CAJ | 학술논문

目的：探讨外源性硫化氢( H2 S)对压疮缺血/再灌注( I/R)损伤大鼠的保护作用及其相关机制。方法将40只SD大鼠按随机数字表法分为4组：正常对照组、I/R模型组及低、高剂量H2 S干预组,每组10只。低、高剂量H2 S干预组分别给予10、30μmol/kg的硫氢化钠( NaHS)腹腔注射1周,正常对照组与模型组接受等量的9 g/L盐水,然后正常对照组不作受压处理,其余3组均于压疮装置上受压,3个I/R循环后处死大鼠,通过 HE 染色判断压疮病理改变程度,利用亚甲基分光光度法分析血清 H2 S 水平,行酶联免疫吸附法( ELISA)检测血清肿瘤坏死因子-α( TNF-α)、白细胞介素-6( IL-6)、细胞间黏附分子-1( ICAM-1)、髓过氧化物酶( MPO)水平,测定肌肉匀浆液中丙二醛( MDA)、超氧化物歧化酶( SOD)、谷胱甘肽过氧化物酶( GSH-Px)、过氧化氢酶( CAT)活性,采用原位切口末端标记法( TUNEL)计算肌细胞凋亡指数( AI),Western blot分析肌肉组织Bcl-2、Bax表达情况。结果与正常对照组比较,I/R模型组血清TNF-α、IL-6、ICAM-1、MPO水平、肌肉匀浆液中MDA活性、AI及肌肉组织 Bax 表达水平增加(P均<0.01),血清H2S水平、肌肉匀浆液SOD、GSH-Px、CAT活性及肌肉组织 Bcl-2表达水平降低(P均<0.01)。与I/R模型组比较,经低、高剂量H2S干预治疗后,病理积分、血清TNF-α、IL-6、ICAM-1、MPO水平、肌肉匀浆液中MDA活性、AI及肌肉组织 Bax 表达水平逐渐减少( P均<0．05、0.01),血清H2S水平、肌肉匀浆液SOD、GSH-Px、CAT活性及肌肉组织 Bcl-2表达水平逐渐增高(P均<0．05、0.01)。结论外源性H2 S对压疮I/R损伤大鼠产生良好的保护作用,其机制可能与抑制炎性反应、增强抗氧化能力及减少细胞凋亡有关。
목적：탐토외원성류화경( H2 S)대압창결혈/재관주( I/R)손상대서적보호작용급기상관궤제。방법장40지SD대서안수궤수자표법분위4조：정상대조조、I/R모형조급저、고제량H2 S간예조,매조10지。저、고제량H2 S간예조분별급여10、30μmol/kg적류경화납( NaHS)복강주사1주,정상대조조여모형조접수등량적9 g/L염수,연후정상대조조불작수압처리,기여3조균우압창장치상수압,3개I/R순배후처사대서,통과 HE 염색판단압창병리개변정도,이용아갑기분광광도법분석혈청 H2 S 수평,행매련면역흡부법( ELISA)검측혈청종류배사인자-α( TNF-α)、백세포개소-6( IL-6)、세포간점부분자-1( ICAM-1)、수과양화물매( MPO)수평,측정기육균장액중병이철( MDA)、초양화물기화매( SOD)、곡광감태과양화물매( GSH-Px)、과양화경매( CAT)활성,채용원위절구말단표기법( TUNEL)계산기세포조망지수( AI),Western blot분석기육조직Bcl-2、Bax표체정황。결과여정상대조조비교,I/R모형조혈청TNF-α、IL-6、ICAM-1、MPO수평、기육균장액중MDA활성、AI급기육조직 Bax 표체수평증가(P균<0.01),혈청H2S수평、기육균장액SOD、GSH-Px、CAT활성급기육조직 Bcl-2표체수평강저(P균<0.01)。여I/R모형조비교,경저、고제량H2S간예치료후,병리적분、혈청TNF-α、IL-6、ICAM-1、MPO수평、기육균장액중MDA활성、AI급기육조직 Bax 표체수평축점감소( P균<0．05、0.01),혈청H2S수평、기육균장액SOD、GSH-Px、CAT활성급기육조직 Bcl-2표체수평축점증고(P균<0．05、0.01)。결론외원성H2 S대압창I/R손상대서산생량호적보호작용,기궤제가능여억제염성반응、증강항양화능력급감소세포조망유관。
Objective To explore the protective effect and mechanism of exogenous hydrogen sulfide ( H2 S) on pressure ulcer rats with ischemia/reperfusion ( I/R) injury. Methods Forty SD rarts were randomly divided into 4 groups,namely normal control group,I/R model group,low-dose H2 S intervention group and high-dose H2 S interven-tion group. Each group had 10 animals. The low-and the high-dose H2 S intervention groups were injected intraperitone-ally 10 and 30μmol/kg of sodium hydrosulfide ( NaHS) for 1 week,respectively;however,the normal control group and I/R model group were treated with same volume 9 g/L saline. Then,the normal control group did not sustain any pres-sure,but other 3 groups were pressurized. After 3 circulations of ischemia/reperfusion,all rats were sacrificed,and path-ological changes in pressure ulcer rats were evaluated by HE staining. Serum H2 S contents were analyzed through meth-ylene spectrophotometric method. Serum levels of tumor necrosis factor-α(TNF-α),interleukin-6 (IL-6),intercellu-lar adhesion molecule -1 (ICAM-1),and myeloperoxidase (MPO) were detected by enzyme linked immunosorbent assay ( ELISA ) . The activities of malondialdehyde ( MDA ) , superoxide dismutase ( SOD ) , glutathione-peroxidase ( GSH-Px) and catalase ( CAT) in the homogenate of muscle were examined. Apoptotic index ( AI) was calculated by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling ( TUNEL) . The expressions of Bcl-2 and Bax in muscle tissue were evaluated by Western blot. Results In comparison with the normal control group,serum TNF-α, IL-6,ICAM-1 and MPO levels,MDA activity in the homogenate of muscles,AI and Bax expression level were in-creased (all P<0. 01),while serum H2S levels,the activities of SOD,GSH-Px and CAT as well as Bcl-2 expression levels were reduced in the I/R model group (all P<0. 01). Upon intervention therapy of low-or high-dose H2S,path-ological scores,serum TNF-α,IL-6,ICAM-1 and MPO levels,MDA activity in the homogenate of muscle,AI and Bax expression levels were gradually reduced while serum H2 S levels,the activities of SOD,GSH-Px and CAT and Bcl-2 expression levels were gradually enhanced when compared to those of the I/R model group (all P<0. 05,0. 01). Con-clusions Exogenous H2 S has better protection against pressure ulcer rats with I/R injury. The mechanism may be as-sociated with inhibition of inflammatory reaction and enhancement of antioxidative ability as well as reduction of apopto-sis.