颗粒体上皮蛋白前体在脓毒血症中的变化及其与疾病严重程度的关系

目的探讨血清颗粒体上皮蛋白前体(progranulin,PGRN)在脓毒血症中的变化及其与炎症反应和疾病严重程度的关系。方法选择急诊重症监护病房(ICU)确诊脓毒血症的41例患者为研究对象,并以20例健康志愿者作为对照组。采用酶联免疫吸附试验(ELISA)法分别检测入ICU当天、治疗3及7 d血清PGRN浓度,并测定炎症指标降钙素原(PCT)、C反应蛋白(CRP)和肿瘤坏死因子-α(TNF-α)水平,并记录入院时急性生理学与慢性健康状况评分系统Ⅱ(APACHEⅡ)、序贯器官衰竭评分(SOFA),以及住院时间和机械通气时间,并分析血清PGRN水平与上述指标的相关性。结果脓毒症患者收入ICU当天血清PGRN浓度较对照组显著下降(P<0.05),入院治疗3 d及7 d后,血清PGRN浓度与入ICU当天相比较逐步上升(P<0.05)。入ICU时血清PGRN浓度与炎症因子PCT(r=-0.615,P<0.05)、CRP(r=-0.635,P<0.01)、TNF-α(r=-0.600,P<0.05)浓度均负相关,且与APACHEⅡ(r=-0.663,P<0.01)、SOFA评分(r=-0.590,P<0.05)呈显著负相关。结论脓毒症患者血清PGRN浓度较正常人明显降低,且与炎症水平及脓毒症严重程度有关,提示PGRN可能作为一种保护性蛋白在脓毒血症发病过程中发挥保护作用,因此,监测脓毒血症患者血清PGRN变化趋势对判断脓毒血症炎症程度和预后一定的临床意义。

脓毒症患儿血SAA、sTREM-1、NLR、PGRN的变化及预后因素分析

目的分析脓毒症患儿淀粉样蛋白A(SAA)、可溶性髓样细胞触发受体-1(sTREM-1)、中性粒细胞与淋巴细胞比值(NLR)、颗粒体上皮蛋白前体(PGRN)变化,分析相关预后影响因素。方法选取2019年1月至2022年12月河北中石油中心医院收入的脓毒血症患儿120例为观察组,另匹配同时间段非脓毒症健康新生儿46例为健康组。根据观察组患儿28d存活、死亡情况分为存活组(94例)、死亡或放弃治疗组(26例),比较脓毒症患儿、健康新生儿SAA、sTREM-1、NLR、PGRN水平。采用二元Logistic回归分析影响脓毒症患儿预后的因素,建立受试者工作特征(ROC)曲线,评估SAA、sTREM-1、NLR、PGRN预测发展结局的价值。结果与健康组相比,观察组患儿SAA、sTREM-1、NLR水平更高,PGRN水平更低,差异有统计学意义(t值分别为44.598、7.137、7.141、-16.695,P<0.05);与存活组相比,死亡或放弃治疗组患儿入院心率、入院呼吸频率、胎龄≤28周占比、SAA、sTREM-1、NLR水平更高,PGRN水平、入院氧饱和度更低,差异有统计学意义(χ^(2)/t值介于-5.388~9.038之间,P<0.05);多因素Logistic回归分析结果显示入院心率、SAA、sTREM-1、NLR是影响脓毒症患儿预后的危险因素,其OR值及95%CI分别为1.534(1.203~1.955)、1.037(1.017~1.058)、1.255(1.052~1.496)、1.755(1.228~2.507),氧饱和度、PGRN是影响脓毒症患儿预后的保护因素,其OR值及95%CI分别为0.282(0.142~0.560)、0.926(0.884~0.970);SAA、sTREM-1、NLR、PGRN预测脓毒症患儿发展结局的曲线下面积(AUC)分别为0.804、0.718、0.873、0.780,联合预测的曲线下面积为0.928。结论脓毒症患儿血SAA、sTREM-1、NLR较健康新生儿异常升高,是影响患儿预后的危险因素;PGRN水平异常降低,是影响患儿预后的保护因素。

