中药复方调控GRP78、CHOP改善代谢综合征大鼠内质网应激的实验研究

目的 探讨中药复方保护代谢综合征(MS)大鼠肾功能作用的分子机制。方法 将80只Wistar大鼠随机分为对照组、模型组、中药组和小檗碱组,每组20只。模型组、中药组和小檗碱组以10%果糖造模,中药组以中药复方灌胃,小檗碱组以小檗碱灌胃,对照组及模型组给予等体积生理盐水灌胃。8周后测量各组大鼠生化指标、胰岛功能、收缩压;之后处死大鼠,取肾组织行肾脏病理染色及实时定量聚合酶链式反应(RT-PCR)检测各组大鼠葡萄糖调节蛋白78(GRP78)、内质网应激蛋白(CHOP) mRNA表达。结果 模型组大鼠空腹胰岛素水平(FINS)、三酰甘油(TG)、总胆固醇(TC)、血尿酸(UA)均高于对照组,肾组织GRP78、CHOP mRNA表达较对照组增加;中药组、小檗碱组大鼠FINS、TG、TC、UA较模型组均下降,肾组织GRP78、CHOP mRNA表达下降,其中,中药组各指标下降更显著。结论 中药复方通过减轻MS诱导的内质网应激,从而发挥对肾脏细胞损伤的保护作用。     

磷酸肌酸钠对慢病毒介导Calumenin蛋白沉默阿霉素损伤心肌细胞内质网应激信号通路的作用

目的:探讨磷酸肌酸钠对网腔钙结合蛋白(Calumenin)沉默阿霉素损伤心肌细胞内质网应激信号通路作用。方法:培养乳鼠心肌细胞,构建稳定的慢病毒——Calumenin质粒,转染乳鼠培养心肌细胞,实验分为4组:对照组(正常细胞+3mg/L阿霉素)、模型组(慢病毒感染细胞+3mg/L阿霉素)、磷酸肌酸钠1组(正常细胞+阿霉素+磷酸肌酸钠)、磷酸肌酸钠2组(转染细胞+阿霉素+磷酸肌酸钠)。采用Western blotting技术检测各组心肌细胞Calumenin蛋白、内质网应激伴侣蛋白GRP78及内质网应激信号通路因子PERK、PATF-4PERK、eIF2ɑ、ATF-4、IRE1、CHOP表达。结果:1与对照组比较,模型组及磷酸肌酸钠2组心肌细胞Calumenin蛋白表达明显减少(P0.01)。2与对照组相比,模型组内质网应激伴侣蛋白GRP78及内质网应激信号通路因子PPERK、eIF2ɑ、ATF-4、IRE1、CHOP表达明显增多(P0.01)。3与模型组相比较,磷酸肌酸钠1组及磷酸肌酸钠2组内质网应激伴侣蛋白GRP78及内质网应激信号通路因子P-PERK、eIF2ɑ、ATF-4、IRE1、CHOP表达减少(P0.01)。结论:阿霉素损伤可能诱发ERS,并通过ERS凋亡信号通路PERK→P-PERK→eIF2a→ATF-4→CHOP/IRE1→CHOP引起心肌细胞凋亡;磷酸肌酸钠可抑制阿霉素损伤所诱导内质网应激介导的心肌细胞凋亡,这一作用机制可能是通过Calumenin蛋白抑制ERS及其凋亡信号通路PERK→P-PERK→eIF2a→ATF-4→CHOP/IRE1→CHOP实现的。     

