CAJ | 학술논문

目的探讨盐酸戊乙奎醚后处理对肢体缺血再灌注后肾脏的保护作用及其机制.方法健康成年雄性Wistar大鼠72只,体重220～250 g,随机数字表法分为3组:对照组(C组)、肢体缺血再灌注组(I/R组)、盐酸戊乙奎醚后处理组(P组),根据缺血后再灌注时间点各组又分为缺血3 h(T0)、再灌注1 h(T1)、3 h(T2)、6 h(T3)四个亚组(n=6).除C组外,各组在大鼠双后肢根部用橡皮筋结扎,完全阻断血流3 h.P组在再灌注前3 min尾静脉注射盐酸戊乙奎醚0.15 mg/kg(0.8ml).比色法检测血清肌酐(Cr)和尿素氮(BUN)的含量、肾脏组织超氧化物歧化酶(SOD)的活性及丙二醛(MDA)的含量,酶联免疫吸附法测定血清肿瘤坏死因子α(TNF-α)的含量,免疫组织化学SABC法测定肾脏组织中缺氧诱导因子1α(HIF-1α)的表达,光镜下观察肾脏组织的病理学改变.结果 I/R组和P组血清BUN、Cr水平、SOD的活性、MDA水平、TNF-α水平、HIF-1α的表达均高于C组(均P<0.05);P组血清BUN、Cr、MDA、TNF-α水平及、HIF-1α表达均低于I/R组[T2时间点:(15.10 ±1.88)mmol/L比(19.46±2.76)mmol/L、(113±10)μmol/L比(143±11)μmol/L、(13.8 ±1.7)nmol/g比(15.5±1.8)nmol/g、(53.1±3.1)ng/L比(53.9±4.8)ng/L、0.298±0.015比0.471±0.032,均P<0.05],SOD的活性高于I/R组(P<0.05).结论盐酸戊乙奎醚后处理可以下调HIF-1α的表达,减轻肢体缺血再灌注后肾脏的损伤.其机制可能是抑制了炎症反应及氧自由基的释放,改善了肾脏组织的缺血、缺氧状态.
목적 탐토염산무을규미후처리대지체결혈재관주후신장적보호작용급기궤제.방법 건강성년웅성Wistar대서72지,체중220～250 g,수궤수자표법분위3조:대조조(C조)、지체결혈재관주조(I/R조)、염산무을규미후처리조(P조),근거결혈후재관주시간점각조우분위결혈3 h(T0)、재관주1 h(T1)、3 h(T2)、6 h(T3)사개아조(n=6).제C조외,각조재대서쌍후지근부용상피근결찰,완전조단혈류3 h.P조재재관주전3 min미정맥주사염산무을규미0.15 mg/kg(0.8ml).비색법검측혈청기항(Cr)화뇨소담(BUN)적함량、신장조직초양화물기화매(SOD)적활성급병이철(MDA)적함량,매련면역흡부법측정혈청종류배사인자α(TNF-α)적함량,면역조직화학SABC법측정신장조직중결양유도인자1α(HIF-1α)적표체,광경하관찰신장조직적병이학개변.결과 I/R조화P조혈청BUN、Cr수평、SOD적활성、MDA수평、TNF-α수평、HIF-1α적표체균고우C조(균P<0.05);P조혈청BUN、Cr、MDA、TNF-α수평급、HIF-1α표체균저우I/R조[T2시간점:(15.10 ±1.88)mmol/L비(19.46±2.76)mmol/L、(113±10)μmol/L비(143±11)μmol/L、(13.8 ±1.7)nmol/g비(15.5±1.8)nmol/g、(53.1±3.1)ng/L비(53.9±4.8)ng/L、0.298±0.015비0.471±0.032,균P<0.05],SOD적활성고우I/R조(P<0.05).결론 염산무을규미후처리가이하조HIF-1α적표체,감경지체결혈재관주후신장적손상.기궤제가능시억제료염증반응급양자유기적석방,개선료신장조직적결혈、결양상태.
Objective To evaluate the protection of penehyclidine hydrochloric postconditioning on HIF-1α (hypoxia-inducible factor -1α) in renal tissue injury induced by lower limb ischemia/reperfusion (I/R). Methods A total of 72 adult male Wistar rats weighing 230 - 250 g were randomly divided into 3 groups: control ( group C ) , limb ischemia-reperfusion ( group R/I) and penehyclidine hydrochloride postconditioning (group P). The animals were anesthetized by inhaling 2% isoflurane and blood flow of bilateral lower limbs was blocked with rubber bands for 3 h in groups P and R/I. In group P, penehyclidine hydrochloride 0. 15 mg/kg was injected via caudal vein at 3 min pre-reperfusion. After sacrificing, their kidneys were removed at 3 h of ischemia and 1, 3, 6 h of reperfusion respectively. The blood urea nitrogen (BUN) and creatinine ( Cr) were detected by colorimetric method, plasma tumor necrosis factor-α (TNF-α) by ELISA ( enzyme-linked immunosorbent assay ) and HIF-1α of renal tissue by immunohistochemistry. Renal pathological changes were observed under light microscope. Results Compared with group C, the serum levels of BUN and Cr increased while TNF-α and HIF-1α were upregulated in groups I/R and P (P < 0. 05). As compared with group I/R, the serum levels of BUN, Cr and MDA decreased while TNF-α and HIF-lα were down-regulated in group P . [at T2: (15. 10 ± 1. 88) mmol/L vs(19.46±2. 76) mmol/L, (113 ±10) μmol/L vs(143 ± 11) μmol/L, (13. 8 ±1.7) nmol/g vs (15.5 ±1.8) nmol/g, (53.1 ±3. 1)ng/L vs(53.9 ±4. 8) ng/L, 0.298 ±0.015 vs 0.471 ±0.032, all P<0.05 ]. Conclusion Penehyclidine hydrochloride can down-regulate the expression of HIF-lα and attenuate the renal injury induced by lower limb I/R. And the mechanisms may be through inhibiting the inflammatory reactions, reducing the release of oxygen free radicals and improving the conditions of hypoxia and ischemia.