高糖诱导人脐静脉内皮细胞衰老过程中活性氧与二甲基精氨酸二甲胺水解酶-非对称性二甲基精氨酸系统的变化

Objective To investigate the changes in reactive oxygen species (ROS) and dimethylarginine dimethylaminohydrolase-asymmetric dimethylarginine (DDAH-ADMA) system in the process of endothelial cell senescence after exposure to high glucose. Methods The human umbilical vein endothelial cells (HUVECs) were cultured with different concentrations of glucose, e.g. 5. 5 mmol/L (normal level),and high levels as 11. 0, 22. 0 and 33. 0 mmol/L. for 48 hours, respectively. Subsequently, SA-β-gal staining was used to evaluate senescence of cells. Telomerase activity was detected by polymerase chain reactionenzyme linked immunosorbent assay (PCR-ELISA). The intracellular ROS level was measured by flow cytometry. The ADMA concentration and DDAH activity were determined with high-performance liquid chromatography. Results Compared with normal glucose concentration group, after the endothelial cells were treated with high glucose concentration (11. 0 - 33. 0 mmol/L) for 48 hours, the number of SA-β-gal positive cells was increased significantly [(7.00±1. 73)%, (12. 67±2. 03)%, (16. 00±2. 26)% vs. (4. 00±1.33)%, P＞0.05, P＜0.05, P＜0.05] and the telomerase activity was inhibited dramatically [(91. 32±4.01)%, (78. 44±3. 78)%, (56. 04±3. 35)% vs. 100%, all P＜0. 05]. The ROS level (mfi) was increased in all high glucose groups (159. 84±27. 52, 188. 99±18. 77, 244. 56±20. 96 vs. 117.11±18. 76, P＜0. 05 or P＜0. 01). At the same time, the ADMA (μmol/L) production was increased (0. 78±0. 14, 0. 88±0.18,1. 08±0.15 vs. 0. 70±0. 12, P＞0. 05, P＜0. 05, P＜0. 05), and DDAH activity was decreased [(91. 32±4.01)%, (78.44±3.78)%, (56. 04± 3. 35)% vs. 100%, all P＜0.05]. Conclusion High glucose can accelerate endothelial cells senescence in dose-dependent manner and the underlying mechanism may be related to an increased oxidative stress and change in DDAH-ADMA system.     

非对称性二甲基精氨酸水平与缺血性脑卒中的相关性

目的 探讨非对称性二甲基精氨酸(asymmetricaldimethylarginine,ADMA)对缺血性脑卒中诊断及病情判断的临床价值.方法 选取88例缺血性脑卒中患者,采用欧洲卒中量表(ESS)进行神经功能缺损评分分为轻度卒中组(31例)、中度卒中组(39例)和重度卒中组(18例).另选30名年龄、性别构成比与患者组差异无统计学意义的健康志愿者为对照组,分别用高效液相色谱联合质谱法测定血浆中ADMA的水平,分析血浆ADMA水平在对照组和缺血性脑卒中各亚组之间的相关性.结果 对照组、轻度卒中组、中度卒中组和重度卒中组的血浆ADMA水平(±s,μmol/L)分别为:1.0±0.10,2.90±0.30,4.82±0.21及6.61±0.23.与对照组血浆ADMA水平比较,患者组各亚组ADMA水平明显升高,其差异有统计学意义(P<0.05);轻度卒中组与中度和重度卒中组的差异有统计学意义(P<0.05);重度组与中度卒中组的差异有统计学意义(P<0.05),与轻度卒中组的差异有统计学显著性意义(P<0.001).结论 血浆ADMA水平与缺血性脑卒中的发病及病变严重程度相关,检测血浆ADMA水平的变化,对缺血性脑卒中的诊断和病情判断、指导治疗有一定的帮助.     

