严重创伤早期大鼠下丘脑HSP70与iNOS表达与分布的时效关系

目的研究严重创伤早期大鼠下丘脑热休克蛋白(HSP)70与诱导型一氧化氮合酶(iNOS)转录表达变化分布和意义.方法采用BIM-Ⅲ型生物撞击机致大鼠胸部严重撞击伤,并造成单侧股骨骨折,运用S-ABC免疫组化、原位杂交技术测定下丘脑HSP70与iNOS及其mRNA的转录表达水平.结果正常对照组HSP70低水平表达,在伤后1小时即开始明显增高(P＜0.05),6小时达到峰值为正常的18倍,12小时后开始下降,24小时时主要集中在室旁核内;iNOS无基础表达,在伤后1小时开始出现,随HSP70同步增高,8小时达到高峰,较1小时时增加13倍.两者相关系数r=0.97601.结论严重创伤后早期下丘脑内HSP70和iNOS过度表达, 在严重创伤应激时下丘脑的损伤与抗损伤机制中起重要作用.     

失血性休克及复苏后大鼠肠上皮UCP2变化及其线粒体功能损伤的研究

目的研究失血性休克及复苏后解偶联蛋白2(UCP2)在肠上皮线粒体功能损伤中的作用.方法复制大鼠失血性休克复苏模型, 分别于休克前、休克90分钟及复苏2小时、6小时、12小时、24小时取小肠上皮,用Western-blot法测线粒体UCP2的含量,用荧光分光光度法测UCP2对线粒体膜电位(MP)、线粒体内活性氧(ROS)产生的影响作用,用高效液相色谱法(HPLC)测组织中三磷酸腺苷(ATP)的含量.结果 (1)失血性休克及复苏后肠上皮线粒体中UCP2含量增高 (P<0.05);(2)UCP2可降低MP并抑制线粒体内ROS产生,但UCP2对线粒体内ROS产生的调节作用受MP影响;(3)失血性休克后,肠上皮组织中ATP含量明显下降,是对照组的20.81%,复苏后24小时,其水平仍低于正常.结论 UCP2可能参与失血性休克及复苏后肠上皮线粒体功能的损伤.     

慢性前列腺炎患者精浆中热休克蛋白的表达及临床意义

目的　探讨慢性前列腺炎患者精浆中热休克蛋白 70 (HSP 70 )的表达及临床意义。　方法　采用酶联免疫吸附测定法 (ELISA)对 10例慢性细菌性前列腺炎 (Ⅱ型 )、10例炎症型慢性非细菌性前列腺炎即炎症型慢性骨盆疼痛综合征 (ⅢA 型 )、10例非炎症型慢性非细菌性前列腺炎即非炎症型慢性骨盆疼痛综合征 (ⅢB 型 )患者及 6例健康志愿者精浆中HSP 70含量进行测定 ,并与慢性前列腺炎症状评分 (修订版 )进行相关性研究。　结果　Ⅱ型、ⅢA 型、ⅢB 型慢性前列腺炎患者及正常对照组精浆HSP 70含量分别为 (10 .74± 4 .0 2 )ng/ml、(2 .5 8± 0 .96 )ng/ml、(2 .78± 1.12 )ng/ml及 (6 .5 7±2 .72 )ng/ml。Ⅱ型患者精浆HSP 70含量显著高于正常对照组 ,P <0 .0 1,差异有显著性意义。ⅢA 型与ⅢB 型患者HSP 70含量均显著低于正常对照组 ,P <0 .0 1,差异有显著性意义。而ⅢA 型与ⅢB 型患者HSP 70含量间差异无显著性意义 ,P >0 .0 5。各组精浆HSP 70含量与慢性前列腺炎症状评分间均呈显著的负相关关系 (P <0 .0 1)。　结论　测定精浆中HSP 70的表达对各种类型慢性前列腺炎诊断有一定的临床应用价值 ,并可作为评价慢性前列腺炎病情程度的分子生物学指标。HSP 70在慢性前列腺炎的治疗方面也具有广阔的应用前景。     

Dobutamine compensates deleterious hemodynamic and metabolic effects of vasopressin in the splanchnic region in endotoxin shock

