核因子-κB抑制剂对孕期感染鼠子代神经发育缺陷的干预研究

目的 探讨孕期感染鼠母体细胞因子介导的免疫激活与子代神经发育缺陷之间的关系以及核因子-κB抑制剂的干预效果.方法 60只大鼠随机分为三组,40只大鼠在孕早期应用Poly(I:C)制造孕鼠感染模型,每20只分别为模型组和干预组,干预组应用NF-κB抑制剂吡咯烷二硫基甲酸盐(PDTC)阻断相关的细胞因子表达,20只大鼠给予安慰剂注射作为对照.每组半数处死ELISA法检测母体血清IL-10和TNF-α水平,其余完成妊娠,对不同处理组孕鼠后代在成年期比较前脉冲抑制、被动回避和主动规避的不同.结果 应用Poly(I:C)后,母体血清IL-10[对照组(0.16±0.13)pg/ml比模型组(18.26±1.52)pg/m1]和TNF-α对照组[(11.21±1.81)pg/ml比模型组(119.64±16.42)pg/m1]水平升高,同时子代成年期出现精神病样表现如前脉冲抑制、被动回避和主动规避的异常,PDTC可以降低母体血清IL-10[干预组(12.64±2.04)pg/ml比模型组(18.26±1.52)pg/m1]和TNF-α[干预组(30.34±2.19)pg/ml比模型组(119.64±16.42)pg/ml]水平,部分改善子代成年期出现的精神病样行为.结论 PDTC通过降低孕鼠感染后细胞因子介导的母体炎性反应,可干预子代成年后的神经发育障碍.

Abstract：

Objective To explore the correlation between the elevated expression of cytokines under offsprings and understand the intervening effects of nuclear factor NF-κB inhibitor pyrrolidinedithiocarbamate pregnancy in rats. And the expression of cytokines was blocked with PDTC. The maternal levels of tumor necrosis factor alpha and intcrleukin-10 were determined by ELISA (enzyme-linked immunosorbent assay).The adult offsprings on different treatments were then compared with regards to prepulse inhibition ( PPI ),elevated levels of serum cytokines as shown by the markedly increased serum levels of IL-10 and TNF-α.The serum levels of IL-10 and TNF-α in the model group were significantly higher than those in the control group [(18.26±1.52) pg/ml, (119.64±16.42) pg/ml vs. (0. 16±0. 13) pg/ml and (11.21 ±1.81)pg/ml]. The elevation was partly blocked by PDTC. The serum levels of IL-10 and TNF-α in the intervention group [ ( 12. 64 ±2.04) pg/ml and (30. 34 ±2. 19) pg/ml respectively] were lower than those in the model group, but still higher than those in the control group. The psychotic-like phenotypes including defects in was blunted by the PDTC intervention. The PPI results demonstrated that the PP2 and PP8 difference between rats in 3 groups were statistically significant, with a lower PPI value in the model group than in the intervention group, in the intervention group than in the control group and much lower in the model group than in the control group. PP4 was lower in the model group than that in the intervention group, and also lower in the model group than in the control group. There was no significant difference between the control group and the intervention group. The passive avoidance results indicated that T1 was higher in the model group than in the control and intervention groups and there was no statistical difference between the control and intervention groups. T2 was lower in the model group than in the control and intervention groups and there was no statistical difference between the control and intervention groups. And the active avoidance test results showed that total conditioned reflex times of the control group was higher than those of the intervention and model groups. No statistical difference was found between the intervention and model groups. Total reflex rate of the control group was higher than that of the intervention and model groups. No statistical difference was found between the intervention and model groups. Conclusion PDTC can interfere with neural developmental disorder of adult offsprings through blunting the cytokine-mediated maternal immune response.     

Protective effects of hemin pretreatment combined with ulinastatin on septic shock in rats

Background Urinary trypsin inhibitor inhibits the enhanced production of pro-inflammatory molecules. Hemeoxygenase-1 induction protects against ischemia/repeffusion injury, oxidative stress, inflammation, transplant rejection, apoptosis, and other conditions. However, it is unknown if a combined hemin and ulinastatin pretreatment could result in protective effects for septic shock. In this study, we investigated the role of hemin pretreatment combined with ulinastatin on septic shock in rats. Methods Eighty healthy, male Sprague-Dawley rats were randomly divided into four groups： group S, group H, group U and group HU. Groups S and U received 1 ml normal saline intraperitoneally, while groups H and HU both received 1 ml （100 mg/kg） hemin. Twenty-four hours later, 0.5 ml （10 mg/kg） E. coil lipopolysaccharide was injected intravenously to replicate the experimental model of septic shock. After an initial 25% decrease in the mean arterial pressure, corresponding to time point 0, groups HU and U received 0.5 ml 10 000 U/kg ulinastatin intravenously, and the others received 0.5 ml normal saline. Results The number of deaths in groups H and U was lower than that in the group S （P〈0.05）, and was higher than that in group HU （all P〈0.05） respectively. The mean arterial pressure （MAP） in the group S was significantly greater than that in group H （P〈0.05）, and was lower than that in group HU and group U （P〈0.05）. The plasma levels of alanine aminotransferase （ALT）, aspartate aminotransferase （AST）, creatinine （Cr） and blood urea nitrogen （BUN）, the malondial- dehyde （MDA） of liver, kidney and lung, and the lung Evans blue （EB） contents in groups H and U, were greater than that in group HU （all P〈0.05）, and were lower than that in group S （all P〈0.05）. In contrast, the plasma levels of CO in groups H and HU were higher than that in groups S and U （all P〈0.05）, and SOD of liver, kidney and lung in groups H and U were higher than that in group S,     