非对称性二甲基精氨酸与血液透析中血压变化的研究

目的研究血液透析(Hemodialysis,HD)患者血浆非对称性二甲基精氨酸(Asymmetric dimethylarginine,ADMA)与透析中血压变化的关系。方法经生物电阻抗检测干体质量达标且符合入选标准的维持性血液透析(Maintenance Hemodialysis,MHD)患者31名进入研究,根据血液透析过程中血压波动情况分为年龄相匹配的3组:透析中高血压组(n=11)、低血压组(n=12)和血压平稳组(n=8)。用酶联免疫吸附(Enzyme linked immunosorbent assay,ELISA)法检测患者透析前、后血浆ADMA水平,探讨ADMA与透析中血压变化的关系,并进行组间矿物质骨代谢指标、电解质、营养指标、炎性标记物、血脂水平、脉压差和降压治疗等的比较。结果 31例MHD患者透析前血ADMA均值为3.37±1.48μmol/L,透析后降至1.71±0.80μmol/L(P0.001),均显著高于国外正常参考值。透析中低血压组透析前、后血ADMA值(4.38±1.56μmol/L,2.25±0.83μmol/L)均高于透析中高血压组和血压平稳组,差异有统计学意义(2.70±1.18μmol/L,1.32±0.60μmol/L和2.78±0.88μmol/L,1.43±0.56μmol/L;P=0.006和0.006)。透析中高血压组患者透析中的平均脉压差高于透析中低血压组和血压平稳组(62.41±11.57mmHg,48.80±12.88 mmHg和44.56±8.30 mmHg,P=0.004)。高血压组碱性磷酸酶(ALP)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)和高敏C-反应蛋白(high-sensitivity c-reactive protein,Hs-CRP)均高于血压平稳组(P值分别为0.036、0.039、0.046、0.046),低血压组同样指标也高于血压平稳组(P值分别为0.046、0.035、0.040、0.004),上述指标在高血压组和低血压组间差异无统计学意义(P0.05)。结论在干体质量达标的MHD患者中,血ADMA水平显著高于正常,透析过程中的血压波动与内皮功能不良、血管僵硬、微炎症状态等密切相关。     

高糖培养脂多糖刺激下肺脏微血管内皮细胞通透性改变及DDAH/NOS/NO失平衡在其发生机制中的作用

目的 探讨高糖对脂多糖(LPS)刺激下血管内皮细胞损伤的影响及其机制.方法 将人肺脏微血管内皮细胞(PMVEC)分为正常糖组(NG组)、正常糖+LPS刺激组(NGL组)、高糖组(HG组)、高糖+LPS刺激组(HGL组),分别给予含10％小牛血清的正常糖(5.5 mmol/L)或高糖(33 mmol/L)培养5d,加入10 mg/L LPS刺激细胞24h.采用免疫荧光染色观察细胞纤维肌动蛋白(F-actin)的分布及变化;扫描电镜观察细胞膜窗孔数量及孔径变化;细胞迁移实验(Transwell)测定单层内皮细胞的辣根过氧化物酶(HRP)通透性;硝酸盐还原法(Griess法)检测细胞培养上清液中一氧化氮(NO)含量;蛋白质免疫印迹试验(Western blotting)测定细胞二甲基精氨酸-二甲胺水解酶2(DDAH2)、诱生型一氧化氮合酶(iNOS)、内皮型一氧化氮合酶(eNOS)的蛋白表达.结果 与NGL组比较,HGL组PMVEC的F-actin分布排列紊乱,细胞膜窗孔异常增大、增多,细胞单层对HRP通透率增加[(53.62±6.70)％比(23.63±3.92)％,P＜0.01],细胞DDAH2表达(积分A值)减少(0.33±0.08比0.77±0.14,P＜0.01),iNOS表达(积分A值)增加(1.40±0.29比1.04±0.09,P＜0.01),eNOS表达(积分A值)减少(0.67±0.09比0.91±0.17,P＜0.05),上清液NO含量(μmol/L)增多(20.36±2.25比7.99±0.33,P＜0.01).结论 高糖加重LPS刺激下体外培养PMVEC的F-actin分布紊乱、单层细胞通透性增加;NO调节紊乱可能参与了PMVEC损害的发生.     

缬沙坦联合辛伐他汀对早期糖尿病肾病大鼠肾脏的保护作用

目的 探讨降脂药辛伐他汀及血管紧张素Ⅱ受体拮抗剂类药物缬沙坦对早期糖尿病肾病大鼠肾脏的保护作用.方法 用链脲佐菌素(STZ)诱导糖尿病肾病大鼠模型,将SD大鼠随机分成5组,每组10只:正常对照组(C组)、糖尿病肾病组(D组)、缬沙坦组(X组)、辛伐他汀组(Z组)和缬沙坦及辛伐他汀联合组(L组).5周末检测5组大鼠血糖(BG)、糖化血红蛋白(HbA1c)、总胆固醇(TC)、甘油三酯(TG)、血尿素氮(BUN)、肌酐(SCr)、肌酐清除率(Ccr)、尿白蛋白排泄率(UAER)等指标的变化;利用透射电子显微镜观察肾脏足细胞超微病理结构.结果 D组与C组比较,BG[分别为(20.3±3.2)、(6.1±0.4)mmol/L]、HbA1c[分别为(7.18±0.47)%、(3.37±0.15)%]、TC[分别为(2.69±0.35)、(1.28±0.24)mmol/L]、TG[分别为(3.09±0.37)、(1.18±0.25)mmol/L]明显升高(P均＜0.05);D组Ccr[(0.89±0.19)ml/min]较C组[(1.27±0.33)ml/min]和X、Z、L组显著下降(P＜0.05),D组大鼠UAER[(19.87±3.85)μg/24 h]明显高于C组[(3.67±1.01)μg/24 h](P＜0.05),X、Z、L组大鼠UAER显著低于D组(P＜0.05),而L组改善尤其显著(P＜0.05);D组的足细胞足突严重融合,X、Z、L组仅少量足突融合较D组改善,而L组改善尤其显著.结论 缬沙坦及辛伐他汀单用及联合应用,尤其是联合用药对早期糖尿病大鼠肾脏有保护作用.