脂联素抑制内质网应激干预慢性间歇性缺氧所致肾损伤

目的探讨脂联素(Ad)对慢性间歇性缺氧(CIH)所致肾损伤的干预作用及相关机制。方法 60只成年Wistar大鼠随机分成4组:正常对照(NC)组、NC+Ad组、CIH组和CIH+Ad组。其中CIH组和CIH+Ad组大鼠接受CIH处理4个月。其余2组接受正常空气处理,同时NC+Ad组和CIH+Ad组大鼠接受Ad(10l人開)治疗,每周2次,持续4个月。荧光显微镜下观察活性氧(ROS)的水平。TUNEL染色检测肾脏细胞凋亡情况。Western blotting检测各组大鼠肾脏组织中GRP78、CHOP、IRE1、PERK、pro-ATF6蛋白的表达,以反映各组大鼠内质网应激情况。采用SPSS 17.0软件进行统计学分析。计量资料用均数±标准差(±s)表示,组间比较用t检验。结果实验满4个月时,组间比较示各参数在NC与NC+Ad组间差异均无统计学意义(P均0.05)。与NC组和NC+Ad组比较:(1)ROS水平在CIH组显著增高,而CIH+Ad组低于CIH组,但仍高于NC组和NC+Ad组(尸均0.05);(2)肾细胞凋亡率和反映凋亡的caspase-12和caspase-3蛋白水平在CIH组明显增加,CIH+Ad组较CIH组明显减少,但仍然高于NC组和NC+Ad组(P均0.05);(3)CIH组肾脏组织的GRP78、CHOP、IRE1、PERK蛋白水平明显增加,在CIH+Ad组明显减少,但仍高于NC组和NC+Ad组(P均0.05);pro-ATF6蛋白水平在CIH组明显降低,在CIH+Ad组有所增加,但仍然低于NC组和NC+Ad组(P均0.05)。结论 CIH可以通过激活ROS和ERS相关的细胞凋亡途径导致肾脏损伤,而补充外源性Ad后,可能通过抑制ROS,进而抑制ERS,保护肾脏细胞。     

替米沙坦对糖尿病大鼠肾脏组织中内质网应激介导相关细胞凋亡的保护作用

探讨血管紧张素Ⅱ受体拮抗剂对糖尿病大鼠肾脏组织中内质网应激相关的细胞凋亡的影响.31只雄性SD大鼠分为正常对照组、糖尿病组、替米沙坦干预组.12周试验结束后测量大鼠体重、24h尿蛋白量,检测血糖、血胰岛素、血肌酐等.肾脏细胞凋亡用TUNEL法检测;肾脏细胞内质网应激信号通路分子糖调节蛋白78( GRP78)、caspase12和CHOP用免疫组化及实时定量PCR法检测.糖尿病组的血糖、24h尿蛋白量、血肌酐显著高于对照组(P＜0.05);体重和血胰岛素较对照组低(P＜0.05).替米沙坦干预组的24h尿蛋白量、血肌酐较糖尿病组明显减少(P＜0.05),凋亡指数也显著低于糖尿病组(P＜0.05).大鼠内质网应激信号通路分子GRP78、caspase12、CHOP蛋白及其mRNA的表达,糖尿病组显著高于对照组和替米沙坦干预组(P＜0.05).内质网应激参与了糖尿病大鼠肾脏细胞的凋亡,替米沙坦对内质网应激介导相关的肾脏细胞凋亡有保护作用.     

芹黄素对布雷菲德菌素诱导的神经元样PC12细胞内质网应激途径的影响

目的探讨芹黄素对布雷菲德菌素(BF)A诱导的PC12分化细胞内质网应激(ERS)标志分子葡萄糖调节蛋白(GRP)48及ERS相关促凋亡因子C/EBP环磷酸腺苷反应元件结合转录因子同源蛋白(CHOP)表达水平的影响。方法以神经生长因子(NGF)诱导PC12分化细胞作为模型,流式细胞仪检测不同浓度BFA诱导的PC12分化细胞凋亡情况,MTT法检测芹黄素及BFA作用后细胞存活率,PT-PCR及Western印迹检测芹黄素及BFA作用后GRP78及CHOP mRNA及蛋白表达水平。结果 BFA(0.05、0.10、0.50、1.00、5.00、10.00μmol/L)可诱导PC12分化细胞凋亡,激活ERS凋亡途径。芹黄素(10.00、20.00、40.00μmol/L)预处理可使BFA(1μg/ml)诱导的细胞损伤减轻,使细胞存活率明显提高(P<0.05)。芹黄素(10.00、20.00μmol/L)可抑制BFA(1μg/ml)诱导的GRP78及CHOP表达上调(P<0.05)。结论芹黄素对BFA诱导的ERS损伤有保护作用,抑制GRP78/CHOP信号通路可能是其机制之一。     

