Zhao Zi-gang, Niu Chun-yu, Zhang Yu-ping, Du Shu-ting, Zhang Jing
Department of Pathophysiology, Hebei Northern University, Zhangjiakou 075000, Hebei, China.
Zhongguo Wei Zhong Bing Ji Jiu Yi Xue. 2010 Feb;22(2):97-100.
To explore the effect of mesenteric lymph duct ligation in protecting myocardial injury in rats after hemorrhage and lipopolysaccharide (LPS) challenge.
Wistar rats were randomly divided into the sham group, non-ligation group and ligation group, and the "two-hit" injury model was reproduced by hemorrhage and LPS administration. Mesenteric lymph was blocked by ligating mesenteric lymph duct in ligation group after hemorrhage. After 24 hours of the assault, myocardial tissue was harvested and homogenized. The activity of myeloperoxidase (MPO) and ATPase and the contents of tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) were determined in myocardial homogenate. At the same time, myocardium was pathologically studied. Apoptosis cell rate of myocardium was determined by method of terminal-deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL), the expression of bcl-2 and bax protein was determined by immunohistochemical method.
After hemorrhage and LPS challenge, the level of MPO [(0.23+/-0.08) U/g], TNF-alpha [(9.99+/-2.74) microg/g] and IL-6 [(31.57+/-12.71) microg/g] in myocardial homogenate of non-ligation group were significantly higher than those of sham group [MPO: (0.12+/-0.03) U/g, TNF-alpha: (4.17+/-1.35) microg/g, IL-6: (17.86+/-5.17) microg/g, respectively, all P<0.01], and the activity of ATPase was significantly lower. MPO [(0.13+/-0.03) U/g], TNF-alpha [(5.57+/-1.65) microg/g] and IL-6 [(23.24+/-5.95) microg/g] in myocardial homogenate of ligation group were significantly lower (P<0.05 or P<0.01), and the ATPase activity was significantly higher compared with non-ligation group. The apoptosis rate [(22.7+/-6.9)%] and expression of bax protein (104.5+/-11.4) of myocardial cells in non-ligation group were significantly higher compared with sham group [apoptosis rate: (3.8+/-1.2)%, bax protein: 142.1+/-10.9] and ligation group [apoptosis rate: (8.4+/-2.8)%, bax protein: 128.4+/-9.6], and expression of bcl-2 protein of non-ligation group (196.4+/-19.3) was significantly lower than that of sham group (132.2+/-12.3) and ligation group (165.1+/-11.6, all P<0.01).
The results demonstrate that the mesenteric lymph duct ligation ameliorate the myocardial injury in rats subjected to hemorrhage and LPS by reducing the inflammatory mediators of TNF-alpha and IL-6 levels, upregulating the bcl-2 protein expression and improving membrane ATPase activity of myocardium.
探讨肠系膜淋巴管结扎对出血及脂多糖(LPS)攻击后大鼠心肌损伤的保护作用。
将Wistar大鼠随机分为假手术组、未结扎组和结扎组,通过出血及给予LPS复制“两次打击”损伤模型。出血后在结扎组结扎肠系膜淋巴管以阻断肠系膜淋巴。攻击24小时后,取心肌组织并匀浆。测定心肌匀浆中髓过氧化物酶(MPO)、ATP酶活性以及肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)的含量。同时,对心肌进行病理研究。采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法(TUNEL)测定心肌凋亡细胞率,用免疫组织化学方法测定bcl-2和bax蛋白的表达。
出血及LPS攻击后,未结扎组心肌匀浆中MPO[(0.23±0.08)U/g]、TNF-α[(9.99±2.74)μg/g]和IL-6[(31.57±12.71)μg/g]水平显著高于假手术组[MPO:(0.12±0.03)U/g,TNF-α:(4.17±1.35)μg/g,IL-6:(17.86±5.17)μg/g,均P<0.01],且ATP酶活性显著降低。结扎组心肌匀浆中MPO[(0.13±0.03)U/g]、TNF-α[(5.57±1.65)μg/g]和IL-6[(23.24±5.95)μg/g]显著降低(P<0.05或P<0.01),与未结扎组相比,ATP酶活性显著升高。未结扎组心肌细胞凋亡率[(22.7±6.9)%]和bax蛋白表达(104.5±11.4)显著高于假手术组[凋亡率:(3.8±1.2)%,bax蛋白:142.1±10.9]和结扎组[凋亡率:(8.4±2.8)%,bax蛋白:128.4±9.6],未结扎组bcl-2蛋白表达(196.4±19.3)显著低于假手术组(132.2±12.3)和结扎组(165.1±11.6,均P<0.01)。
结果表明,肠系膜淋巴管结扎通过降低TNF-α和IL-6水平等炎症介质、上调bcl-2蛋白表达及提高心肌膜ATP酶活性,改善出血及LPS攻击大鼠的心肌损伤。