Abstract：

Objective To explore the protection of valsartan combined with simvastatin on kidney in early diabetic nephropathy rats. Methods Diabetic nephropathy rats model were induced by streptozocin (STZ) ,the experimental rats were randomly divided into 5 groups: control (group C), diabetic nephropathy (group D) ,diabetes treated with valsartan (group X) ,diabetes treated with simvastatin (group Z) ,and diabetes treated with combined valsartan and simvastatin ( group L). Blood glucose (BG), HbA1c, blood cholesterol ( TC), trigalloylglycerol ( TG ), blood ureanitrogen ( BUN ), serum creatinine (SCr) , urinary albumin excretion rate (UAER) were measured, and the podocyte ultrastructure was observed by transmission electronic microscopy. Results The levels of BG, HbA1c,TC,TG and UAER in group D increased significantly compared togroup C(BG:[20.3 ±3.2]mmol/L vs [6.1 -±0. 4]mmol/L;HbA1c:[7.18 ±0.47]% vs [3.37 ±0. 15]% ;TC: [2. 69 ±0. 35] mmol/L vs [1.28 ±0. 24] mmol/L;TG: [3.09 ±0. 37] mmol/L vs [1.18 ±0. 25]mmol/L) (P ＜ 0. 05 ). Creatinine clearance rates (Ccr) in group D ( [0. 89 ± 0. 19] ml/min ) decreased significantly compared to group C( [1.27 ±0. 33] ml/min) ,as well as group X,Z and L( Ps ＜ 0. 05 ). UAER in group D was significantly higher than that in group C ( [19. 87 ±3. 85] μg/24 h vs [3. 67 ± 1.01] μg/24 h) (P ＜ 0. 05 ), as well as group X, Z and L ( P ＜ 0. 05 ), and the improvement in group L was particularly significant ( P ＜ 0. 05 ). The projections of podocyte in group D severely syncretized, there were slightly improvement in group X, Z and L compared to group D, and the improvement in group L was remarkable. Conclusion The treatment with valsartan, simvastatin and their combination will effectively protect the kidney in early diabetic nephropathy rats,and the effect of using the combination therapy is much better.     

连续性非卧床腹膜透析1个月后患者透析充分性评估及电解质变化

目的 观察连续性非卧床腹膜透析(CAPD)患者治疗1个月的疗效,为早期控制尿毒症患者症状提供较好的方法.方法 新插管后进入CAPD 1个月后的终末期肾脏病患者129例,于CAPD治疗1个月末时评估患者的透析充分性,比较腹透前及第1个月末时观察合并症情况及各生化指标.结果 患者CAPD 1个月末时透析充分性良好,与腹透前比较,CAPD 1个月后水肿的发生率显著下降(24.8%与7.8%,χ2=13.765,P＜0.05),恶心、呕吐等胃肠道不适症状发生率显著下降(66.7%与6.2%,χ2=101.821,P＜0.05),皮肤瘙痒发生率显著下降(22.5%与6.2%,χ2=13.914,P＜0.05),合并症较透析前明显减少;治疗前后对比,血红蛋白[(79.10±17.13)g/L与(96.50±18.69)g/L,t=-6.333,P＜0.01]显著改善,血钙[(1.99±0.30)mmol/L与(2.07±0.20)mmol/L,t=-1.920,P＞0.05]、白蛋白[(30.62±5.24)g/L与(31.84±5.64)g/L,t=-0.333,P＞0.05],血磷[(2.06±0.54)mmol/L与(1.72±0.52)mmol/L,t=3.284,P＜0.01]、血钾[(4.30±0.68)mmol/L与(3.84±0.47)mmol/L,t=4.669,P＜0.01]均较透析前降低,尿素氮[22.00(15.87,30.01)mmol/L与17.00(13.91,20.91)mmol/L,Z=-3.717,P＜0.01]、肌酐[864.00(733.00,1046.25)μmol/L与777.50(627.00,1047.75)μmol/L,Z=-2.408,P＜0.05]均较透析前显著降低.甲状旁腺素[184.80(114.21,369.77)ng/L与226.26(124.22,335.92)ng/L,Z=-0.597,P＞0.05]有所上升,但差异无统计学意义.结论 CAPD在透析早期疗效显著,透析充分性良好,低钙、高磷得到改善,血钾降低,患者生活质量明显改善.