低血糖对急性心肌梗死后内质网应激介导心肌细胞凋亡的作用和机制研究

目的通过体内大鼠模型观察低血糖水平对内质网应激(ERS)感受蛋白及凋亡信号分子表达的影响。方法将Wister大鼠40只中随机选20只结扎左冠状动脉前降支建立大鼠急性心肌梗死(AMI)模型;另20只为假手术,再随机分为正常血糖假手术组(SN组)、正常血糖AMI组(MN组)、低血糖假手术组(SL组)、低血糖AMI组(ML组),每组10只。建立术前低血糖模型,术后24h处死取材,评估大鼠心肌细胞凋亡程度、检测心肌组织中葡萄糖调节蛋白78/免疫球蛋白结合蛋白(GRP78/BIP)、蛋白激酶R样内质网激酶(PERK)和C/EBP同源蛋白(CHOP)及Caspase-12的表达情况。结果 ML组及MN组心肌细胞凋亡指数[(29.36±2.15)%、(20.27±2.80)%]明显高于SN组和SL组[(1.82±0.83)%、(1.97±0.96)%,P<0.05];MN组和ML组GRP78、PERK、CHOP及Caspase-12的表达明显高于SN组和SL组(P<0.01);MN组与ML组PERK表达比较,无统计学差异(P>0.05)。结论 AMI前低血糖水平激活了ERS诱导的细胞凋亡通路;内质网特异性标记蛋白表达与心肌细胞凋亡变化规律一致,ERS通路可能参与了大鼠AMI后心肌细胞凋亡的调控。     

米诺环素通过调节葡萄糖调节蛋白78的表达抑制糖尿病肾细胞凋亡

目的探讨米诺环素对糖尿病肾损害的保护及与内质网应激的关系。方法 STZ腹腔注射制备糖尿病大鼠模型,分别在成模即刻和8 w后每日给予米诺环素灌胃(20 mg/kg),16 w病程结束检测肾功能。Tunel检测肾组织细胞凋亡指数,免疫组化方法观察病理学改变和GRP78蛋白在肾组织中的表达变化。结果病程持续16 w结束时,DM模型组大鼠与正常对照组相比,内生肌酐清除率(Ccr)降低,血尿素氮(BUN)、24 h尿蛋白(Upro)显著升高;米诺环素干预组Ccr增加,BUN和Upro下降,与模型组相比差异显著。正常对照组的肾组织GRP78呈弱表达,DM模型组大鼠的肾组织中GRP78蛋白表达为强阳性,DM模型组GRP78表达对正常对照组相比有显著性差异。米诺环素干预组,GRP78表达明显下降,而且半程干预组比全程干预组GRP表达下降更明显。正常对照组凋亡细胞偶见;DM模型组肾组织可见大量凋亡细胞,尤其是肾小管上皮,肾小球可见部分凋亡细胞。米诺环素干预组肾小管上皮凋亡细胞明显减少,半程干预组最低。结论米诺环素改善糖尿病肾功能可能部分通过降低肾组织过度激活的内质网应激反应,减少肾小管细胞的细胞凋亡率而实现的。     

贝那普利对糖尿病肾病大鼠肾组织MCP-1表达的影响

目的 观察单核细胞趋化蛋白-1(MCP-1)在早期糖尿病肾病(DN)大鼠肾组织的表达、尿中的排泄情况及贝那普利的干预效果.方法成年雄性Wistar大鼠分为对照组(NC)、DN模型组(DN)和DN 治疗组(DN 贝那普利).DN 贝那普利组给予贝那普利灌胃,NC和DN组以等量蒸馏水灌胃.饲养4 w后观察大鼠生理生化指标的变化、尿中的MCP-1在排泄量、肾脏组织病理学变化及肾脏组织中MCP-1的表达水平.将第5代培养的肾小球系膜细胞分为对照组(NC)、高糖组(HG)和高糖 治疗组(HG 贝那普利),观察系膜细胞内MCP-1 mRNA表达.结果 DN组和DN 贝那普利组大鼠血糖较NC组显著升高(P＜0.01). DN组大鼠肾重、肾脏肥大指数、尿白蛋白(UAlb)/尿肌酐(Ucr)、尿MCP-1/Ucr及肾组织内MCP-1的水平均明显高于NC组;与DN组比较,上述指标在DN 贝那普利组明显下降,差异有统计学意义.肾脏病理显示DN组病理改变较DN 贝那普利组重.细胞实验显示,HG组所培养的系膜细胞内MCP-1 mRNA表达水平显著高于NC组,但用贝那普利处理后MCP-1 mRNA表达水平明显降低(P＜0.01).结论 MCP-1在早期DN大鼠肾组织中的表达及尿中的排泄均增加,MCP-1的监测有望成为早期DN的预测指标.     