人食管和贲门癌前病变和癌组织P40上皮特异性蛋白表达的研究

利用鼠单克隆抗体７Ｅ（１２）Ｈ（１２），对人食管和贲门正常粘膜上皮，癌前病变和癌组织的Ｐ４０上皮特异性蛋白表达进行比较研究。结果表明，食管和贲门上皮Ｐ４０蛋白的表达和分布明显不同。贲门柱状上皮Ｐ４０蛋白的免疫反应强度和发生率明显高于鳞状上皮（相差８倍）。食管固有腺上皮细胞也出现较强的Ｐ４０蛋白免疫反应性。在贲门组织中，Ｐ４０蛋白免疫反应６２％发生在细胞浆；２１％发生在细胞核；１７％同时发生在胞浆和胞核。而在食管，其反应全部在胞浆。随着病变的加重，食管和贲门Ｐ４０蛋白的表达均出现增高的趋势。

EPLIN基因在恶性肿瘤发生发展中的作用研究进展

肿瘤中丢失的上皮蛋白(EPLIN)是一种参与调节肌动蛋白细胞骨架的分子,其在肿瘤中表现为频繁下调或丢失,与乳腺癌、前列腺癌、食管癌和胃癌等实体瘤的进展和转移相关,可通过各种机制参与恶性肿瘤的发生发展。过表达EPLIN是一种有效的治疗手段,可抑制细胞生长和运动,降低细胞侵袭性,有望成为肿瘤治疗的潜在预后生物标志物和治疗靶点。未来深入研究EPLIN基因在恶性肿瘤中的作用机制,可以为恶性肿瘤的早期诊断和靶向治疗提供新思路。

血清PTX3、HBP、PGRN水平与产褥期细菌感染所致脓毒症的关联性分析

目的:探讨血清正五聚体蛋白3(PTX3)、肝素结合蛋白(HBP)、颗粒体上皮蛋白前体(PGRN)与产褥期细菌感染所致脓毒症的关联性及对病情的评估价值。方法:选取某院2020年4月—2022年4月诊治的产褥期细菌感染所致脓毒症患者154例为研究组,选取同期产褥期健康产妇154例为对照组,比较2组入院时血清PTX3、HBP、PGRN水平。根据病情程度将研究组分为普通脓毒症组(48例)、严重脓毒症组(62例)、脓毒症休克组(44例)3个亚组,比较3组入院时血清PTX3、HBP、PGRN水平及急性生理学与慢性健康状况(APACHEⅡ)评分、序贯器官衰竭(SOFA)评分,采用Pearson分析入院时血清PTX3、HBP、PGRN水平与APACHEⅡ、SOFA评分相关性。随访3个月后,比较不同病情转归患者入院2周后血清PTX3、HBP、PGRN水平,危险度分析其对脓毒症预后的预测价值。结果:入院时,研究组血清PTX3、HBP水平高于对照组,PGRN水平低于对照组,差异均有统计学意义(P<0.05)。3组不同病情程度患者血清PTX3、HBP水平及APACHEⅡ、SOFA评分比较:普通脓毒症组<严重脓毒症组<脓毒症休克组;血清PGRN水平比较:普通脓毒症组>严重脓毒症组>脓毒症休克组,差异均有统计学意义(P<0.05)。Pearson相关性分析显示,入院时血清PTX3、HBP水平与APACHEⅡ、SOFA评分呈正相关,PGRN水平与APACHEⅡ、SOFA评分呈负相关(P<0.05)。入院2周后,存活组血清PTX3、HBP水平低于病死组,PGRN水平高于病死组,差异均有统计学意义(P<0.05)。危险度分析显示,血清PTX3、HBP、PGRN高水平组入院3个月内病死风险分别为低水平组的4.362、3.047、0.249倍(P<0.05)。结论:血清PTX3、HBP、PGRN水平与产褥期细菌感染所致脓毒症的发生、发展密切相关,临床可通过其评估病情及预测转归结局,以针对性制定干预方案,改善预后。