Abstract：

Objective To investigate the impact of continued ambulatory peritoneal dialysis (CAPD)for 1 month,thus to provide effective therapy to control the symptoms of uremia in early stage. Methods A total of 129 nephrotic patients in final stage were treated with CAPD ,dialysis adequacy were assessed after 1 month of CAPD. Complications and biochemical indicators were compared between before and after 1 month of CAPD. Results The dialysis adequacy was good at the end of 1 month of CAPD. Compared to before CAPD,The prevalence of edema after 1 month of CAPD significantly decreased compared to before CAPD (7.8%vs. 24.8% ,χ2 = 13.765, P ＜ 0.05 ). After CAPD gastrointestinal, symptom, such as nausea and vomit significantly decreased from 66.7% to 6. 2% ( χ2 = 101. 821, P ＜ 0. 05 ). Itch of skin significantly decreased from 22. 5% before CAPD to 6. 2% after CAPD(χ2 = 13.914,P ＜0. 05) . Hemoglobin increased significantly from (79. 10 ± 17.13 ) g/L to (96. 50 ± 18. 69 ) g/L after CAPD ( t = - 6. 333, P ＜ 0. 01 ), serum calcium was sisilar, ( 1.99 ± 0.30) mmol/L and (2.07 ± 0. 20) mmol/L at before and after CAPD respectively ( t = -1. 920,P ＞0. 05). Albumin was (30. 62 ±5.24) g/L before CAPD and after CAPD(31.84 ±5.64) g/L ,with no significant difference ( t= - 0.333, P ＞ 0. 05 ) . Serum inorganic phosphorus, kalemia, urea nitrogen and creatinine concentration significantly decreased from ( 2. 06 ± 0. 54 ) mmol/L, ( 4.30 ±: 0. 68 ) mmol/L, 22. 00( 15.87,30.03 ) mmol/L and 864. 00 ( 733.00,1046. 25 ) μmol/L to ( 1.72 ± 0. 52) mmol/L, ( 3.84 ± 0.47 )mmol/L , 17.00 ( 13.91,20. 91 ) mmol/L and 777. 50 ( 627.00, 1047.75 ) μnol/L, respectively ( t = 3.284,4. 669, Z = - 3.717 and - 2. 408, respectively,Ps ＜ 0. 01 or 0. 05 ).. The level of serum PTH increased slightly from [ 184. 80 ( 114. 21,369. 77) ng/L to 226. 26 ( 124. 22,335.92 ) ng/L, but the difference was not significant ( Z = - 0. 597, P ＞ 0. 05 ). Conclusion CAPD had significant effect in early stage of dialysis with good dialysis adequacy. Hypocalcemia and hyperphosphatemia can be improved. The levels of serum kalemia decreased. The iatients's quality of life significantly improved.     

辛伐他汀对非对称性二甲基精氨酸引起的内皮细胞炎症反应的抑制作用

目的:观察辛伐他汀对非对称性二甲基精氨酸(ADMA)引起的人脐静脉内皮细胞(HUVECs)炎症反应的抑制作用,探讨他汀类药物除降脂作用以外的其他抗动脉粥样硬化机制.方法:体外培养的人济静脉内皮细胞,待细胞生长到融合状态时加入不同浓度的ADMA(3、10、30μmol/L)作用48h,观察ADMA对内皮细胞炎症因子产生的影响.不同浓度的辛伐他汀预先孵育20min,然后加入ADMA(30 μmol/L)作用48h,用ELISA法检测上清液中MCP-1、IL-6和TNF-a浓度,比色法检测一氧化氮(NO)含量的变化.结果:ADMA呈剂量依赖性增加上清液中MCP-1、IL-6和TNF-α浓度并降低NO的含量.外源性补充辛伐他汀可逆转ADMA的效应,且呈剂量依赖性(P＜0.05).用一氧化氮合酶抑制剂左旋硝基精氨酸甲酯(L-NAME)抑制一氧化氮合酶降低NO的合成后,可部分取消辛伐他汀的保护作用.结论:ADMA通过诱导炎症反应,引起内皮功能紊乱,其紊乱程度与ADMA的浓度有关;而辛伐他汀能以剂量依赖的方式抑制ADMA诱导的炎症反应,其机制除与其增加NO的合成有关外,可能还有其他物质或途径的参与.     