运动训练对大鼠脑缺血再灌注后神经细胞内质网应激的影响

目的探讨运动训练对大鼠脑缺血再灌注后神经细胞内质网应激的影响。方法清洁级SD大鼠100只,采用随机数字表法将SD大鼠分为正常对照组、假手术组、模型组和实验组,每组25只。造模成功后,实验组进行功能训练。于造模后1、3、7、14、28 d,分别处死5只大鼠,取脑组织,采用TUNEL检测神经细胞凋亡情况,采用RT-CPR和Western印迹法检测葡萄糖调节蛋白(GRP)78、C/EBP同源蛋白(CHOP)和Tribbles相关蛋白(TRB)3表达水平。结果缺血再灌注后1、3、7、14、28 d,实验组凋亡细胞数明显少于模型组(P<0.05)。缺血再灌注后1、3、7、14、28 d,实验组和模型组GRP78、CHOP和TRB3 mRNA和蛋白表达水平均显著高于假手术组(P<0.05);缺血再灌注后3 d、7 d、14 d、28 d,实验组大鼠脑组织GRP78、CHOP和TRB3 mRNA和蛋白表达水平显著低于模型组(P<0.05)。结论运动训练能明显抑制脑缺血再灌注后神经细胞凋亡,机制可能主要通过下调脑组织GRP78、CHOP和TRB3表达水平实现的。     

噪声应激对大鼠心肌葡萄糖调节蛋白78和CCAAT/增强子结合蛋白同源蛋白表达的影响与高血压及心肌重构的关系

目的 研究内质网应激相关因子葡萄糖调节蛋白78(GRP78)和CCAAT/增强子结合蛋白同源蛋白(CHOP)在噪声应激环境高血压大鼠模型心肌中的表达,并探讨GRP78和CHOP变化与高血压及其心肌重构的关系.方法 将成年雄性Sprague-Dawly(SD)大鼠40只随机分为两组:噪声组(采用噪声环境建立高血压模型)和...     

Levels of circulating GRP78 and CHOP in endoplasmic reticulum stress pathways in Chinese type 2 diabetic kidney disease patients

The current study aimed to investigate circulating glucose-regulated protein 78 (GRP78) as well as CCAAT/enhancer-binding protein homologous protein (CHOP) concentrations in Chinese type 2 diabetes mellitus (T2DM) patients, especially those with microalbuminuria. We recruited 67 patients with T2DM and 63 control subjects. We determined circulating GRP78 and CHOP concentrations by ELISA, collected anthropometric data, and measured biochemical parameters in a clinical laboratory. Compared with control groups, patients with T2DM showed decreased circulating levels of GRP78 (0.21 [0.16–0.24] vs 0.16 [0.16–0.19] ng/mL, P < .01) and CHOP ([0.29 ± 0.02] vs [0.27 ± 0.03]ng/mL, P < .01). Reduction in circulating GRP78 and CHOP levels was more pronounced in patients with more severe categories of albuminuria. Amounts of circulating GRP78 correlated directly with serum fasting c-peptide, cystatin-c (Cys-c), creatinine (Cr), blood urea nitrogen (BUN), and uric acid, and inversely with glomerular filtration rates. Circulating CHOP level was positively correlated with age, Cr, BUN, Cys-c, and urinary microalbumin/creatinine (UmALB/Cr). Circulating GRP78 was predicted independently by Cr, BUN, serum uric acid, estimated glomerular filtration rate, and Cys-c, while CHOP depended on age, Cr, BUN, estimated glomerular filtration rate, UmALB/Cr, and Cys-c. After controlling for confounding factors, circulating GRP78 and CHOP expression were significantly associated with diabetic kidney disease (binary logistic regression, P < .01). Patients with T2DM showed increased circulating GRP78 and CHOP concentrations. Receiver operating characteristic areas under the curve for predicting diabetic kidney disease based on GRP78 and CHOP were 0.686 (95% CI: 0.558–0.813) and 0.670 (0.524–0.816), respectively.     

