富氢液对脓毒血症大鼠肠黏膜屏障保护作用及机制研究

目的探讨富氢液对脓毒血症大鼠肠黏膜屏障的影响及其可能机制。方法清洁级SD大鼠30只,随机分为假手术组(sham组)、脓毒血症组(SEP组)及富氢液处理组(C组),每组10只。sham组仅开腹,显露盲肠;C组和SEP组行盲肠结扎穿孔术,再分别静脉给予富氢液和等量生理盐水。3组大鼠均于模型建立4 h后处死,观察小肠组织病理学结果;酶联免疫吸附法(ELISA)检测肠脂肪酸结合蛋白(I-FABP)、D-乳酸、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-10(IL-10)浓度;蛋白质印迹(Western-blot)法检测B淋巴细胞瘤-2基因(Bcl-2)、Bax、Caspase-3蛋白表达水平。结果光镜下sham组未出现病理损伤,SEP组损伤较重,C组损伤较轻。与sham组比较,SEP组I-FABP、D-乳酸升高(817.12±50.39)ng/ml比(378.54±40.50)ng/ml;(513.84±42.91)ng/ml比(329.98±38.11)ng/ml,P<0.05]。与SEP组比较,C组I-FABP、D-乳酸降低(548.92±42.19)ng/ml比(817.12±50.39)ng/ml;(437.89±36.12)ng/ml比(513.84±42.91)ng/ml,P<0.05]。与sham组比较,SEP组TNF-α、IL-6、IL-10升高(196.51±30.31)ng/ml比(75.54±10.21)ng/ml;(68.69±3.39)ng/ml比(20.97±2.38)ng/ml;(149.61±20.21)ng/ml比(80.69±13.51)ng/ml,P<0.05];与SEP组比较,C组TNF-α、IL-6降低,IL-10升高(118.47±10.14)ng/ml比(196.51±30.31)ng/ml;(45.36±5.14)ng/ml比(68.69±3.39)ng/ml;(178.52±26.31)ng/ml比(149.61±20.21)ng/ml,P<0.05]。与sham组比较,SEP组Bcl-2蛋白表达升高,Bax与Caspase-3降低(P<0.05);与SEP组比较,C组Bcl-2蛋白表达降低,Bax与Caspase-3升高(P<0.05)。结论富氢液对脓毒血症大鼠肠黏膜屏障有保护作用,其机制与抑制凋亡相关因子有关。

Protective effect and mechanism of hydrogen-rich saline in sepsis of intestinal mucosa barrier in rats

Objective To investigate the effect of hydrogen-rich saline in septic rat intestinal mucosa injury and the expression of ap-optosis correlated factors. Methods A total of 30 SD rats in clean grade were randomly divided into the sham operation group( Group Sham),the sepsis group(Group SEP) and the hydrogen-rich saline treatment group(Group C),10 rats in each group. Rats of the Group Sham were only exposed to receive laparotomy,the other rats of Group C and Group SEP received cecum ligation and puncture, and were given intravenous hydrogen-rich saline and the same amount of saline,respectively. Three groups of model rats were sacrificed after 4 hours,all the detections were observed such as the intestinal pathological detection;ELISA detection of blood intestinal fatty acid binding protein(I-FABP),D-concentration of lactic acid,tumor necrosis factor-α(TNF-α),concentration of interleukin-6(IL-6) and interleukin-10 ( IL-10 );Western-blot detection of B-cell lymphoma ( Bcl-2 ) , Bax and Caspase-3 protein expression level. Results Under light microscope,there was no pathological damage in the Group Sham,severe damage in the Group SEP and lighter damage in the Group C. Compared with the Group Sham,I-FABP and D-concentration of lactic acid were increased in the Group SEP (817. 12 ±50. 39) ng/ml versus (378. 54 ± 40. 50) ng/ml;(513. 84 ± 42. 91) ng/ml versus (329. 98 ± 38. 11) ng/ml,P <0. 05]. Compared with the Group SEP,I-FABP and D-concentration of lactic acid were decreased in the Group C(548. 92 ±42. 19) ng/ml versus (817. 12 ± 50. 39) ng/ml;(437. 89 ± 36. 12) ng/ml versus (513. 84 ± 42. 91) ng/ml,P<0. 05]. Compared with the Group Sham,TNF-α,IL-6 and IL-10 were increased in the Group SEP(196. 51 ± 30. 31) ng/ml versus (75. 54 ± 10. 21) ng/ml;(68. 69 ± 3. 39) ng/ml versus (20. 97 ± 2. 38) ng/ml;(149. 61 ± 20. 21) ng/ml versus (80. 69 ± 13. 51) ng/ml,P<0. 05];compared with the Group SEP, TNF-α and IL-6 were decreased, while IL-10 was increased in the Group C ( 118. 47 ± 10. 14 ) ng/ml versus (196. 51 ±30. 31) ng/ml;(45. 36 ± 5. 14) ng/ml versus (68. 69 ± 3. 39) ng/ml;(178. 52 ± 26. 31) ng/ml versus (149. 61 ± 20. 21) ng/ml,P<0. 05]. Compared with the Group Sham,protein expression of Bcl-2 was increased,while those of Bax and Caspase-3 were decreased in the Group SEP(P<0. 05);compared with the Group SEP,protein expression of Bcl-2 was decreased,while those of Bax and Caspase-3 were increased in the Group C(P<0. 05). Conclusion Hydrogen-rich saline has protective effect on intestinal mucosal barrier in rats with sepsis,and its mechanism is related to inhibition of apoptosis relative factors.