非对称性二甲基精氨酸和肿瘤坏死因子对原发性高血压左心室构型的影响

目的 观测正常人及高血压患者在不同左心室构型组血清中的肿瘤坏死因子α(TNF-α)和非对称性二甲基精氨酸(ADMA)浓度变化,并探讨TNF-α和ADMA在高血压心脏损害中的病理生理作用.方法 选取体检健康人员30名为正常对照组.轻中度高血压患者66例,根据左心室质量指数(LVMI)和相对室壁厚度(RWT)将其分为4组,即正常左心室构型组:LVMI和RWT均正常;向心性构型组:LVMI正常,RWT增大;向心性肥厚型组:LVMI和RWT均增大;离心性肥厚型组:LVMI增大,RWT正常.采用放射免疫法及反相高效液相色谱法分别测定各组血清中TNF-α及ADMA.结果 对照组和以上4组中TNF-α分别为(11.86±2.45)、(22.08±4.67)、(25.88.4±5.36)、(32.54±5.63)、(48.72±8.86)ng/L;ADMA:(0.78±0.22)、(1.32±0.18)、(1.87±0.20)、(3.03±0.14)、(4.11±0.17)p.mol/L.血清TNF-α和ADMA水平随左心室构型严重程度的增加而升高,左心室构型组患者均明显高于正常对照组(P均<0.01),且左心室重构各亚组间两两比较差异均有统计学意义(P均<0.01).结论 TNF-α和ADMA可能参与了高血压左心室构型的发生和发展.     

非对称性二甲基精氨酸对内皮生长晕细胞凋亡和功能的影响

目的:探讨非对称性二甲基精氨酸(ADMA)对内皮生长晕细胞(EOCs)凋亡和功能的影响.方法:密度梯度离心法分离脐血单个核细胞,培养并扩增EOCs,免疫组化法、荧光染色法鉴定其内皮细胞特性.将浓度为0、1、5、10、30μmol/L的ADMA与EOCs作用48 h,流式细胞仪检测细胞凋亡率,DAPI染色观察凋亡细胞核形态变化,并在倒置显微镜下检测细胞的黏附能力和成血管能力.结果:ADMA(1～30μmol/L)呈浓度依赖性的诱导EOCs的凋亡(P<0.01).ADMA处理组于DAPI染色下可见更显著的细胞凋亡形态学改变.除1μmol/L ADMA外,5、10、30 μmol/L ADMA均可抑制EOCs的黏附能力.10 μmol/L ADMA作用组与对照组相比,明显降低细胞成血管能力(P<0.01).结论:ADMA可以诱导体外培养的EOCs发生凋亡并抑制其黏附能力和成血管能力.     

Clinical study of the changes in homeostasis during the course of acute poisoning in patients treated with hemoperfusion北大核心CSCD

Objective To observe the effects of hemoperfusion on homeostasis in patients with acute poisoning. Methods The data of 26 acute poisoning patients treated with hemoperfusion were retrospective analyzed. The clinical data included blood pH, PvCO2, PvO2, blood lactate, potassium, free-calcium, bicarbonate and blood glucose assayed and recorded at 0 min, 30 min and 120 min after hemoperfusion. The statistical software SPSS 18. 0 was utilized to analyze the statistical differences in the above biomarkers among three different intervals after hemoperfusion. Results At the beginning of hemoperfusion therapy, levels of homeostasis indicators were pH (7. 36 ± 0. 05), PvCO2 (41. 0 ± 8. 8) mmHg, PvO2 (37. 0 ± 11. 8) mmHg, lactate (1.35 ± 1.00) mmol/L, potassium (3.1 ±0.5) mmol/L, sodium (136.3 ± 4.8) mmol/L, free-calcium (0.95 ±0.11) mmol/L, blood glucose (7.90 ±3.47) mmol/L, bicarbonate (22. 8 ± 3. 3) mmol/L. At 30 min, the levels of those were (7. 36 ± 0. 04), (40. 0 ± 5. 7) mmHg, (41.0±7.5) mmHg, (1.11±0.57) mmol/L, (3.1 ±0.4) mmol/L, (137.3±5.4) mmol/L, (0. 94 ±0. 12) mmol/L, (6. 20 ± 1. 55) mmol/L, (22. 2 ±2. 3) mmol/L, respectively. At 120 min, the levels of those were (7. 35 ± 0. 06), (38. 0 ± 6. 7) mmHg, (46. 0 ± 7. 9) mmHg, (0. 69 ± 0. 52) mmol/L, (3.0±0.4) mmol/L, (137.3±5.0) mmol/L, (0.97±0.10) mmol/L, (5.88±1.43) mmoL/L, (22. 0±2. 2) mmol/L, respectively. Apparently, there were significant statistical difference in PvO2 lactate and blood glucose (P < 0. 05) among three different intervals, and no significant statistical differences in other indicators (P > 0.05). Conclusions There were no significant effects of hemoperfusion on relevant indicators in acute poisoning patients.     