淋巴引流及ω-3多不饱和脂肪酸干预对大鼠肠道缺血再灌注的影响

目的 观察大鼠肠道缺血再灌注(Ⅰ/R)损伤时肠淋巴液引流对高迁移率族蛋白1(HMGB1)、炎症因子和内毒素的影响以及ω-3多不饱和脂肪酸(ω-3 PUFA)干预的效果.方法 72只SD大鼠随机区组法随机分为单纯引流组、Ⅰ/R组、Ⅰ/R+引流组(每组8只)和胃造口组[正常饮食(N)组、普通肠内营养(EN)组、普通肠内营养加ω-3 PUFA(PUFA)3大组,每大组再根据是否行Ⅰ/R 和引流分为2组,每组8只].单纯引流组只引流180 mⅠn淋巴液不行Ⅰ/R损伤;Ⅰ/R、VR+引流组行肠系膜上动脉夹闭60 mⅠn再灌注120 mⅠn;Ⅰ/R+引流组同时行肠淋巴液引流180 mⅠn.胃造口组大鼠均先行胃造口手术,分别给予不同营养5 d后造模,各引流组同前进行肠淋巴液引流180 mⅠn.手术完毕后分别取血清和淋巴液,定量检测内毒素,酶联免疫吸附试验(ELⅠSA)定量检测炎症因子以及HMGB1.结果 Ⅰ/R+引流组淋巴液中内毒素、炎症因子以及HMGBl均高于单纯引流组[均P＜0.05,白细胞介素(ⅠL)-6(30±8)pg/ml比(20±6)pg/ml,内毒素(0 029±0.011)U/ml比(0 008±0 005)U/ml];Ⅰ/R+引流组血清中内毒素、炎症因子均低于Ⅰ/R组(均P＜0 05).在胃造口组中,N 组和EN组的淋巴液中肿瘤坏死因子(TNF)-α与HMGBl均高于PUFA组[(46±17)pg/ml、(54±16)pg/ml比(28±9)pg/ml,(4.8±1.6)ng/ml、(5.3±1.8)ns/ml比(3.0±1.0)ng/ml,均P＜0.05].PUFA(Ⅰ/R)组血清中内毒素、炎症因子以及HMGBl均低于N(Ⅰ/R)组(均P＜0.05),PUFA(Ⅰ/R+引流)组血清中TNF-α与HMGBl均低于N(Ⅰ/R+引流)组(均P＜0 05).结论 引流肠淋巴液能够降低肠道Ⅰ/R损伤时内毒素、炎症因子和HMGB1的水平,减轻大鼠肠道Ⅰ/R引起的损伤.ω-3PUFA的干预对于肠道Ⅰ/R引起的损伤有一定的保护作用,对于减轻炎症反应有积极作用.

Abstract：

Objective To investigate the effects of lymphatic drainage and ω-3 polyunsaturated fatty acid (ω-3PUFA) on high mobility group box 1 (HMGB1) , inflammatory cytokines and endotoxin in rats with intestinal ischemia-reperfusion (Ⅰ/R) injury. Methods A total of 72 SD rats were randomly divided into drainage-alone group, Ⅰ/R group, ischemia-reperfusion plus drainage (Ⅰ/R+D) group (n=8 each)and 3 groups with 16 rats undergoing gastrostomy in each group: normal diet (N) group, enteral nutrition (EN) group and enteral nutrition & ω-3PUFA (PUFA) group. And they were further divided into 2 subgroups (n=8). The rats in Ⅰ/R and Ⅰ/R+D groups were subjected to a 60-min ischemia follow by 120-min reperfusion injury of superior mesenteric artery. When the rats suffered Ⅰ/R injury, intestinal lymph was drained for 180 min in the Ⅰ/R+D group. The rats in the drainage-alone group received 180-min lymph drainage without Ⅰ/R injury. After 5 days with different nutrition regimes, the models were established similarly. The rats in the Ⅰ/R+D sub-groups were treated with intestinal lymph drainage for 180 min. The serum and lymph samples were collected post-operatively. Endotoxin was detected by a Limulus kit. The inflammatory cytokines and high mobility group box 1 (HMGB1) were analyzed by enzyme-linked immunosorbent assay (ELISA).Results Endotoxin, inflammatory cytokines and lymphatic HMGB1 of lymphatic in the Ⅰ/R+D group were higher than those in the drainage-alone group [all P＜0.05 , IL-6 :(30±8) pg/ml vs (20±6) pg/ml, endotoxin: (0.029±0.011) U/ml vs (0.008+0.005) U/ml].The serum levels of endotoxin and inflammatory cytokines in the Ⅰ/R+ D group were lower than those in the Ⅰ/R group (P＜0.05).The lymphatic levels of TNF-a (tumor necrosis factor-alpha) and HMGB1 in the N and EN groups were higher than those in the PUFA group[TNF-α: (46±17)pg/ml, (54±16)pg/ml vs(28±9) pg/ml, HMGB1: (4.8±1.6) ng/ml, (5.3±1.8) ng/ml, (3.0±1.0) ng/ml, all P＜0.05) ].The serum levels of endotoxin, inflammatory cytokines and HMGB1 in the PUFA(l/R) group were lower than those in the N(Ⅰ/R) group (F＜0.05).The levels of TNF-a and HMGB1 were lower in the PUFA (Ⅰ/R+D) group than those in the N(Ⅰ/R+ D) group (both P＜0.05).Conclusion Lymphatic drainage may reduce the levels of endotoxin, inflammatory cytokines and HMGB1 so as to alleviate the intestinal Ⅰ/R injury. The intervention of ω-3PUFA has some protective effect through relieving inflammation.     

异丙酚对新生大鼠空间学习记忆功能及内源性神经干细胞增殖的影响

目的 探讨不同剂量的异丙酚对新生大鼠空间学习记忆功能发育及对内源性神经干细胞增殖的影响.方法 5窝新生SD大鼠按照随机区组化分为对照组(C组)、3种不同剂量异丙酚组(P10组、P50组、P50D组),每组15只.P10、P50组分别单次皮下注射异丙酚10mg/kg、50mg/kg;P50D组先后2次皮下注射异丙酚(50mg/kg);C组注射等容量的脂肪乳剂.3d后用BrdU法检测海马齿状回神经元的增殖;使用水迷宫检测出生后28d的幼鼠空间学习记忆能力.结果 免疫组化的检测中,P10组齿状回单位面积含BrdU阳性细胞的数量[(1225±154)个/mm2,P＜0.05 ]明显多于Con组[(840±76)个/mm2],而P50D组[(225±66)个/mm2,P＜0.05]明显减少.水迷宫试验中,P50D组空间探索的潜伏期[(42.68±6.18)s,P＜0.05]明显长于Con组[(15.12±3.43)s],P50D组目标象限停留时间[(32.18±5.38)s,P＜0.05]明显少于Con组[(55.66±8.57)s],而P50组、P10组与Con组的差异均无统计学意义.结论 大剂量异丙酚可以抑制新生大鼠的空间学习、记忆能力发育,并可抑制内源性神经干细胞增殖;小剂量的异丙酚反而可以促进内源性神经干细胞增殖.