氦氖激光穴位照射对缺血缺氧后新生大鼠海马神经元增殖的影响

目的:探讨氦氖激光穴位照射疗法对缺血缺氧新生大鼠脑损伤后海马神经元增殖的影响。方法:7d龄健康Wistar大鼠72只随机分成4组:假手术组、缺血缺氧组、缺血缺氧激光穴位照射组、缺血缺氧激光非穴位照射组。制备新生大鼠缺血缺氧脑损伤模型,激光穴位照射组选择“百会”和“大椎”穴位给予氦氖激光照射,激光非穴位照射组选择腹部非穴位部位进行激光照射,常规饲养22d后处死,处死前经腹腔注射5溴2脱氧脲嘧啶(BrdU)标记增殖细胞。取左侧脑做组织切片,HE、Nissl染色以及采用抗BrdU抗体和神经上皮蛋白(Nestin)抗体进行免疫组织化学染色。结果:激光穴位照射组与其他组比较海马神经元胞体内尼氏体丢失较少,神经元坏死减轻,海马齿状回BrdU标记的免疫阳性细胞数增加,海马CA1区Nestin免疫阳性细胞数增高。结论:激光穴位照射对海马神经元有保护作用,促进了海马神经元的增殖。

BIGH3基因与人角膜营养不良

BIGH3基因突变有关的角膜营养不良现今已是角膜营养不良最常见的遗传学致病因素。在临床上,BIGH3基因突变主要导致两大类角膜营养不良,一类是颗粒状角膜营养不良,另一类是格子状角膜营养不良。阅读了大量关于BIGH3基因导致的角膜营养不良文献,现从BIGH3的发现、表达产物、基因突变位点、导致临床角膜营养不良的分型及可能致病原因做一详细综述。

Progranulin在糖脂代谢及肾脏疾病调控机制中的研究进展

颗粒体上皮蛋白前体(PGRN)广泛分布于上皮细胞、免疫细胞、神经细胞等多种细胞中,具有多种功能。完整形式的PGRN可通过与肿瘤坏死因子α(TNF-α)结合,进而抑制TNF-α/TNF受体通路来发挥抗炎作用,而经蛋白酶加工后的PGRN通过增加促炎因子的产生而具有促炎效应。PGRN可作为抗炎因子参与早期胚胎发育和组织重塑,还可通过抑制炎症、促进自噬在肾脏疾病发生发展过程中发挥重要作用,此外,PGRN也可作为脂肪细胞因子参与肥胖和胰岛素抵抗等过程,参与肥胖和糖尿病的发生。

血清PGRN及PCT的表达水平与脓毒症患儿预后的关系

目的:探讨颗粒体上皮蛋白前体(PGRN)及降钙素原(PCT)在血清中的表达水平与脓毒症患儿预后的关系。方法:选取2019年1月-2020年12月本院儿科治疗的251例脓毒症患儿,根据28 d随访是否存活,将所有患儿分为死亡组68例和存活组183例。通过小儿危重病例评分(PCIS)评估患儿病情程度;通过酶联免疫吸附法检测血清中PGRN及PCT的表达水平。采用Pearson相关考察血清PGRN、PCT与PCIS评分间关系;多因素Logistic回归确定预后危险因素;并通过绘制ROC曲线分析PGRN、PCT对患儿预后的预测价值。结果:与存活组患儿比较,死亡组具有更低的PCIS评分和血清PGRN水平,而血清PCT水平更高,差异均有统计学意义(P<0.05);Pearson相关显示,血清PGRN与脓毒症患儿的PCIS评分呈正相关(r=0.525,P<0.001),而血清PCT与其PCIS评分呈负相关(r=-0.536,P<0.001)。多因素Logistic分析结果可知,PCIS评分(OR=2.348,95%CI:1.252,4.403)、血清PGRN(OR=2.410,95%CI:1.287,4.514)及血清PCT(OR=2.702,95%CI:1.350,5.406)均是影响脓毒症患儿28 d死亡的独立危险因素(P<0.05)。ROC曲线结果显示,曲线下面积及约登指数PCT<PGRN<联合预测。二者联合检测的约登指数为0.65,敏感度为0.84,特异度为0.81,曲线下面积为0.905。结论:血清PGRN、PCT水平与脓毒症患儿PCIS评分相关,二者均可作为其预后预测的潜在标志物。

Epithelial-mesenchymal transition mediated tumourigenesis in the gastrointestinal tract

Epithelial-mesenchymal transition (EMT) is a highly conserved process that has been well characterised in embryogenesis. Studies have shown that the aberrant activation of EMT in adult epithelia can promote tumour metastasis by repressing cell adhesion molecules,including epithelial (E)-cadherin. Reduced intracellular adhesion may allow tumour cells to disseminate and spread throughout the body. A number of transcription proteins of the Snail superfamily have been implicated in EMT. These proteins have been shown to be over-expressed in advanced gastrointestinal (GI) tumours including oesophageal adenocarcinomas,colorectal carcinomas,gastric and pancreatic cancers,with a concomitant reduction in the expression of E-cadherin. Regulators of EMT may provide novel clinical targets to detect GI cancers early,so that cancers previously associated with a poor prognosis such as pancreatic cancer can be diagnosed before they become inoperable. Furthermore,pharmacological therapies designed to inhibit these proteins will aim to prevent local and distant tumour invasion.

