中国病理生理杂志2023，Vol.39Issue(12)：2223-2233,11.DOI:10.3969/j.issn.1000-4718.2023.12.013
硫化氢对糖尿病大鼠皮肤创面自噬和血管形成的影响
Effects of hydrogen sulfide on autophagy and angiogenesis of skin wound in diabetic rats
摘要
Abstract
AIM:To explore the effect and mechanism of hydrogen sulfide(H2S)on autophagy and angiogene-sis in skin wound of diabetic rats.METHODS:Among 36 healthy 8-week-old male Sprague-Dawley rats,12 rats were se-lected as control group,and the remaining rats were intraperitoneally injected with streptozotocin(STZ)to induce diabetic model and were randomly divided into diabetes mellitus(DM)group and NaHS(H2S donor)intervention(DM+NaHS)group,with 12 rats in each group.A skin trauma model was established by excising the skin of the back of rats in each group.The rats in DM+NaHS group were intraperitoneally injected with NaHS(56 μmol/kg),and the rats in control and DM groups were daily received the same volume of normal saline for 21 consecutive days.The healing of skin wound was measured on days 0,7,14 and 21 after operation.On the 21st day after surgery,the content of H2S in skin tissues was de-tected by C-7Az fluorescent probe,and the morphological changes and angiogenesis of wound tissues were observed by HE staining.The expression of CD31 was detected by immunofluorescence staining,and endothelial autophagy was detected by double staining of CD31 and beclin-1.The protein levels of cystathionine γ-lyase(CSE),CD31,microtubule-associated protein 1 light chain 3(LC3),beclin-1,P62,Bcl-2,Bax,phosphatidylinositol 3-kinase(PI3K),protein kinase B(PKB/Akt)and mammalian target of rapamycin(mTOR)in wound tissues were determined by Western blot.Caspase-3 and propidium iodide(PI)staining was used to detect cell apoptosis,and apoptosis of vascular endothelial cells was deter-mined with CD31 and TUNEL double immunofluorescence staining.RESULTS:Compared with DM group,the wound healing rate,H2S content and CSE protein expression were significantly increased in DM+NaHS group(P＜0.01),but still lower than those in control group(P＜0.01).HE staining showed that the wound surface in DM group was thin and wide,with few capillary,while that in DM+NaHS group was thicker with lots of capillary and wound width was reduced.Com-pared with DM group,CD31 expression was markedly increased(P＜0.01),the fluorescence intensity of caspase-3 and PI was significantly decreased(P＜0.01),and CD31+/beclin-1+ as well as CD31+/TUNEL+ cells were decreased(P＜0.01)in DM+NaHS group.Western blot analysis showed that compared with DM group,the levels of beclin-1,Bax and LC3-Ⅱ/LC3-Ⅰ were significantly decreased(P＜0.01),while the levels of P62 and Bcl-2,as well as ratios of p-PI3K/PI3K,p-Akt/Akt and p-mTOR/mTOR were significantly increased(P＜0.01)in DM+NaHS group.CONCLUSION:H2S can promote skin wound healing,which may be related to activation of PI3K/Akt/mTOR signaling pathway,inhibition of endothelial au-tophagy and apoptosis,and promotion of angiogenesis in diabetic rats.关键词
糖尿病/皮肤创伤/硫化氢/自噬/血管形成/PI3K/Akt/mTOR信号通路Key words
diabetes mellitus/skin wound/hydrogen sulfide/autophagy/angiogenesis/PI3K/Akt/mTOR signaling pathway分类
临床医学引用本文复制引用
李媛媛,赵富生,张可心,陈永兰,张娜,姜昕悦,谷春付,武庚..硫化氢对糖尿病大鼠皮肤创面自噬和血管形成的影响[J].中国病理生理杂志,2023,39(12):2223-2233,11.基金项目
黑龙江省自然科学基金项目(No.LH2020H073) （No.LH2020H073）
黑龙江省省属高等学校基本科研业务费项目(No.2019-KYY-WFMY-0019) （No.2019-KYY-WFMY-0019）
红旗科研基金科技项目(No.2018-HQ-04) （No.2018-HQ-04）
牡丹江医学院科学基金火炬计划(No.2022-MYHJ-005) （No.2022-MYHJ-005）