Abstract：

Objective To investigate the effects of propofol on the development of spatial learning and memory and neuron proliferation of neonatal rats at different doses. Methods 60 neonatal rats were divided into four groups among per litter by using a randomized block design. Three different doses of propofol group were induced with propofol 10 mg/kg( group P10) ,50 mg/kg( group P50) or 50 mg/kg twice( group P50D) by subcutaneous injection respectively. Neuron proliferation at dentate gyrus was detected by using BrdU marker 3 days later.Morris water maze test was carried out on postnatal day 28. Escape latency,time in probe quadrant were recorded.Results Compared to the control group,neuron marked with BrdU at dentate gyrus in group P50D was significantly decreased( (840±76) vs (225 ±66), P＜0.05) ,group P10 was significantly increased( (840 ±76) vs ( 1225± 154), P＜0.05). Compared to the control group,latency of group P50D was significantly increased( ( 15.12 ±3.43 ) s vs (42.68 ± 6. 18 ) s, P ＜ 0. 05 ), time in probe quadrant of group P50D were significantly decreased ( ( 55.66 ± 8.57 ) s vs (32. 18 ± 5. 38 ) s, P＜ 0. 05 ). Compared to the control group, there was no significant difference between group P50 and group P10. Conclusion Propofol given to seven-day-old rats with 50 mg/kg twice by subcutaneous injection suppresses neuron proliferation and impairs development of memory and learning in neonatal rats,but propofol given with 10 mg/kg once promotes neuron proliferation.     

Role of NF-κB in protection of EPO pretreatment on neonatal rat cardiac myocytes with hypoxia/reoxygenation injury

Objective：To observe the protective effects of erythropoietin （EPO） pretreatment on cardiac myocyte with hypoxia/reoxygenation （H/R） injury and the role of NF-κBin this effects. Methods：After the H/R model of cardiac myocytes of neonatal rats was established, the cultured cardiac myocytes were divided into 4 groups, including EPO pretreatment group （ EPO 10 U/ml 24 h before H/R）, EPO pretreatment ＋ PDTC group（EPO 10 U/ml and PDTC 5 μg/ml 24 h before H/R）, PDTC group （PDTC 5 μg /ml 24 h before H/R） and eomrolgroup. Before and after the H/R, assay of LDH concentration in the culture medium, the survival rate of the myocytes tested by MTT chromatometry and the apoptosis by flow cytometry were undertaken. Activation of NF-κB was determined by EMSA before and after H/R. Results：EPO pretreatment markedly reduced the LDH concentration in the medium, elevated the survival rate of myocytes and inhibited the apoptosis after H/R. Addition of PDTC during the pretreatment abol- ished the protective effects of EPO pretreatment. NF-κB was markedly activated during EPO pretreatment and PDTCinhibited the activation. However, after H/R, the activity of NF-κB in myocytes with EPO pretreatment was significantly inhibited compared to the other myocytes. Conclusion：NF-κB is significantly activated during EPO pretreatment, but is inhibited after H/R, which is correlated with the protective effects of EPO pretreatment on cardiac myocytes with H/R. This phenomenon can be explained as the negative feedback mechanism of the activation of NF-κB.     

The Effects of PDTC on Interleukin-1β-induced Nitric Oxide Production in Chondrocytes

In order to find new drugs to inhibit nitric oxide （NO） production, the effects of pyrrolidine dithiocarbamate （PDTC）, a nuclear factor-kappa B （NF-κB） inhibitor, on recombinant human interleukin-1β （rhIL-1β）-induced NO production in chondrocytes were investigated. Rat chondrocytes were isolated and cultured, divided into control, P0, P1, P2, P3 and P4 groups. The chondrocytes in the P0, P1, P2, P3 and P4 groups were treated with different concentrations of PDTC （0, 3, 10, 30, and 50 p.mol/L respectively） for 1 h and then incubated with 5 U/mL rhIL-1β for 24 h. NO assay kit and RT-PCR were used to detect the NO content and the iNOS mRNA expression in the chondrocytes The expression level of iNOS mRNA in control, P0, P1, P2, P3 and P4 groups was 0.02±0.01, 1.24±0.13, 1.21±0.14, 0.61±0.11, 0.40±0.09, 0.21±0.06, and the relative content of NO was 15.8±2.7, 100±14.8, 92.6±9.3, 68.3±14.2, 27.5±9.8, 19.8±3.6, respectively. In the P0, P1, P2, P3 and P4 groups, the expression of iNOS mRNA and NO production were significantly increased as compared with those in the control group. As compared with the P0 group, the expression of iNOS mRNA and NO content in control group were lower. In the P2, P3 and P4 groups, PDTC could significantly inhibit the expression of iNOS and NO production induced by rhIL-1β in a concentration-dependent manner. It is suggested that PDTC can inhibit NO production and iNOS mRNA expression induced by IL-1β, which may provide an alternative method for the treatment of osteoarthritis.     

