作为气体分子的硫化氢(H2S)防止胃肠道(GI)对各种损伤。本研究旨在首次评估线粒体靶向H2S前药的详细分子机制,AP39和RT01对缺血/再灌注(I/R)诱导的损伤的胃保护作用。对暴露于I/R的Wistar大鼠进行i.g.AP39(0.004-2mg/kg),RT01(0.1mg/kg),或使用AP219(0.1mg/kg)作为结构控制,而没有释放H2S的能力。AP39还与mTOR1抑制剂一起施用,雷帕霉素(1mg/kgi.g.)。胃损伤面积进行了微观/宏观评估,通过激光流量计测量胃血流量(GBF),HIF-1αmRNA水平,GPx,SOD1,SOD2,膜联蛋白A1,SOCS3,IL-1RA,IL-1β,IL-1R1、IL-1R2、TNFR2、iNOS经实时PCR检测。胃粘膜和/或血清IL-1β含量,IL-4,IL-5,IL-10,G-CSF,M-CSF,VEGFA,GRO,RANTES,MIP-1α,MCP1,TNF-α,TIMP1,FABP3,GST-α,STAT3/5和mTOR的磷酸化,NF-κB,ERK,通过微珠-荧光测定评价Akt。用生物化学方法测量线粒体复合物的活性。通过ELISA将RNA损伤评估为8-OHG。AP39和RT01减少了增加GBF的微观/宏观胃I/R损伤。AP39-胃保护伴随着线粒体复合物的维持活性,防止RNA氧化并增强SOCS3,IL-1RA的mRNA/蛋白表达,膜联蛋白A1,GST-α,HIF-1α。雷帕霉素逆转了AP-39-胃保护。AP39-胃保护作用后NF-κB降低,ERK,IL-1β和增强Akt和mTOR蛋白磷酸化。AP39预防由I/R损伤引起的胃粘膜损伤,部分由线粒体复合物活性维持。AP39介导的胃粘膜氧化衰减,缺氧和炎症涉及mTOR1和Akt通路的活性和HIF-1α的调节,GST-α,SOCS3、IL1RA和TIMP1分子相互作用。
Hydrogen sulfide (H2S) as a gaseous molecule prevents gastrointestinal (GI)-tract against various injuries. This study aimed to evaluate for the first time the detailed molecular mechanism of mitochondria-targeting H2S-prodrugs, AP39 and RT01 in gastroprotection against ischemia/reperfusion (I/R)-induced lesions. Wistar rats exposed to I/R were pretreated i.g. with vehicle, AP39 (0.004-2 mg/kg), RT01 (0.1 mg/kg), or with AP219 (0.1 mg/kg) as structural control without ability to release H2S. AP39 was also administered with mTOR1 inhibitor, rapamycin (1 mg/kg i.g.). Gastric damage area was assessed micro-/macroscopically, gastric blood flow (GBF) by laser flowmetry, mRNA level of HIF-1α, GPx, SOD1, SOD2, annexin-A1, SOCS3, IL-1RA, IL-1β, IL-1R1, IL-1R2, TNFR2, iNOS by real-time PCR. Gastric mucosal and/or serum content of IL-1β, IL-4, IL-5, IL-10, G-CSF, M-CSF, VEGFA, GRO, RANTES, MIP-1α, MCP1, TNF-α, TIMP1, FABP3, GST-α, STAT3/5 and phosphorylation of mTOR, NF-κB, ERK, Akt was evaluated by microbeads-fluorescent assay. Mitochondrial complexes activities were measured biochemically. RNA damage was assessed as 8-OHG by ELISA. AP39 and RT01 reduced micro-/macroscopic gastric I/R-injury increasing GBF. AP39-gastroprotection was accompanied by maintained activity of mitochondrial complexes, prevented RNA oxidation and enhanced mRNA/protein expression of SOCS3, IL-1RA, annexin-A1, GST-α, HIF-1α. Rapamycin reversed AP-39-gastroprotection. AP39-gastroprotection was followed by decreased NF-κB, ERK, IL-1β and enhanced Akt and mTOR proteins phosphorylation. AP39-prevented gastric mucosal damage caused by I/R-injury, partly by mitochondrial complex activity maintenance. AP39-mediated attenuation of gastric mucosal oxidation, hypoxia and inflammation involved mTOR1 and Akt pathways activity and modulation of HIF-1α, GST-α, SOCS3, IL1RA and TIMP1 molecular interplay.