内源性一氧化氮合酶抑制物与肺动脉高压研究进展

内源性一氧化氮合酶抑制物非对称二甲基精氨酸(ADMA)是近年来的研究热点,被认为是一种新的心血管疾病风险因子。其代谢主要经二甲基精氨酸二甲胺水解酶(DDAH)水解途径完成。ADMA/DDAH通路的失调可能引起内皮功能不全、肺血管重构等一系列病理改变并导致肺动脉高压形成。阐明ADMA在肺动脉发生发展中的作用对肺动脉高压防治具有重要意义。     

原发性高血压患者血浆ADMA浓度外周循环内皮祖细胞数量的变化

目的探讨原发性高血压（EH）患者血浆非对称性二甲基精氨酸（ADMA）浓度、外周循环内皮祖细胞（EPCs）数量的变化及两者之间的关系。方法选取停止服用降压药至少2周以上并排除冠状动脉粥样硬化性心脏病、糖尿病的原发性高血压患者46例,对照组25例为健康体检者。采用反相高效液相色谱法（RP—HPLC）测定血浆ADMA含量,流式细胞术分析外周循环EPCs。结果EH组血浆ADMA浓度[（0.363±0．029）μg/ml]显著高于对照组[（0．319±0．022）μg／ml]（P〈0．01）。EH组外周循环EPCs数量[（0．368±0．121）％]显著低于对照组[（0．588±0．139）％]（P〈0．01）。血浆ADMA浓度与外周循环EPCs数量呈负相关关系（r=-0．706,P〈0．01）。结论EH患者血浆ADMA浓度升高,外周循环EPCs数量减少,两者之间呈负相关关系。EH患者循环EPCs数量下降部分原因可能是由于血浆ADMA水平升高所致。     

甲状腺激素通过调节甲亢大鼠卵巢葡萄糖转运影响黄体增殖和血管生成、凋亡的机制

目的 探究甲状腺激素通过调节甲亢大鼠卵巢葡萄糖转运影响黄体增殖和血管生成、凋亡的机制.方法 将无特定病原体(SPF)级3周龄Sprague-Dawley大鼠根据随机数字表法分为两组:对照组(大鼠每日灌服5 mL蒸馏水,n=5)和甲亢组(L-甲状腺素250μg/kg溶于5 mL蒸馏水后灌胃,诱导大鼠甲亢,n=5).甲亢诱...     

叔丁基过氧化氢体外诱导鼠系膜细胞衰老及普罗布考对其影响的研究

目的 探讨叔丁基过氧化氢体外诱导鼠肾小球系膜细胞衰老及普罗布考延缓衰老的作用.方法 应用不同浓度叔丁基过氧化氢刺激鼠系膜细胞,每天刺激1 h,连续作用4 d.刺激结束后48 h检测细胞存活率、β-半乳糖苷酶染色阳性细胞率、细胞周期情况鉴定衰老细胞,并透射电镜观察衰老细胞超微结构,并观察普罗布考对上述指标的影响.结果 30μmol/L叔丁基过氧化氢诱导组细胞存活率为对照组的(80.12±3.25)%(P<0.05),约81%的细胞呈现β-半乳糖苷酶染色阳性,流式细胞仪检测86%细胞阻滞于G0、G1期,透射电镜可见叔丁基过氧化氢诱导组细胞核膜内陷,染色质凝聚.应用普罗布考后细胞存活率为对照组的(92.68±5.03)%,β-半乳糖苷酶染色阳性率降至45.2%,细胞周期各期比例接近正常,细胞形态及超微结构改变减轻.结论 叔丁基过氧化氢可诱导体外培养的鼠肾小球系膜细胞发生衰老,应用普罗布考可以延缓衰老.     