鞘内注射代谢性谷氨酸受体亚型5拮抗剂对骨癌痛小鼠痛行为及脊髓水平星形胶质细胞活化的影响

目的 探讨代谢性谷氨酸受体亚型5(mGluR5)拈抗剂MTEP的抗癌痛作用及其机制.方法 C3H/HeNCrlVr雄性小鼠60只,随机分为:(1)正常组:正常饲养21d;(2)MTEP+Tumor组:癌痛组从第14d开始鞘内注射MTEP150nmol,1次/d,共7次;(3)生理盐水+Tumor组:以等体积的生理盐水代替MTEP;(4)MTEP+Sham组:用等剂量MTEP给予假手术组;(5)生理盐水+Sham组:用等体积的生理盐水给予假手术组.每组12只小鼠.用热辐射刺激仪测定缩足潜伏期(PWTL).各组小鼠均在第21d行为学测试后取脊髓标本,分别应用Real-time PCR和western blot检测胶质细胞纤维酸性蛋白(GFAP)mRNA及蛋白的表达.结果 各组小鼠基础PWTL差异无统计学意义(P＞0.05).在术后14d,与正常组相比,假手术组PWTL差异无统计学意义(P＞0.05),而癌痛组PWTL明显降低(P＜0.05).在术后21 d,和正常组相比,MTEP+Sham组、生理盐水+ShaT组PWTL、脊髓GFAP表达差异均无统计学意义(P＞0.05);生理盐水+Tumor组PWTL[(6.18±1.29)s]明显低于正常组[(15.91±1.65)s]、生理盐水+Sham组[(16.57±1.86)s]和MTEP+Sham组[(17.05±2.43)s](P＜0.05),脊髓水平GFAP表达则高于上述3组;MTEP+Tumor组PWTL[(9.39±1.94)s]高于生理盐水+Tumor组,脊髓水平GFAP表达则低于生理盐水+Tumor组(P＜0.05).结论 鞘内注射MTEP具有抗癌痛作用,抑制脊髓水平星形胶质细胞活化可能是其机制之一.

Abstract：

Objective To investigate effects of metabotropic glutamate receptor subtype 5 (mGluR5) antagonist MTEP on the nociceptive behavior and the expression of glial fibrillary acidic protein (GFAP) in spinal cord associated with bone cancer pain. Methods C3H/HeNCrlVr 60 male mice were randomly divided into 5 groups: ( 1 ) normal control group: the mice were given food and water ad libitum; ( 2 ) MTEP + Tumor group: the mice were treated by intrathecal gdministration ( once daily on the days 14 ～20 after inoculation of tumor cells)with MTEP (150 nmol); (3) physiological saline + Tumor group:the tumor mice were treated with the same volume of physiological saline; (4) MTEP + Sham group: the sham mice were treated with the same dose of MTEP;(5) physiological saline + Sham group: the sham mice were treated with the same volume of physiological saline.the mice pain behaviors were assessed with the paw withdrawal thermal latency (PWTL) at the corresponding time points, then the mice were killed and the samples of spinal cord were used to real-time PCR and western blot detection of GFAP mRNA and protein expression. Results The basic values of PWTL had no significant differences among all groups (P＜0.05). At day 14 after operation,no significant difference was found in the PWTL value between normal control group and the sham operation group. But in tumor group, the PWTL value was significantly lower than in the normal control group (P＜ 0.05 ). At day 21 after operation,the PWTL and the level of GFAP expression in the spinal cord had no significant differences among normal control group, MTEP + Sham group and physiological saline + Sham group (P ＞ 0.05 ); the PWTL ( (6. 18 ± 1.29 ) s) in physiological saline + Tumor group was significantly lower than in normal control group ( ( 15.91 ± 1.65 )s), physiological saline + Sham group ( ( 16.57 ± 1.86) s) and MTEP + Sham group ( ( 17.05 ± 2.43 ) s) (P ＜ 0.05 ), but the level of GFAP expression was higher than in the above three groups. In MTEP +Tumor group ,the PWTL (9.39 ± 1.94s) was higher than in physiological saline + Tumor group, and the level of GFAP expression was lower than in physiological saline +Tumor group (P ＜ 0.05 ). Conclusion Inhibiting spinal activation of astrocytes may be one of the MTEP anticancer pain mechanisms.     

The influence of insulin on secretion of IGF-Ⅰ and IGFBP-Ⅰ in cultures of human endometrial stromal cells

Objectives To study the influence of insulin on IGF-Ⅰ and IGFBP-Ⅰ secretion of the human endometrial stromal cells. Methods Late proliferative phase endometrial stromal cells were isolated from endometrium tissues and then cultured for 24 h in Hams F-12 only as a control and in Hams F-12 with different concentrations of estradiol (E2) and insulin (INS) as treated groups. Simultaneously, the endometrial stromal cells from late secretory phase endometrium were cultured for 24 h in Hams F-12 only as a control and in Hams F-12 supplemented with different concentrations of progesterone (P) and insulin as treated groups. After 24 h of culturing, the mediums were collected for either IGF-Ⅰ or IGFBP-Ⅰ assays. Result The concentrations of IGF-Ⅰ in medium from cultured endometrial stromal cells in the proliferative phase were 0.78±0.47 ng/ml in the hormone-free control group; 1.44±0.59 ng/ml and 1.39± 0.33 ng/ml in 100 pg/ml E2 group and 20 μU/ml INS group, which was higher than that of the control group (P<0.05 and P<0.01, respectively). The IGF-Ⅰ concentration in the 100 μU/ml INS group was 2.03±0.53 ng/ml, which was higher than that of the 20 μU/ml INS group (P<0.01). Levels of IGF-Ⅰ in the 100 pg/ml E2 plus 20 μU/ml INS group was 2.18±0.36 ng/ml, which was significantly higher than that of the 20 μU/ml INS and 100 pg/ml E2 group (P<0.01), but lower than that of the 100 pg/ml E2 plus 100 μU/ml INS group (3.42±0.75 ng/ml), P<0.01. The concentration of IGFBP-Ⅰ in medium from cultured endometrial stromal cells in the secretory phase was 2.50±1.39 ng/ml in the hormone-free control group and 5.44±2.09 ng/ml in the 10 pg/ml P group, which was significantly higher than that of the control (P<0.01). IGFBP-Ⅰ concentration in 20 μU/ml INS group was 0.16±0.58 ng/ml, which was lower compared with control, but higher compared with the 100 μU/ml INS group (P<0.01). The level of IGFBP-Ⅰ in the 10 ng/ml P plus 20 μU/ml INS group was 2.10±1.17 ng/ml, lower compared with the 10 ng/ml P group, but higher compared with the 10 pg/ml P plus 100 μU/ml INS group, P<0.01. Conclusions Insulin can stimulate basal (without hormone) and E2-stimulated IGF-Ⅰ secretion in cultured stromal cells from human late proliferative endometrium in a dose-dependent manner. Insulin can suppress basal (without hormone) and P-stimulated IGFBP-Ⅰ secretions in cultured stromal cells from human secretory endometrium in a dose-dependent manner.     