Effect of recombinant human interleukin-11 on expressions of interleukin-11 receptorα-chain and glycoprotein 130 in intestinal epithelium cell line-6 after neutron irradiation

AIM： To explore the effect of recombinant human interleukin-11 （rhIL-11） on the expressions of interleukin-11 receptor α-chain （IL-11Rα） and an additional signal transducer glycoprotein 130 （gp130） in intestinal epithelium cell line-6 （IEC-6） after neutron irradiation. METHODS： Cultured IEC-6 cells were exposed to 4.0Gy neutron and treated with 100 ng/mL rhIL-11 12 h prior to or immediately after irradiation. The apoptosis and necrosis rates and expressions of IL-11Rα and gp130 were observed by flow cytometry, immunohistochemistry, Western blot and image analysis. RESULTS： The apoptosis rate of IEC-6 cells was increased by irradiation at 6 h （P 〈 0.01）, IL-11 stimulation resulted in a decreased apoptosis rate in irradiated IEC-6 cells （P 〈 0.05）. In normal control IEC-6 cells, intense immunoreactivity of IL-11Rα was located within the cell membrane and cytoplasm. The level of IL-11Rα expression significantly decreased at 6 h after irradiation （P 〈 0.01） and restored at 24 h after irradiation. In IEC-6 cells treated with both radiation and rhIL-11, the level of IL- 11Rα expression was higher than that of irradiated cells （P 〈 0.05）. When it came to gp130 protein, it was located in the cytoplasm of IEC-6 cells. After irradiation, we found a progressive decrease in the expression of gp130 protein （P 〈 0.05） in 48 hours post-radiation, while in rhIL-11-stimulated cells, it came back to normal level at 24 h after irradiation and decreased at 48 h, but was still higher than that of only irradiated cells （P 〈 0.05）. CONCLUSION： rhIL-11 can protect IEC-6 cells from neutron irradiation. The protective effect of rhIL-11 might be connected with its ability to up-regulate the expressions of specific ligand-binding subunit IL-11Rα and signal-transducing subunit gp130.

Roles of CREB-binding protein (CBP)/p300 in respiratory epithelium tumorigenesis

CREB-binding protein （CBP） and its homologue p300 are transcriptional co-activators of various sequence-specific transcription factors that are involved in a wide array of cellular activities, such as DNA repair, cell growth, differentia- tion and apoptosis. Several studies have suggested that CBP and p300 might be considered as tumour suppressors, with their prominent role being the cross-coupling of distinct gene expression patterns in response to various stimuli. They exert their actions mainly via acetylation of histones and other regulatory proteins （e.g. p53）. A major paradox in CBP/ p300 function is that they seem capable of contributing to various opposed cellular processes. Respiratory epithelium tumorigenesis represents a complex process of multi-step accumulations of a gamut of genetic and epigenetic aberrations. Transcription modulation through the alternate formation of activating and repressive complexes is the ultimate converging point of these derangements, and CBP/p300 represents key participants in this interplay. Thus, illumination of their molecular actions and interactions could reveal new potential targets for pharmacological interventions in respiratory epithelium carcinogenesis.

Expression of SNC73, a transcript of the immunoglobulin α-1 gene, in human epithelial carcinomas