Usefulness of procalcitonin for diagnosis of infective endocarditis

The aim of this study was to evaluate the accuracy of procalcitonin (PCT) in predicting infective endocarditis (IE). 23 adult patients with IE, 30 patients with sepsis and 30 with tick-borne encephalitis were included in this prospective study. The PCT serum level, C-reactive protein (CRP), total leukocyte, and immature polymorphonuclear (PMN) cell counts were determined on admission, prior to the institution of antibiotic therapy, and compared according to the diagnosis. The median PCT level in patients with IE endocarditis was 0.81 ng/ml, in patients with sepsis it was 43.74 ng/ml, and in the group with viral infection it was 0.25 ng/ml (P < 0.001). The highest PCT level was found in patients with Staphylococcus aureus endocarditis. The area under the receiver operating characteristic curve that used PCT to predict IE was 0.722 (95% CI 0.572–0.873), compared with 0.909 (95% CI 0.829–0.989) for CRP, 0.699 (95% CI 0.551–0.846) for immature PMN cell count, and 0.619 (95% CI 0.468–0.770) for leukocyte count. Our study fails to demonstrate superiority of PCT as a diagnostic laboratorial parameter in predicting IE compared to CRP.     

大鼠肝癌发生过程中癌基因和抑癌基因的表达

目的 探讨癌基因ｒａｓ和抑制基因Ｐ５３蛋白在实验性肝癌中的表达与内在联系。方法 应用原位杂交和免疫组织化学（ＳＡＢＣ０法,在３８例化学致癌剂二乙基亚硝胺（ＤＥＮＡ）诱发大鼠原发性肝细胞癌及癌前增殖结节中。Ｐ５３蛋白和ｒａｓ家族基因蛋白表达。结果 化学诱癌生成为６８．４２％,癌前增生性病例为３１．５８％,在癌与增生性病变中ｒａｓ基因收搞表达分别为８８．４６％和７５％,两组无显著差异,但与癌的恶笥分化     

Effects of low-dose hydroxychloroquine on expression of phosphorylated Akt and p53 proteins and cardiomyocyte apoptosis in peri-infarct myocardium in rats

BACKGROUND:

Low-dose hydroxychloroquine (HCQ) and ataxia-telangiectasia-mutated (ATM) protein kinase have recently been postulated to be beneficial for the prevention of the age-associated metabolic syndrome including hypertension, hypercholesterolemia and glucose intolerance; however, the effects of low-dose HCQ on the expression of ATM downstream phosphorylated Akt (protein kinase B) and p53 proteins and cardiomyocyte apoptosis in the peri-infarct myocardium remain unclear.

OBJECTIVE:

To explore the effects of low-dose HCQ on the expression of phosphorylated Akt and p53 proteins and cardiomyocyte apoptosis in the peri-infarct myocardium in a rat model.

METHODS:

Myocardial infarction (MI) was induced experimentally in a subset of rats, while others underwent sham operation (sham). Three days after operation, surviving Sprague-Dawley male rats were divided into MI+HCQ, MI, sham+HCQ and sham groups. MI+HCQ and sham + HCQ groups were treated with HCQ (3.4 mg/kg); and MI and sham groups were treated with phosphate buffered (ie, physiological) saline (10 mL/kg) by gavage every day for 12 weeks. The expression of phosphorylated Akt and p53 proteins and cardiomyocyte apoptosis in the peri-infarct myocardium was detected by Western blot and terminal deoxynucleotidyl transferase dUTP nick end labelling, respectively.

RESULTS:

Twelve weeks after treatment, the expression of phosphorylated Akt protein was significantly increased (P<0.05). Expression of phosphorylated p53 protein was not significantly different (P>0.05) in the peri-infarct myocardium of the MI+HCQ group from that in the MI group. The cardiomyocyte apoptosis rate in the peri-infarct myocardium was significantly decreased in the MI+HCQ group compared with the MI group (P<0.05).

CONCLUSION:

Low-dose HCQ can significantly increase the expression of phosphorylated Akt protein without significantly impacting expression of phosphorylated p53 protein in the peri-infarct myocardium. Accordingly, it can inhibit cardiomyocyte apoptosis in the peri-infarct myocardium.     