Remote postconditioning induced by brief pulmonary ischemia and reperfusion attenuates myocardial reperfusion injury in rabbits

Background The lung is one of the most important organs that are sensitive to ischemia. We hypothesized that remote postconditioning (RPostC) induced by brief occlusion and reperfusion of the pulmonary artery could attenuate myocardial reperfusion injury.Methods Thirty rabbits were randomized into three groups. Group ischemia-reperfusion (IR) (n=10) were anesthetized rabbits subjected to 30-minute occlusion of the left anterior descending coronary artery followed by 180-minute reperfusion. Group RPostC (n=10) had the left pulmonary artery blocked for five minutes followed by a 5-minute reperfusion, and the left anterior descending coronary artery (LAD) occluded for 30 minutes with a 180-minute reperfusion. Group L-Nw-nitro-L-arginine methylester (L-NAME) + RPostC (n=10) had the left pulmonary artery blocked for five minutes followed by a 5-minute reperfusion and intravenous infusion of L-NAME (10 mg/kg), and the LAD occluded for 30 minutes with a 180-minute reperfusion. Blood samples were taken for levels of creatine kinase (CK),superoxide dismutase (SOD) and malondialdehyde (MDA) at three different time points. At the end of the experiment,tissue samples of the infarcted region were harvested to calculate the cardiomyocyte apoptosis index (Al) by TUNEL. A piece of left and right lung tissue was harvested to evaluate the damage to the lung.Results After reperfusion for 180 minutes, the concentration of CK was lower in group RPostC, (4.79±0.27) U/ml, than that in group IR, (6.23±0.55) U/ml (P ＜0.01), and group L-NAME + RPsotC, (5.86±0.42) U/ml (P ＜0.01). The concentration of MDA was lower in group RPostC, (6.06±0.36) nmol/ml, than that in group IR, (11.41±0.91) nmol/ml (P ＜0.01), and group L-NAME + RPostC, (11.06±0.62) nmol/ml (P＜0.01). The activity of SOD was higher in group RPostC,(242.34±25.02) U/ml, than that in group IR, (148.05±18.24) U/ml (P＜0.01), and group L-NAME + RPostC, (160.66±9.55) U/ml (P＜0.01). The apoptosis index was lower in group RPostC, (14.25±5.20)%, than that in group IR, (35.77±10.09)% (P ＜0.01), and group L-NAME + RPostC, (30.37±7.76)% (P ＜0.01). No significant difference caused by pulmonary ischemia was found in the lung tissue among the three groups.Conclusions RPostC may attenuate myocardial ischemia-reperfusion injury connected to the activity of endothelial nitric oxide synthase. Brief pulmonary ischemia may not be harmful to lungs.     

电针对坐骨神经分支选择性损伤大鼠痛觉过敏及脊髓NOS活性、NO水平的影响

目的 探讨电针对神经病理性痛大鼠痛觉过敏以及脊髓一氧化氮合酶(NOS)活性和一氧化氮(NO)水平的影响.方法 40只SD大鼠,随机分为空白组、假手术组、手术组和电针组(n=10).采用坐骨神经分支选择性损伤模型,电针"委中"和"环跳"穴,观察其对大鼠机械痛阈和热痛阈的影响,以分光光度法测定脊髓总NOS以及nNOS、iNOS活性和NO水平.结果 坐骨神经分支选择性损伤术可明显降低大鼠机械痛阈,手术组10d时为(14.83±0.79)g,与空白组和假手术组相比,差异有显著性(P＜0.05);并且手术组脊髓总NOS和nNOS活性以及NO水平均明显升高,分别为(44.23±7.81)U/mgprot和(39.64±10.28)U/mgprot以及(112.79±18.01)μmol/g,与空白组和假手术组相比,均差异有显著性(P＜0.01),iNOS却有降低的趋势;而电针干预后大鼠脊髓总NOS和nNOS活性明显降低,分别为(32.14±12.05)U/mgprot和(20.97±12.16)U/mgprot,NO合成也明显减少,为(70.96±18.15)μmoL/g,与手术组各项比较差异有显著性(P＜0.05,P＜0.01);与此同时显著减轻坐骨神经分支选择性损伤大鼠的机械痛敏状态,进而改善其痛行为.结论 电针减轻神经病理性痛可能与其抑制脊髓NOS活性、降低NO水平有关.     