BACKGROUND: Vasopressin is a potent vasopressor in septic shock, but it may impair splanchnic perfusion. We compared the effects of vasopressin alone and in combination with dobutamine on systemic and splanchnic circulation and metabolism in porcine endotoxin shock. METHODS: Twelve pigs were randomized to receive either vasopressin (VASO, n = 6) or vasopressin in combination with dobutamine (DOBU, n = 6) during endotoxin shock (E. coli endotoxin infusion). Endotoxin infusion rate was increased to induce hypotension after which vasoactive drugs were started. We aimed to keep systemic mean arterial pressure (MAP) >70 mmHg by vasopressin; the goal of dobutamine infusion was to prevent decrease in cardiac output often associated with vasopressin infusion. Regional blood flows, oxygen delivery and consumption, arterial and regional lactate concentrations were measured. RESULTS: Mean arterial pressure >70 mmHg was achieved in both the VASO and DOBU groups. After the primary decrease of cardiac output by vasopressin, systemic blood flow remained stable in vasopressin-treated animals. However, vasopressin as a monotherapy decreased portal venous blood flow. This was prevented by dobutamine. Vasopressin also induced splanchnic lactate release and arterial hyperlactatemia, which were not observed when dobutamine was combined with vasopressin. CONCLUSION: Dobutamine prevents adverse hemodynamic and metabolic effects of vasopressin in septic shock.     

Ergothioneine pretreatment protects the liver from ischemia-reperfusion injury caused by increasing hepatic heat shock protein 70

BACKGROUND: Reperfusion of the liver after ischemia induces the expression of the heat shock genes and the synthesis of the heat shock proteins (HSP). We studied the effects of the natural antioxidant ergothioneine (EGT) treatment on the expression of HSP70 in ischemic-reperfused (IR) liver. METHODS: Adult male Wistar rats were randomly divided into three groups: Sham group given standard laboratory chow and water for 3 weeks followed by sham operation; Control group given standard laboratory chow and water for 3 weeks followed by liver IR injury; EGT group given standard laboratory chow supplementation l-ergothioneine (1.2 mg/kg/d body weight) administered by gavage and water for 3 weeks followed by liver IR injury. Ten rats from each group were killed to determine serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactic dehydrogenase (LDH), tissue malondialdehyde (MDA), HSP70 levels, and histologic changes at 30, 60, and 120 min of reperfusion, respectively. Survival was followed for 1 week. RESULTS: IR caused significant increase in serum AST, ALT, LDH, and tissue MDA levels. As compared with the control group, animals treated with EGT experienced a significant decrease in serum AST, ALT, and LDH levels in all reperfusion periods. Tissue MDA levels in animals receiving EGT were significantly reduced as compared with control group at 30 min and 60 min after reperfusion. After ischemia, reperfusion caused a remarkable production of HSP70 in the control group. When the rats were pretreated with EGT, the levels of HSP70 increased significantly in their livers after reperfusion compared with the control group. Liver injury in the EGT-treated animals was lower to that in the control group. The 7-day survival rate was significantly improved (from 50% to 80%) by EGT pretreatment. CONCLUSION: HSP70 has been shown to induce tolerance against warm IR injury in rat livers. EGT pretreatment protects the liver from IR injury by over-expression of HSP and the subsequent suppression of lipid peroxidation.     

低能量体外冲击击波碎石术研究进展

复式脉冲低能量体外冲击波碎石术治疗尿路结石成功率高,复打率低,副作用少,无严重并发症。本文就低能量体外冲击波碎石术提高成功率、减轻结石周围组织损伤及预防结石再复发的研究进展作一综述。     

Comparison of Effects of Levobupivacaine and Ropivacaine Infiltration on Cutaneous Wound Healing in a Rat Model

ABSTRACT

Purpose: To compare the effects of fluid resuscitation with lactated Ringer's solution (LR) and saline-based 6% hydroxyethyl starch 130/0.4 (HES) on the inflammatory response and oxidative stress in the small intestine as well as on bacterial translocation to the liver. Methods: Sprague–Dawley rats were subjected to blood pressure-controlled hemorrhagic shock and then resuscitated with LR or HES. At 1, 3, 6, 12, and 24 hr after resuscitation, liver tissues were collected to count the bacterial colonies, and small intestines were harvested to analyze the levels of inflammatory (TNF-α and HO-1) and oxidative stress (MPO) mediators as well as the intestinal injury by immunohistochemistry, colorimetry and hematoxylin &; eosin staining, respectively. Results: The expression level of TNF-α in the LR group was stable from 1 to 6 hr but decreased at 12 hr and then abruptly increased at 24 hr. The expression level of TNF-α in the LR group was significantly lower than that in the HES group, especially during the first 12 hr post-fluid infusion. MPO activity decreased to its lowest level at 3 hr but increased from 6 to 12 hr, with no difference at 24 hr between the two groups. Although a decreasing tendency was observed from 6 hr, HO-1 expression levels remained higher in the LR group than in the HES group at 12 and 24 hr, particularly at 12 hr. During the initial 12 hr, the LR group exhibited significantly lower colony-forming units in the liver tissues than the HES group. Chiu's score in the intestine decreased regardless of which resuscitative fluids were used. Conclusions: During early resuscitation (within 12 hr), LR may be superior to HES in reducing intestinal injuries by suppressing inflammatory and oxidative mediators.     