A diagnostic test: combined detection of heparin-binding protein,procalcitonin, and C-reactive protein to improve the diagnostic accuracy of bacterial respiratory tract infections

BackgroundRespiratory tract infection (RTI) is one of the most common diseases worldwide, and its incidence is rising year by year due to environmental pollution. Sputum culture remains the gold standard for RTI diagnosis, but its performance is limited by difficulties related to the sampling and testing of the sputum specimens. Heparin-binding protein (HBP), procalcitonin (PCT), and C-reaction protein (CRP) are Inflammatory markers. They have the advantage of being fast, accurate and reproducible, but limited by their sensitivity and specificity. We explored the clinical value of the combined detection of them in the diagnosis of bacterial RTIs.MethodsPatients who fulfilled the inclusion criteria were selected as the case group, healthy age- and sex-matched subjects were enrolled as a control group. The subjects’ HBP, PCT, and CRP levels were detected. The case group was further divided into two groups according to the bacterial culture results, and the differences in the markers were statistically analyzed. The receiver operating characteristic (ROC) curves were drawn, and the areas under the ROC curve (AUCs) were calculated to analyze the diagnostic values of each marker and their combination in parallel for bacterial RTIs.ResultsThe plasma HBP, PCT, and CRP levels of patients in the bacterial and non-bacterial infection groups were significantly higher than those of patients in the healthy control group, and were positively correlated to the severity of the disease. for HBP with an AUC of 0.785 [95% confidence interval (CI): 0.686–0.884], a sensitivity of 0.821, a specificity of 0.771; PCT with an AUC of 0.767 (95% CI: 0.664–0.870), a sensitivity of 0.773, a specificity of 0.791, and CRP with an AUC of 0.748 (95% CI: 0.642–0.854), a sensitivity of 0.839, a specificity of 0.696 in the bacterial and non-bacterial infection groups. The combined detection of HBP + CRP had the optimal diagnostic performance, with an AUC of 0.797 (95% CI: 0.698–0.895; P<0.001), a sensitivity of 0.809, a specificity of 0.800.ConclusionsThe combined detection of HBP and CRP is valuable for diagnosing bacterial RTIs and may guide the development of reasonable treatment protocols in clinical settings.     

Significance of measurement of high-sensitivity C-reactive protein in acute pancreatitis

Background: Determination of the severity of acute pancreatitis is difficult in the early phase after onset, and we often encounter difficulties in making decisions to initiate intensive care during the early phase. Therefore, there is real need for a simple and inexpensive method that can precisely evaluate the severity of acute pancreatitis. Methods: In the present study, we measured serum C-reactive protein (CRP) levels in 20 patients with acute pancreatitis, using a high-sensitivity CRP (hs-CRP) assay method. Results: CRP levels were as low as 1.0, 0.4, and 0.3 mg/dl in cases 2, 3, and 9, respectively, with severe acute pancreatitis. These three patients were hospitalized within 24 h after the onset of pancreatitis. Cases 2, 3, and 9 showed high hs-CRP levels, of 209 000, 68 600, and 154 000 ng/ml, respectively, and their interleukin (IL)-6 levels were above 500 pg/ml. The mean hs-CRP level was 222 760 ± 32 197 ng/ml in patients with severe acute pancreatitis and 22 798 ± 8216 ng/ml in patients with mild to moderate pancreatitis, with a significantly higher level in the severe cases. Cases 14, 16, and 20, with mild to moderate pancreatitis, had hs-CRP levels of 83 400, 1800, and 55 400 ng/ml, respectively. Conclusions: Measurement of hs-CRP levels is a simple and inexpensive method. The hs-CRP levels were found to significantly increase in the early phase of severe acute pancreatitis, suggesting that hs-CRP could possibly serve as an early indicator of the progression of acute pancreatitis into a serious state. Received: February 1, 2002 / Accepted: May 31, 2002 Reprint requests to: S. Tanaka     

冠状动脉支架植入术后血浆C反应蛋白水平的评价

目的：测定冠状动脉支架植入（CSI）术后血清C反应蛋白（CRP）水平，了解其意义。方法：选择2000年1月－2001年7月在我院行CSI术患者为研究对象（n=58)，分为A组（阵旧性心肌梗死及心绞痛患者，n=43例），及B组（急性心肌梗死患者,n=15)。A组于CSI术前、后，B组于CSI术当日，术后1，3，8日清晨抽取空腹血进行血清CRP及肌酸激酶同功酶（CK－MB）测定。结果：A组：CSI术后平均血清CRP水平高于术前（P＜0．01），CK－MB较术前无显著性差异；B组：CSI术后CRP及CK－MB水平随时间的推移呈递减趋势。结论：CSI术后血清CRP水平升高；它与局部机械损伤引起的炎症有关，可作为CSI术损伤程度及AMI病程转归的监测指标。     