褪黑素对噪声应激大鼠学习记忆的保护作用

目的 探讨噪声应激对大鼠记忆功能的影响及可能机制.方法 将50只大鼠用随机数字法分为空白组和2个实验组、2个对照组.空白组不给任何处理,实验组及对照组暴露于120dB噪声应激1d或3d,8h/d,并于每次噪声应激前分别腹腔注射褪黑素(MT,15mg/kg)或等量生理盐水.应激结束后,用旷场反应箱测试大鼠的行为,用Morris水迷宫法测试大鼠学习记忆,用试剂盒检测大鼠大脑皮层和海马中一氧化氮(NO)、超氧化物歧化酶(SOD)和丙二醛(MDA)含量.结果 无论噪声应激1d或3d,旷场反应测试大鼠兴奋性和探索性行为实验组[运动总距离(TMD)(1322.50±504.32)cm,(1819.55±458.37)cm,较快运动时间(FMT)(68.49±23.90)s,(87.34±16.01)s,距中心距离(DTC)(63.56±2.75)cm,(60.13±1.87)cm,内环内时间(ITT)(7.87±2.06)s,(9.60±2.89)]均明显小于对照组[TMD(2042.03±449.19)cm,(2325.73±384.90)cm,FMT(109.32±21.84)s,(124.65±16.74)s,DTC(58.00±1.53)cm,(55.05±5.13)cm,ITT(12.84±3.62)s,(14.92±2.75)s,(P＜0.05,P＜0.01)];水迷宫中的逃避潜伏期实验组大鼠[(10.69±3.37)s,(18.87±4.74)s]均明显小于对照组[(23.86±7.66)s,(33.55±7.20)s,(P＜0.01)];大鼠大脑皮层和海马组织中NO和MDA含量实验组[皮层NO(3.35±0.40)μmol/gprot,(4.50±0.41)μmol/gprot,海马NO(2.24±0.18)μmol/gprot,(3.15±0.21)μmol/gprot,皮层MDA(1.34±0.44)nmol/mgprot,(2.39±0.18)nmol/mgprot,海马MDA(0.13±0.07)nmol/mgprot,(0.53±0.10)nmol/mgprot]明显低于对照组[皮层NO(3.35±0.40)μmol/gprot,(5.03±0.44)μmol/gprot,海马NO(2.93±0.31)μmol/gprot,(3.38±0.24)μmol/gprot,皮层MDA(2.24±0.26)nmol/mgprot,(4.21±0.21)nmol/mgprot,海马MDA(0.47e0.29)nmol/mgprot,(1.33±0.187)nmol/mgprot,(P＜0.05,P＜0.01)],SOD含量实验组[皮层(763.95±214.36)U/mgprot,(491.33±35.85)U/mgprot,海马(817.02±232.39)U/mgprot,(644.85±28.02)U/mgprot]明显高于对照组[皮层(556.50±101.51)U/mgprot,(327.35±30.54)U/mgprot,海马(279.74±117.02)U/mgprot,(108.75±15.52)U/mgprot,(P＜0.05,P＜0.01)].结论 褪黑素对噪声应激大鼠记忆障碍有改善作用,这种作用可能与抑制噪声应激大鼠大脑皮层和海马中NO及MDA的升高,并提高SOD活性作用有关.     

埃索美拉唑对大鼠创伤性脑损伤后肠黏膜屏障损伤的作用

目的 探讨埃索美拉唑对大鼠创伤性脑损伤(TBI)后应激性肠黏膜损伤的保护作用.方法 雄性Wistar大鼠54只,随机分为3组,各18只:TBI组埃索美拉唑预处理组[质子泵抑制(PPI)组],假手术组(对照组).TBI组动物制模前1 h皮下注射埃索美拉唑0.1 mg(0.25 mi/100 g),TBI组和对照组术前1 h注射等量的生理盐水(0.25 ml/100 g).各组动物分别在术后3、12、24 h心脏取血后处死(各时间点6只),处死后取脑组织和肠黏膜组织观察形态学改变,测定肠黏膜组织二胺氧化酶(DAO)、超氧化物歧化酶(SOD)、丙二醛、羟自由基、髓过氧化物酶(MPO)的含量,同时检测血清中DAO活性及异硫氰酸荧光素(FITC)浓度.结果 (1)PPI组肠黏膜损伤程度较TBI组轻.(2)TBI组损伤后随时间延长血清FITC-右旋糖苷外渗逐渐增加,以24 h时间点最为明显(P＜0.01),PPI组较TBI组轻[(3720±401)ng/ml比(5230±489)ng/ml P＜0.05].(3)肠黏膜组织中DAO的活性随时间增加逐渐下降,以24 h点最为明显,PPI组下降幅度高于TBI组,血清变化与之相反.(4)TBI组肠黏膜组织中SOD、GSH随时间延长逐渐下降,以24 h点最为明显(P＜0.05),PPI组SOD和GSH分别高于TBI组(U/mgprot:13.0±2.4和208±48比10.2±2.8和140±46,均P＜0.05).(5)TBI组肠黏膜组织中羟自由基、丙二醛、MPO随时间延长逐渐升高,以24 h点最为明显(P＜0.05),PPI组羟自由基、丙二醛和MPO均低于TBI组(U/mgprot:108±8、6.2±0.6、1.53±0.52比150±8、7.7±0.9、1.93±0.53,均P＜0.05).结论 创伤性脑损伤可能导致应激性肠黏膜损伤,氧自由基在致肠黏膜损伤中起重要作用,埃索美拉唑通过抗氧化和抑制中性粒细胞活性可以减轻应激性肠黏膜损伤.     

酸枣仁汤对老年失眠证候模型大鼠学习记忆及脑内自由基、一氧化氮合酶的影响

目的 探讨酸枣仁汤对失眠证候模型大鼠学习记忆及脑组织丙二醛(MDA)含量及超氧化物歧化酶(SOD)、一氧化氮合酶(NOS)活性的影响.方法 用D-半乳糖致亚急性衰老、环磷酰胺及氧化可的松致阴血亏虚,及自制改良多平台法剥夺睡眠,制作老年阴血亏虚型失眠证候大鼠复合模型,观察大鼠学习记忆、大脑皮质及海马部位MDA含量及SOD、NOS活性的变化,以及酸枣仁汤对这些指标的影响.结果 与环境对照组[(47.3 ±4.6)s,(9±1.4)s,(6.5±1.2)次]比较,证候模型组[(108.9±12.5)s,(89±11.5)s,(0)次]睡眠剥夺48 h后,大鼠学习记忆能力显著下降(t=4.36,3.18,2.07,P＜0.01),证候模型组皮质、海马部位MDA含量和SOD、NOS活性[(3.8±0.6)、(3.0±0.5)nmol·mgprot-1,(229.7±25.8)、(236.3±25.2)U·mgprot-1,(5.7±0.8)、(5.4 ±0.9)U·mgprot-1]相对于环境对照组[(2.1±0.4)、(1.6±0.4)nmol·mgprot-1,(155.5±10.6)、(147.2±26.1)U·mgprot-1,(2.8±0.7)、(2.9±0.5)U·mgprot-1]均升高(t=2.89,3.01,6.78,5.94,3.10,3.46,P＜0.05);经酸枣仁汤高、低剂量干预后大鼠学习记忆能力得以明显改善(P＜0.05),高剂量组皮质、海马部位MDA含量和SOD、NOS活性明显下降(P＜0.05),低剂量组也明显下降(P＜0.05),并趋于正常范围.结论 酸枣仁汤对老年失眠证候模型大鼠的学习记忆能力障碍有改善作用,其机理之一可能与减轻睡眠剥夺中自由基和NO对神经细胞的损伤有关.     