MK2 plays an important role for the increased vascular permeability that follows thermal injury

We previously reported Rho kinase is involved in vessel hyper-permeability caused by burns. Here we further explore the Rho kinase downstream signaling, it is found that its specific inhibitor Y27632 significantly diminishes the activation of JNK and p38 MAPKs but not ERK that induced by serum from burned rats (burn-serum). JNK activation was found involved in the expression of HUVEC adhesion molecules following thermal injury, although not in the process of stress fiber formation. Inhibition of various MAPKs by specific inhibitors showed that SB203580 (inhibitor of p38), but neither SP600125 (inhibitor of JNK) nor PD98059 (inhibitor of ERK), abolish activation of the p38 downstream kinase MK2. Demonstration of stress fibers by fluorescent-labeled phalloidin showed that inhibition of MK2, either by its specific inhibitor or by dominant negative adeno-viral-carried constructs, significantly reduced burn-serum-induced HUVEC stress-fiber formation, while inhibition of another downstream p38 MAPK kinase, PRAK, had no such effects. Transfection of dominant negative adeno-viral MK2 (Ad-MK2(A)) significantly inhibited thermal injury-induced blood vessel hyper-permeability in rats and, moreover, prolonged the survival of burned rats beyond 72 h following thermal injury. One of the mechanisms behind these phenomena is that Ad-MK2(A) causes a significant depression of burn-serum-induced HSP27-phosphorylation, while the adeno-viral transported dominant negative PRAK (Ad-PRAK(A)) does not block. Although the effect of blockade of MK2 through its adeno-viral approach requires further study and investigation of alternatives to know for sure, we may have found a new pathway behind thermal-injury-induced blood vessel hyper-permeability, namely: Rho kinase > p38 > MK2 > HSP27.     

热休克蛋白72肽结合区对肾小管上皮间质转分化的影响

目的 探讨热休克蛋白72肽结合区在肾小管上皮间质转分化(EMT)过程中的作用和可能机制.方法 应用质粒转染方法分别诱导热休克蛋白72(HSP72)野生型、肽结合区缺失型(HSP72-△PBD)和肽结合区(PBD)的表达.用转化生长因子β1(TGF-β1)刺激大鼠肾小管上皮细胞(NRK-52E)48 h,Western印迹和免疫荧光染色检测细胞E-钙黏蛋白(cadherin),α-平滑肌肌动蛋白(SMA),HSP72和Smad3/磷酸化(p)-Smad3蛋白表达.结果 TGF-β1(10 μg/L)刺激NRK-52E细胞48 h后上调α-SMA和下调E-cadherin蛋白表达水平.Western印迹及细胞免疫荧光显示,过表达HSP72和PBD能明显减轻TGF-β1诱导的NRK-52E细胞E-cadherin蛋白表达下调和α-SMA蛋白表达上调,而过表达HSP72-△PBD不能改变上述蛋白的表达.此外,过表达HSP72和PBD显著抑制Smad3的磷酸化.结论 HSP72抑制Smad3活化和EMT的发生可能与PBD的功能有关.     

TNF-α McAb对急性失血性休克大鼠肝脏的作用

目的探讨肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)在急性失血性休克大鼠肝损伤中的作用及TNF-α单克隆抗体的保护作用。方法将24只雄性SD大鼠随机均分为三组:对照组(A组)、休克+乳酸林格氏液复苏组(B组)和休克+TNF-α单克隆抗体复苏组(C组)。B和C组大鼠通过股动脉放血,制作急性失血性休克动物模型;B组用乳酸林格氏液复苏,C组用含TNF-α单克隆抗体(3mg/kg)的乳酸林格氏液复苏,而A组在同等条件下不进行失血。分别检测各组大鼠血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、TNF-α水平和肝组织中MDA、SOD含量,并在光镜和电镜下观察各组大鼠肝组织的病理变化。结果 B、C组大鼠血清ALT、AST、TNF-α水平和肝组织中MDA含量较A组升高,SOD含量降低;C组ALT(343.63±35.61)U/L、AST(748.75±49.76)U/L、TNF-α(99.38±13.16)pg/mL、丙二醛(26.33±1.30)nmol/mgProt较B组ALT、AST、TNF-α、MDA含量降低,C组肝组织中SOD含量(510.14±47.44)U/mgProt较B组升高;在光镜、电镜下观察,C组肝组织损伤较B组减轻。结论 TNF-α可能是急性失血性休克肝损伤的重要因子之一,使用TNF-αMcAb复苏可以减轻失血性休克时大鼠肝损伤,具有一定的保护作用。