Diversity in Glycosaminoglycan Binding Amongst hMPV G Protein Lineages

We have previously shown that hMPV G protein (B2 lineage) interacts with cellular glycosaminoglycans (GAGs). In this study we examined subtypes A1, A2 and B1 for this interaction. GAG-dependent infectivity of available hMPV strains was demonstrated using GAG-deficient cells and heparin competition. We expressed the G protein ectodomains from all strains and analysed these by heparin affinity chromatography. In contrast to the B2 lineage, neither the A2 or B1 G proteins bound to heparin. Sequence analysis of these strains indicated that although there was some homology with the B2 heparin-binding domains, there were less positively charged residues, providing a likely explanation for the lack of binding. Although sequence analysis did not demonstrate well defined positively charged domains in G protein of the A1 strain, this protein was able to bind heparin, albeit with a lower affinity than G protein of the B2 strain. These results indicate diversity in GAG interactions between G proteins of different lineages and suggest that the GAG-dependency of all strains may be mediated by interaction with an alternative surface protein, most probably the conserved fusion (F) protein. Analysis of both native and recombinant F protein confirmed that F protein binds heparin, supporting this conclusion.     

Galectin-3在CCl4致肝损伤及对GRP78调控的作用

目的:探讨Galectin-3对CCl_4致急性肝损伤时GRP78的调控作用.方法:选择ICR系的♂Galectin-3基因敲除型[Gal-3(-/-)]和其野生型[Gal( / )1小鼠,一次灌胃给予CCl_4,观察给药后10,24,48和72 h的肝组织病理改变和检测其血清谷草转氨酶(AST)和谷丙转氨酶(ALT)活性,并检测肝细胞不同细胞组分中的GRP78蛋白表达.结果:与Gal( / )型鼠比较,CCl_4对Gal-3(-/-)型鼠的肝组织病理损伤出现时间早、损伤程度重.Gal-3(-/-)型鼠在CCl_4灌胃后10和24 h的血清ALT活性(1 860±191 U/L vs 1356±177 U/L,t=6.12,P<0.01;2789±236 U/L vs 2468±221 U/L,f=3.14,P<0.01)及CCl_4灌胃后10 h的血清AST活性(946±89 vs 623±73 U/L,t=8.87,P<0.01)与Gal( / )型鼠比较显著升高.对照组中Gal-3( / )型鼠的微粒体组分(Mic)中的GRP78蛋白表达显著高于Gal-3(-/-)型鼠(140.9±21.1 vs 76.1±9.5,t=8.86,P<0.01).在CCl_4 ig后24 h,Gal-3( / )型鼠的肝细胞线粒体组分(Mt)和Mic中的GRP78蛋白表达明显升高,并显著高于Gal-3(-/-)型鼠(Mt:127.0±18.8 vs 49.1±6.3,P<0.01;Mic:166.5±23.4 vs 87.7±11.6,P<0.01).结论:Galectin-3蛋白在CCl_4致急性肝损伤中可能具有一定的保护作用.上调Mic和Mt中的GRP78蛋白表达可能是Galectin-3在CCl_4对肝细胞损伤中发挥保护作用的一个途径.     

白芍总苷对大鼠心肌缺血再灌注损伤保护作用及对GRP78表达的影响

目的研究白芍总苷(TGP)对心肌缺血再灌注损伤的保护作用及对Grp78蛋白表达的影响。方法采用传统的在体心肌缺血再灌注模型,观察TGP对大鼠心功能的影响。方法将48只大鼠随机分为6组:假手术组(sham组)、模型组(I/R)、TGP低(50mg/kg)组、TGP中(100mg/kg)组、TGP高(200mg/kg)组、葛根素(100mg/kg)组,分别在术前一周灌胃,假手术组和模型组分别给予等量溶媒,葛根素(PUE)于再灌注前5min经颈总静脉注射,分别测定缺血前、缺血后30min、再灌注90min时心率(HR)、左心室压力变化最大速率(±dp/dtmax)、左心室收缩峰压(LVSP)以评价心脏功能。结果缺血再灌注模型组在缺血30min及再灌注90min后LVSP和±dp/dtmax降低,与I/R组相比,不同剂量的TGP组与PUE组相对应时间点LVSP、±dp/dtmax进行性升高。结论TGP对心肌缺血再灌注损伤有保护作用;其机制可能与通过促进GRP78的表达来发挥内源性的保护作用有关。