银杏叶、蓝莓提取物与复合营养素对老龄大鼠学习记忆及抗氧化功能的影响

目的 观察银杏叶、蓝莓提取物与复合营养素对老龄大鼠学习记忆功能及抗氧化能力的影响.方法 15月龄Wistar大鼠20只,随机分为对照组和干预组,每组10只,雌雄各半.对照组和干预组大鼠分别饲喂基础饲料及添加复合营养素的自制饲料,同时灌胃蒸馏水或植物提取物混合液.灌胃剂量为4ml·kg-1·d-1,实验期10周,干预结束后采用水迷宫实验检测大鼠的学习记忆功能.然后处死动物.荧光法测定血清维生素E含量;紫外可见分光光度法测定血清总抗氧化能力(T-AOC)、抑制羟自由基能力;全血谷胱甘肽过氧化物酶(GSH-Px)活性采用DTNB法测定.结果 干预组老龄大鼠水迷宫实验训练期及检测期逃避潜伏期[分别为(7.74±1.84)s,(6.85±0.82)s]均低于对照组[分别为(10.14±2.05)s,(9.92±2.30)s],差异具有显著性(t=4.506,t=4.300,P＜0.05).干预组老龄大鼠血清VitE含量[(16.22 ±5.03)μg/ml]高于对照组[(7.52±4.24)μg/ml],差异具有显著性(t=6.058,P＜0.05).干预组老龄大鼠血清T-AOC、GSH-Px活性[分别为(13.01±9.36)U/ml,(47.27±8.60)活力单位]高于对照组[(6.51±1.39)U/ml,(33.48 ±3.19)活力单位],差异具有显著性(t=4.113,t=7.679,P＜0.05).与对照组比较,干预组老龄大鼠的学习记忆功能得到改善;血清维生素E含量增加;T-AOC、抑制羟自由基能力提高;GSH-Px活性升高.结论 银杏叶、蓝莓提取物与复合营养素对老龄大鼠的学习记忆功能有改善作用,其作用途径可能与提高抗氧化防御系统功能有关.     

阻塞性睡眠呼吸暂停低通气综合征患者心脏结构和功能的观察

目的 探讨阻塞性睡眠呼吸暂停低通气综合征(OSAHS)对患者心脏结构和功能的影响.方法 OSAHS患者75例,根据病情分为轻度(25例)、中度(25例)及重度组(25例);根据病程分为＜5年(22例)、5-10年(26例)和＞10年(27例)组;健康人25名,为对照组.对各组受试者进行超声心动图检查,记录主动脉内径、肺动脉内径、心腔大小、室间隔厚度、舒张末期右心室前壁厚度及运动幅度、左心室射血分数(LVEF)、短轴缩短率(FS)、E峰/A峰(E/A))比值等心脏参数并进行比较.结果 轻、中、重度组和对照组肺动脉内径分别为(21.4±2.5)、(24.7±2.0)、(26.7±2.1)、(21.2±2.7)mm,右心室内径分别为(19.0±1.8)、(22.0±2.1)、(23.9±2.1)、(18.9±1.8)mm,右心室前壁厚度分别为(4.7±1.2)、(6.5±1.3)、(7.5±1.4)、(4.1±1.0)mm,中、重度组均明显大于对照组和轻度组(均P＜0.01),重度组明显大于中度组(均P＜0.01);LVEF分别为(63.1 ±8.1)%、(60.0 ±10.2)%、(54.5±9.1)%、(63.6±7.7)%,FS分别为(38.1±4.3)%、(37.0 ±6.4)%、(33.6 ±5.4)%、(39.5±4.9)%,E/A比值分别为(1.13±0.13)、(0.96±0.16)、(0.85±0.12)、(1.28 ±0.15),重度组LVEF和FS均明显低于对照组和轻、中度组(均P＜0.05),轻、中、重度组E/A比值均明显低于对照组(P＜0.01),轻、中、重度组间两两比较差异均有统计学意义(均P＜0.01).＞10年组右室内径、肺动脉内径和右室前壁厚度均明显大于5～10年组(P＜0.05,P＜0.01),5-10年组明显大于＜5年组和对照组(P＜0.05,P＜0.01);＞10年组LVEF、FS和E/A比值均明显低于5-10年组、＜5年组和对照组(P＜0.05,P＜0.01).结论 OSAHS可导致患者心脏结构和功能均发生改变,并随着OSAHS病情的加重和病程的延长而更加明显.     

丹参多酚酸对大鼠缺血心肌线粒体酶的影响

目的:探讨丹参多酚酸对大鼠心肌缺血再灌注过程中线粒体Na+,K+-ATP酶和Ca2+-ATP酶的影响。方法:采用健康Wistar大鼠(n=40)建立Langendorff离体心脏灌注模型,并分为4组。对照组用Krebs液持续灌注120 min;缺血再灌注组平衡30 min后全心缺血30 min,再灌注60 min;丹参多酚酸预处理组平衡5 min后加入丹参多酚酸使其在灌流液中终浓度为14μmol/L,持续灌注25 min后全心缺血30 min,再灌注60 min;丹参多酚酸后处理组平衡30 min后全心缺血30 min,加入丹参多酚酸使其在灌流液中终浓度为14μmol/L,再灌注60 min。TTC染色观察心肌梗死面积,并用心肌梗死面积、乳酸脱氢酶(LDH)活性和心脏血流动力学(包括心率、主动脉流量、冠状动脉流量和心输出量)的变化评估大鼠心肌的损伤程度。利用差速离心法提取线粒体,并通过紫外分光光度法测定心肌线粒体Na+,K+-ATP酶和Ca2+-ATP酶的活性。结果:丹参多酚酸预处理组的心肌梗死面积和LDH活性均显著低于缺血再灌注组([24.47±2.14)%比(34.30±2.31)%、(81.46±13.39)U/g比(142.6±6.46)U/g,均P<0.05],缺血再灌注组与后处理组无明显差异。丹参多酚酸对缺血再灌注心肌的血液动力学没有明显作用,只是冠脉流量有较小幅度的增加。丹参多酚酸预处理组线粒体Na+,K+-ATP酶和Ca2+-ATP酶活性显著高于缺血再灌注组([6.12±0.42)U/mg比(4.29±0.28)U/mg、(3.42±0.16)U/mg比(2.62±0.96)U/mg,均P<0.05],而与对照组无明显差异。丹参多酚酸后处理组线粒体Na+,K+-ATP酶和Ca2+-ATP酶活性与缺血再灌注组无统计学差异。结论:丹参多酚酸可以维持心肌线粒体Na+,K+-ATP酶和Ca2+-ATP酶的活性,通过降低缺血再灌注对心肌线粒体的损伤保护心肌。     