诺和龙治疗2型糖尿病合并动脉粥样硬化患者的疗效观察

目的 观察诺和龙治疗2型糖尿病合并动脉粥样硬化患者的临床疗效.方法 将65例2型糖尿病合并动脉粥样硬化患者随机分为诺和龙组(36例)和优哒灵组(29例),诺和龙组患者降糖药均为诺和龙治疗,优哒灵组均为优哒灵治疗,观察疗程12个月,监测治疗前后的血糖含量和颈动脉内膜中层厚度.结果 诺和龙组能有效降低餐后血糖(1.99±1.06)mmol/L,优哒灵降低餐后血糖(0.99±0.54)mmol/L,2组比较差异有统计学意义(P＜0.05).诺和龙组治疗后颈动脉内膜中层厚度比治疗前减少[(1.02±0.08)、(1.11±0.07)mm](P＜0.05),优哒灵组治疗后颈动脉内膜中层厚度与治疗前比较无差异[(1.07±0.06)、(1.10±0.08)mm](P＞0.05).结论 诺和龙是一种安全、有效的降糖药物,适于2型糖尿病合并动脉粥样硬化患者的治疗.     

急性脑梗死患者溶栓治疗前后血浆基质金属蛋白酶-9含量的变化及意义

目的 探讨血浆基质金属蛋白酶-9(MMP-9)在急性脑梗死患者溶栓治疗后的变化及临床意义.方法 测定34例急性脑梗死患者溶栓前后血浆MMP-9水平,并与健康对照组(34例)比较.结果 急性脑梗死患者溶栓前MMP-9水平较健康对照组无明显升高[(13.47±3.09)ng/L比(12.89±10.22)ng/L,P＞0.05],溶栓后MMP-9水平[(22.06±12.53)ng/L]较健康对照组和溶栓前均显著升高(均P＜0.05).溶栓后发生出血患者(发生率26.5%,9/34)MMP-9水平较溶栓前显著增加[(24.02±15.41)ng/L比(14.28±2.33)ng/L,P＜0.05];与无出血患者[(20.42±9.57)ng/L]相比有增高趋势,但差异无统计学意义(P＞0.05).溶栓后完全再通患者(再通率58.8%,20/34)MMP-9水平较溶栓前明显升高[(19.26±7.94)ng/L比(13.63±3.02)ng/L,P＜0.05];与不完全再通患者[(18.97±4.23)ng/L]相比有增高趋势,但差异无统计学意义(P＞0.05).结论 溶栓后激活了MMP-9,MMP-9增加溶栓后出血的风险并参与了溶栓后出血的机制.     

血液透析患者肺动脉高压与血管内皮功能紊乱的相关性

目的 研究维持性血液透析（maintenance hemodialysis,MHD）患者肺动脉高压（pulmonary hypertension,PAH）与内皮细胞功能紊乱之间的关系.方法 选取维持性血液透析患者60名,行心脏彩超检查,以肺动脉收缩压（PASP）≥35mmHg的为肺动脉高压组（PAH组）,PASP＜35 mmHg的为无肺动脉高压组（无PAH组）,分别收集2组患者临床资料和实验室数据,在透析间期测量2组患者血流介导的肱动脉内皮依赖性舒张功能（FMD）及人血不对称二甲基精氨酸（ADMA）浓度.结果 60例患者17人存在PAH（28.3％）,PAH组和无PAH组患者透析间期体质量增加量与干体质量比值及高敏C反应蛋白（hs-CRP）存在显著差异（P＜0.05）.经检测PAH组和无PAH组FMD分别为（12.2±1.3）％和（6.9±1.2）％,ADMA分别为（2.97±0.31）μmol/L和（2.i0±0.29）μmol/L,采用协方差分析法矫正可能和血管内皮功能相关的其他因素后,得出FMD和ADMA在两组之间均有显著差异（F=63.8,P＜0.001,F=16.832,P＜0.01）.二分类Logistic回归分析显示,左心室质量分数（LVMI） （B=0.037,P=0.043）和ADMA （B=9.519,P=0.006）是MHD患者并发PAH的主要相关因素.结论 MHD合并PAH的患者存在显著的血管内皮细胞功能紊乱,LVMI和人血ADMA是MHD患者并发PAH的主要相关因素.     