AIM： To investigate the expression of SNC73, a transcript of the immunoglobulin α-1 gene （IgA1-H chain）, in human epitheliα-derived tumor cells. METHODS： Total RNAs and cell lysates were prepared from five different human epithelial cell lines derived from lung, stomach, liver, skin, and breast, respectively. RT-PCR and immunoblot analysis of these five cell lines were done. Both RT-PCR and immunochemistry were used to detect the expression of SNC73 in these cell lines. We also examined the expression of SNC73 in normal epithelial cells of colon mucosa by in situ hybridization. RT-PCR and immunoblot analysis were used to determine whether the recombination activating gene1/2 （RAG1 and RAG2） is present. The expression of three immunoglobulin transcription factors, EBF, E2A and Pax5, and the heavy chain of IgA1 and two types of light chains of immunoglobulin （κ and λ） in the aforementioned cell lines were analyzed by RT-PCR and immunochemistry, respectively. All the RT-PCR products were analyzed by sequencing. RESULTS： The results of RT-PCR and immunochemistry showed that both mRNA and protein of SNC73 were expressed in five human epitheliα-derived cancer cell lines. These data were further confirmed in the normal epithelial cells of colon mucosa by in situ hybridization. Also, the heavy chain of IgA1 and κ light chain were detected in these cells, but no λ light chain was observed. Both RAG1 and RAG2 were expressed in these human epitheliα-derived cancer cell lines and the sequence was identical to that expressed in pre-B and pre-T cells. In addition to RAG1 and RAG2, the mRNA in one of the immunoglobulin transcription factors, EBF, was also detected in these cell lines, and Pax5 was only expressed in SW480 cells, but no expression of E2A was observed in all the five cell lines. CONCLUSION： Immunoglobulin A1 is originally expressed and V（D）J recombination machine is also present in non-lymphoid cells, suggesting that V（D）J recombination machine mediates the assembly of immunoglobulin A1 in non-lymphoid cells as in prelymphocytes.

Epithelial-to-mesenchymal and mesenchymal-to-epithelial transitions in the colon

Epithelial-to-mesenchymal and mesenchymal-to-epi- thelial transitions are well established biological events which have an important role in not just normal tissue and organ development, but in the pathogenesis of diseases. Increasing evidence has established their presence in the human colon during colorectal carcinogenesis and cancer invasion, chronic inflammation-related fibrosis and in the course of mucosal healing. A large body of evidence supports the role for transforming growth factor-13 and its downstream Smad signaling, the phosphatidylinositol 3＇-kinase/Akt/mTOR axis, the Ras-mitogen-activated protein kinase/Snail/Slug and FOXC2 pathway, and Hedgehog signaling and microR- NAs in the development of colorectal cancers via epi- thelial-to-mesenchymal transition. C-met and Frizzled-7, among others, seem to be the principle effectors of mesenchymal-to-epithelial transition, hence have a role not just in mucosal regeneration but in the progression of colonic wall fibrosis. Here we discuss a role for these pathways in the initiation and development of the transition events. A better understanding of their induction and regulation may lead to the identification of pathways and factors that could be potent therapeu- tic targets. The inhibition of epithelial-to-mesenchymal transition using mTOR kinase inhibitors targeting theATP binding pocket and which inhibit both mTORC1 and mTORC2, RNA aptamers or peptide mimetics, such as a Wnt5A-mimetic, may all be useful in both cancer treatment and delaying fibrosis, while the induction of mesenchymal-to-epithelial transition in induced pluripotent stem cells may enhance epithelial healing in the case of severe mucosal damage. The preliminary results of the current studies are promising, but more clinical investigations are needed to develop new and safe therapeutic strategies for diseases of the colon.

Inhibition of PMA-induced endothelial cell activation and adhesion by over-expression of domain negative IκBα protein

AIM: NF-κB, regulate the expression of cytokine-inducible genes involving immune and inflammatory responses, will be potential therapy approach for allograft from rejection. In this study, we use pCMV-IκBαM vector to inhibit NF-κB activation and investigate the effect of pCMV-IκBαM in inhibition of T cells adhesion to endothelial cells. METHODS: The NF-κB activity was detected with pNF-κB reporter gene and electrophoretic mobility shift assay. Expression of cell surface molecules was detected by RT-PCR and flow cytometer. The cell-cell adhesion assay was performed to determine the effect of pCMV-IκBαM in inhibition of T cells adhesion to endothelial cells. RESULTS: We could find that NF-κB activity is inhibited by over-expression of non-degraded IκBα protein. Expression of adhesion molecules like ICAM-1, VCAM-1, and P-selectin as well as cell-cell adhesion were inhibited significantly by transfection of the pCMV-IκBαM vector. CONCLUSION: Our results indicate that the pCMVIκBαM, which inhibit the activity of NF-κB through over-expression of non-degraded IκBα protein, can be used for gene therapy in diseases involving NF-κB activation abnormally like organ transplantation via inhibiting cell adhesion.

