine can promote the wound healing of ischemia and hypoxia in rats, but rapamycin has the opposite effect, which may be related to the existence of m TOR and HIF-1 signal regulation pathway in chronic ischemia and hypoxia wounds.
【作者單位】: 遵義醫(yī)學(xué)院附屬醫(yī)院燒傷整形外科;
【基金】:貴州省科學(xué)技術(shù)基金(黔科合J字[2011]2266號(hào))~~
【分類號(hào)】:R622
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