趋化因子配体2中和抗体对骨癌痛大鼠痛行为及脊髓小胶质细胞活化的影响

目的 探讨鞘内注射趋化因子配体2(CCL2)中和抗体对胫骨癌痛大鼠机械痛行为及脊髓小胶质细胞活化的影响.方法 采用Walker 256乳腺癌细胞,构建大鼠胫骨癌痛模型.雌性SD大鼠(160～180 g)40只,随机分为5组(n=8):假手术组(Ⅰ组)、假手术+CCL2中和抗体组(Ⅱ组)、骨癌痛组(Ⅲ组)、骨癌痛+ control IgG组(Ⅳ组)和骨癌痛+CCL2中和抗体组(Ⅴ组).术后第10～12天,每天1次鞘内注射CCL2中和抗体或control IgG( 10 μg/15μ1).分别于术前、术后1d,3d,5d,7d,10 d,14 d,21 d测定各组大鼠机械痛敏阈值(PMWT).另取30只大鼠,分组同前(n=6),术后14d取材,免疫荧光染色法测定各组大鼠脊髓背角小胶质细胞标志物(Iba-1)的平均光密度值(MOD).结果 (1)痛行为学:术后10d,14 d,21 d,Ⅲ组大鼠PMWT分别为:(1.78 ±0.38)g,( 1.70±0.17)g,(1.35±0.07)g;Ⅳ组大鼠PMWT分别为:(2.99±0.67)g,(2.52±0.75)g,(1.13±0.07)g;Ⅴ组大鼠PMWT分别为(5.88±0.66)g,(7.81±0.75)g,(6.19±0.53)g.与Ⅲ组相比,Ⅴ组大鼠PMWT值明显升高(P＜0.01),Ⅳ组大鼠PMWT值无明显变化(P＞0.05).(2)免疫荧光染色:术后14d,Ⅲ、Ⅳ、Ⅴ组大鼠脊髓背角Iba-1的MOD值分别为(151.3±10.8),(149.2±10.6),(74.5±5.0);与Ⅲ组相比,Ⅴ组大鼠MOD值明显升高(P＜0.01);Ⅳ组大鼠MOD值无明显变化(P＞0.05).结论 鞘内注射CCL2中和抗体能显著减轻胫骨癌痛大鼠的机械痛敏,并能抑制其脊髓小胶质细胞的活化.     

雄附散抑制大鼠肺间质纤维化过程中肺组织iNOS的表达水平

目的探讨诱导型一氧化氮合酶（iNOS）在雄附散防治大鼠肺间质纤维化机制中的可能作用。方法将70只SD大鼠随机分为正常组,假手术组,模型组,醋酸泼尼松组（5.6 mg/kg）,雄附散高、中、低剂量组（1.4 g/kg,0.7 g/kg,0.35 g/kg）。各组大鼠于造模后第2天开始通过灌胃干预4周（生理盐水0.014 L/kg或相应药物0.014 L/kg）,28 d后取右中肺组织进行组织学观察和iNOS活性测定。结果组织学变化支持雄附散高剂量组可以有效阻止模型大鼠肺组织的纤维化;高、中剂量组肺组织中iNOS酶活性（12.00±3.74）U/mgprot和（13.08±5.14）U/mgprot明显低于其它各干预组（P〈0.01）。结论有效抑制肺组织中iNOS酶活性可能是雄附散有效对抗肺组织纤维化机制中的重要环节。     

鞘内注射促肾上腺皮质激素释放因子对肠易激综合征模型大鼠内脏高敏感性的影响

目的 研究鞘内注射促肾上腺皮质激素释放因子(CRF)对肠易激综合征模型大鼠内脏高敏感性的影响.方法 将30只大鼠随机分成3组,每组各10只,应用化学刺激方法建立肠易激综合征大鼠模型后,分别给予鞘内注射CRF或预先腹腔注射CRF-1受体拮抗剂CP-154526后再鞘内注射CRF,对照组则注射生理盐水;实验后所有大鼠给予直肠内球囊扩张,检测球囊扩张引起腹部收缩反射的最小容量阈值及球囊不同容量扩张时腹部收缩反射的次数,以评价鞘内注射CRF对肠道敏感性的影响.应用免疫组织化学染色及计算机图像分析系统半定量分析脊髓后角即刻早基因(c-fos)表达.结果 直肠内球囊扩张时,鞘内注射CRF组引起腹部收缩的最小容量阈值为(0.62±0.10)ml,高于其他2组[分别为(0.52±0.09)ml、(0.56±0.08)ml;F=3.25;P＜0.05].直肠球囊扩张体积1.0 ml时鞘内注射CRF组腹部收缩反射次数为(9.10±1.97)次,明显低于其他2组[分别为(14.4±1.71)次、(15.6±2.32)次;F=29.4;P＜0.01];体积1.5 ml、2.0 ml高容量扩张时3组间差异无显著性(P＞0.05).鞘内注射CRF组腰骶段脊髓后角c-fos阳性神经元细胞相对切面面积及OD值均明显低于其他2组(P＜0.01).结论 鞘内注射CRF能够降低肠易激综合征模型大鼠内脏高敏感性,并减少其腰骶段脊髓后角c-fos的表达.