Epigenetic Repression of SATB1 by Polycomb Group Protein EZH2 in Epithelial Cells

Objective To study the regulatory mechanism of SATB1 repression in cells other than T cells or erythroid cells, which have high expression level of SATB1. Methods HeLa epithelial cells were treated with either histone deacetylase inhibitor (HDACi) trichostatin A (TSA) or DNA methylation inhibitor 5-Aza-C before detecting SATB1 expression. Luciferase reporter system was applied to measure effects of EZH2 on SATB1 promoter activity. Over-expression or knockdown of EZH2 and subsequent quantitative reverse transcription-polymerase chain reaction were performed to determine the effect of this Polycomb group protein on SATB1 transcription. Chromatin immunoprecipitation (ChIP) assay was applied to measure enrichment of EZH2 and trimethylated H3K27 (H3K27me3) at SATB1 promoter in HeLa cells. K562 cells and Jurkat cells, both having high-level expression of SATB1, were used in the ChIP experiment as controls. Results Both TSA and 5-Aza-C increased SATB1 expression in HeLa cells. Over-expression of EZH2 reduced promoter activity as well as the mRNA level of SATB1, while knockdown of EZH2 apparently enhanced SATB1 expression in HeLa cells but not in K562 cells and Jurkat cells. ChIP assay results suggested that epigenetic silencing of SATB1 by EZH2 in HeLa cells was mediated by trimethylation modification of H3K27. In contrast, enrichment of EZH2 and H3K27me3 was not detected within proximal promoter region of SATB1 in either K562 or Jurkat cells. Conclusion SATB1 is a bona fide EZH2 target gene in HeLa cells and the repression of SATB1 by EZH2 may be mediated by trimethylation modification on H3K27.

Effects of HLEC on the secreted proteins of epithelial ovarian cancer cells prone to metastasize to lymph nodes

Objective： To study explores the effect of HLEC on the secreted proteins of epithelial ovarian cancer （EOC） cells （SKOV3-PM4） with directional highly lymphatic metastasis. Methods： Supernatants of four groups of cultured cells, namely, SKOV3 （A）, SKOV3＋HLEC （B）, SKOV3-PM4 （C）, SKOV3-PM4＋HLEC （D）, were collected, and their proteins were detected by antibody arrays and iTRAOcZD-LC-MALDI- TOF/TOF/MS. Significantly differential proteins were further analyzed via bioinformatics and validated in human serums and cell media via ELISA. Results： Results of antibody arrays and mass spectrometry demonstrated that GRN and VEGFA were upregulated in group C （compared with group A）, whereas IGFBP7 and SPARC were downregulated in group D （compared with group C）. Comprehensive bioinformatics analysis results showed that IGFBP7 and VEGFA were closely linked to each other. Further validation with serums showed statistical significance in VEGFA and IGFBP7 levels among groups of patients with ovarian cancers, benign tumors, and control groups. Two proteins were upegulated in the first group. VEGFA in the control group was downregulated. For IGFBP, upregulation in the control group and down-regulation in the first group were also observed. Conclusion： The HLEC microenvironment is closely associated with directional metastasis to lymph nodes and with differential proteins including cell stromal proteins and adhesion factors. The upregulation of VEGFA and GRN and the downregulation of SPARC and IGFBP7 are closely associated with directional metastasis to lymph nodes in EOC cells.

A panel of monoclonal antibodies against the prion protein proves that there is no prion protein in human pancreatic ductal epithelial cells

Prion diseases are a group of neurodegenerative diseases that are fatal. The study of these unique diseases in China is hampered by a lack of resources. Amongst the most important resources for biological study are monoclonal antibodies. Here, we characterize a panel of monoclonal antibodies specific for cellular prion protein by enzyme-linked immunosorbent assay(ELISA), immunofluorescent staining, flow cytometry, and western blotting. We identify several antibodies that can be used for specific applications and we demonstrate that there is no prion protein expression in human pancreatic ductal epithelial cells(HPDC).