阿托伐他汀对急性冠状动脉综合征患者不对称二甲基精氨酸的影响

目的 研究阿托伐他汀对急性冠状动脉综合征(ACS)患者不对称二甲基精氨酸(ADMA)的影响及意义.方法 随机选择ACS患者78例,入选后再随机分为阿托伐他汀组(他汀组)39例,常规治疗组(常规组)39例;随机选取我院同期的健康体检者40例作对照组.测定每组治疗前后血脂和ADMA水平.结果 ①急性心肌梗死(AMI)组ADMA浓度(1.09±0.36)μmol/L高于不稳定型心绞痛(UAP)组(0.91±0.22)μmol/L(P<0.01),AMI组和UAP组ADMA浓度高于正常对照组(0.57±0.12)μmol/L(P<0.01).②他汀组经阿托伐他汀治疗后,ADMA、总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)均低于治疗前,分别为ADMA(0.67±0.25)μmol/L vs(1.02±0.32)μmol/L,TC(5.26±0.78)mmol/L vs(6.86±1.09)mmol/L,TG(1.58±0.45)mmol/L vs(2.06±0.37)mmol/L,LDL-C(2.51±0.48)mmol/L vs(2.97±0.45)mmol/L(P<0.01);常规组治疗后以上指标降低不多,他汀组治疗后ADMA、TC、TG、LDL-C均低于常规组.结论 阿托伐他汀可能通过调节ACS患者血浆血脂水平,进而降低ADMA的生成.     

C5aR和P38-MAPK在脓毒性休克大鼠心肌损伤中的作用

目的 研究补体C5a受体与P38-MAPK在脓毒性休克情况下诱导心肌损伤的关系.方法 采用盲肠结扎剪切法构建早期脓毒性休克大鼠动物模型,实验地点在武汉大学人民医院麻醉科实验室.30只Sprague-Dawley大鼠随机(随机数字法)分为正常对照组6只和模型组24只(12 h组12只,24 h组12只).12 h组于术后12 h时点检测血清乳酸脱氢酶(LDH)和肌酸激酶(CK)水平,12 h后麻醉处死大鼠,迅速开胸取出心脏组织,行HE染色、应用免疫组织化学方法检测C5a受体和P38-MAPK的表达情况.24 h组亦于术后24 h时点检测血清LDH和CK值,24 h后处死大鼠取心肌组织行HE染色、检测C5a受体和P38-MAPK的表达情况.结果 模型组(12 h组和24 h组)与正常对照组相比,LDH值和CK值均明显升高(P＜0.05).24 h组LDH值和CK值与12 h时间点相比,差异具有统计学意义[(2 568.9±280)vs.(2 201.2±149),(5 029.7±458)vs.(2 629.4±140),P＜0.05].C5aR和P38-MAPK灰度值分析显示,模型组与正常对照组相比,均有显著性增高(P＜0.05),24 h组较12 h组相比,差异具有统计学意义[(702.77±122)vs.(388.36±113),(646.40±181)vs.(307.32±61),P＜0.05].相关分析显示C5aR和P38-MAPK存在显著正相关关系,(P＜0.05),P38-MAPK与LDH和CK均存在显著正相关关系(P＜0.05).结论 C5a受体和P38-MAPK在诱导脓毒性休克心肌损伤中有着强大的协同效应.

Abstract：

Objective To investigate effects of complement C5a receptor and P38-MAPK on myocardial injury brought about by septic shock in rats. Method The early septic shock models were established by the method of cecal ligature and incision (CLI). A total of 30 Sprague-Dawley rats were randomly( random number) divided into normal control group ( n = 6 ) and model group ( n = 24 ) and the model group was further 12 hours later divided into 12 h subgroup (n = 12) and 24 h subgroup (n = 12). The arterial blood samples were collected 12 hours later for detecting the levels of lactate dehydrogenase (LDH) and creatine kinase (CK), and then the rats were sacrificed and the myocardial tissues were taken to assay the expressions of C5a receptor and P38-MAPK by using immunohistochemistry after HE staining. And the above procedure as did in 12 h subgroup was done 24 hours later. Results Compared with the control group, the levels of LDH and CK in rats of Model group were significantly higher (P ＜ 0. 05). There were significant differences in LDH and CK between 24 h subgroup and 12 h subgroup [(2 568.9 ± 280) vs. (2 201.2 ± 149)] and [(5 029.7±458) vs. (2 629.4±140)] ,P＜0. 05, P＜0.05. The analysis of C5aR and P38-MAPK gray values showed that there were significant differences between the model group and normal control group [(702.77 ±122) vs. (388.36±113)], P＜0. 05 and [(646.40±181) vs. (307.32 ±61)] ,P＜0.05,and those differences also found between the 24 h subgroup and 12 h subgroup. There was a significant positive correlation between C5aR and P38-MAPK (P＜0.05 ), and also the P38-MAPK had significant positive relationships with LDH(P＜0.05) and CK (P＜0.05). Conclusions The C5aR strongly potentiates the P38-MAPK to induce myocardial injury by septic shock.