Effect of salvianolate on intestinal epithelium tight junction protein zonula occludens protein 1 in cirrhotic rats

AIM:To study the effect of salvianolate on tight junctions(TJs) and zonula occludens protein 1(ZO-1) in small intestinal mucosa of cirrhotic rats.METHODS:Cirrhosis was induced using carbon tetrachloride.Rats were randomly divided into the untreated group,low-dose salvianolate(12 mg/kg) treatment group,medium-dose salvianolate(24 mg/kg) treatment group,and high-dose salvianolate(48 mg/kg) treatment group,and were treated for 2 wk.Another 10 healthy rats served as the normal control group.Histological changes in liver tissue samples were observed under a light microscope.We evaluated morphologic indices of ileal mucosa including intestinal villi width and thickness of mucosa and intestinal wall using a pathological image analysis system.Ultrastructural changes in small intestinal mucosa were investigated in the five groups using transmission electron microscopy.The changes in ZO-1 expression,a tight junction protein,were analyzed by immunocytochemistry.The staining index was calculated as the product of the staining intensity score and the proportion of positive cells.RESULTS:In the untreated group,hepatocytes showed a disordered arrangement,fatty degeneration was extensive,swelling was obvious,and disorganized lobules were divided by collagen fibers in hepatic tissue,which were partly improved in the salvianolate treated groups.In the untreated group,abundant lymphocytes infiltrated the fibrous tissue with proliferation of bile ducts,and collagen fibers gradually decreased and damaged hepatic lobules were partly repaired following salvianolate treatment.Compared with the untreated group,no differences in intestinal villi width between the five groups were observed.The villi height as well as mucosa and intestinal wall thickness gradually thickened with salvianolate treatment and were significantly shorter in the untreated group compared with those in the salvianolate treatment groups and normal group(P < 0.01).The number of microvilli decreased and showed irregular lengths and arrangements in the untreated group.The intercellular space between epithelial cells was wider.The TJs were discontinuous,which indicated disruption in TJ morphology in the untreated group.In the treated groups,the microvilli in the intestinal epithelium were regular and the TJs were gradually integrated and distinct.The expression of ZO-1 decreased in the small intestine of the untreated cirrhotic rats.The high expression rate of ZO-1 in ileal mucosa in the untreated group was significantly lower than that in the medium-dose salvianolate group(21.43% vs 64.29%,χ 2 = 5.25,P < 0.05),high-dose salvianolate group(21.43% vs 76.92%,χ 2 = 8.315,P < 0.01) and normal group(21.43% vs 90%,χ 2 = 10.98,P < 0.01).CONCLUSION:Salvianolate improves liver histopathological changes,repairs intestinal mucosa and TJ structure,and enhances ZO-1 expression in the small intestinal mucosa in cirrhotic rats.

Inhibition of p38 mitogen-activated protein kinase may decrease intestinal epithelial cell apoptosis and improve intestinal epithelial barrier function after ischemia- reperf usion injury

AIM: To investigate the role of p38 mitogen-activated protein kinase in rat small intestine after ischemia-reperfusion (I/R)insult and the relationship between activation of p38 MAPK and apoptotic cell death of intestine.METHODS: Ninety Wistar rats were divided randomly into three groups, namely sham-operated group (C), I/R vehicle group (R) and SB203580 pre-treated group(S).In groups R and S, the superior mesenteric artery(SMA)was separated and occluded for 45 min, then released for reperfusion for0.25, 0.5, 1, 2, 6, 12 and 24 h. In group C, SMA was separated without occlusion. Plasma D-lactate levels were examined and histological changes were observed under a light microscope. The activity of p38 MAPK was determined by Western immunoblotting and apoptotic cells were detected by the terminal deoxynucleotidyl transferase (TdT)-mediated dUDP-biotin nick end labeling (TUNEL).RESULTS: Intestinal ischemia followed by reperfusion activated p38 MAPK, and the maximal level of activation (7.3-fold vs sham-operated group) was reached 30 min after I/R. Treatment with SB 203580, a p38 MAPK inhibitor,reduced intestinal apoptosis (26.72±3.39% vs62.50±3.08%in I/R vehicle, P＜0.01) and decreased plasma D-lactate level (0.78±0.15 mmol/L in I/R vehicle vs0.42±0.17 mmol/L in SB-treated group) and improved post-ischemic intestinal histological damage.CONCLUSION: p38 MAPK plays a crucial role in the signal transduction pathway mediating post-ischemic intestinal apoptosis, and inhibition of p38 MAPK may attenuate ischemia-reperfusion injury.