高糖和脂多糖对腹膜间皮细胞中骨桥蛋白的表达及其所致纤维化的影响

目的观察高糖、脂多糖(LPS)单独及联合作用对大鼠腹膜间皮细胞(RPMC)中骨桥蛋白(OPN)的表达及其所致纤维化的影响。方法将培养的RPMC分为正常对照组,不同浓度葡萄糖组(1.5%,2.5%,4.25%,24 h),高糖(2.5%)作用不同时间组(8,12,24,34,48 h),不同浓度LPS组(50,100μg/mL),LPS(50μg/mL)作用不同时间组(8,12,24,34,48 h)及联合作用组(2.5%葡萄糖+LPS 50μg/mL,24 h)。采用细胞免疫组织化学法检测OPN在RPMC内的表达情况,Real-time RT-PCR技术检测OPN和转化生长因子β1(TGF-β1)mRNA的表达。结果高糖、LPS单独及联合作用均可刺激RPMC的OPN表达增加,呈剂量及时间依赖性,2者联合作用时OPN表达量较高糖、LPS单独作用时增高,差异有统计学意义(P<0.05)。高糖、LPS单独及联合作用均可导致TGF-β1表达增加,作用8 h时表达水平升高,12 h降低,24 h达到高峰,48 h仍存在,与0 h组比较,差异均有统计学意义(P<0.01)。结论高糖及LPS通过上调OPN的表达引起腹膜损伤,导致腹膜纤维化,2者联合作用时通过影响OPN的表达加速了腹膜透析合并腹膜炎过程中腹膜纤维化的病理进程。     

罗格列酮对高糖状态下人脐静脉内皮细胞血红素加氧酶-1表达的影响

目的:观察罗格列酮对高糖状态下人脐静脉内皮细胞(HUVECs)血红素加氧酶-1(HO-1)表达及细胞活力的影响。方法:体外培养HUVECs,分别在不同时间和用不同浓度的罗格列酮干预高糖状态下培养的HUVECs,提取细胞总RNA和蛋白,RT-PCR和Western blot检测HO-1表达的改变。采用台盼蓝染色法,检测细胞死亡率的变化。结果:与0μmol/L罗格列酮组及作用0h组相比,罗格列酮呈时间和浓度依赖性,提高高糖状态下HUVECs的HO-1表达,差别均具有统计学意义(P<0.05)。罗格列酮作用组HUVECs死亡率明显下降,差别均具有统计学意义(P<0.05)。结论:罗格列酮可以提高内皮细胞在高糖状态下抗氧化应激的能力,起到保护血管的作用。     

吡格列酮对高糖作用下大鼠腹膜间皮细胞增殖和TGF-β1表达的影响

目的 观察吡格列酮(PIO)对高糖作用下大鼠腹膜间皮细胞(RPMC)细胞增殖及转化生长因子β1(TGF-β1)表达的影响.方法 原代培养RPMC;并用MTT和ELISA法测定吡格列酮对高糖(2.5%葡萄糖)影响下RPMC增殖及其分泌TGF-β1的影响.结果 高糖能显著抑制RPMC的增殖(P<0.01).而5～20 μmol/L的吡格列酮能部分逆转高糖对RPMC增殖的抑制作用(P<0.01);高糖能显著增加RPMC表达TGF-β1(P<0.01),5～20 μmol/L的吡格列酮能明显减弱高糖上调TGF-β1的作用(P<0.01).结论 吡格列酮可能通过逆转高糖对RPMC的增殖抑制作用,抑制高糖上调RPMC表达纤维化因子TGF-β1的作用,进而修复高糖所致的RPMC的损伤,延缓腹膜纤维化,为改善间皮细胞损伤和防治腹膜纤维化提供实验依据.     

脂多糖对小鼠巨噬细胞TREM-1表达的影响

目的:初步探讨脂多糖(LPS)应激下小鼠巨噬细胞TREM-1的表达。方法:以小鼠巨噬细胞为研究对象,分别加入不同浓度的LPS(0.04、0.2、1、5和25μg/mL)作用后,采用RT-PCR法检测不同时间点LPS应激的巨噬细胞TREM-1mRNA的表达。结果:LPS呈时间依赖性地上调巨噬细胞TREM-1mRNA的表达,且在LPS作用6h达到峰值;LPS呈剂量依赖性地上调巨噬细胞TREM-1mRNA的表达,且在1μg/mL达到峰值。结论:LPS可上调小鼠巨噬细胞TREM-1mRNA的表达。     

从丹皮酚对TLR-NF-κB信号通路的影响探讨高血压病血瘀证的形成机制

【目的】通过观察活血化瘀中药牡丹皮的提取物丹皮酚(paeonol,Pae)对高血压病血瘀证患者血清损伤的血管内皮细胞TLR-NF-κB信号通路的影响,探讨高血压病血瘀证的形成机制。【方法】以体积分数10%的高血压病血瘀证、非血瘀证患者及健康人(正常组)血清干预人脐静脉内皮细胞(HUVEC-C)24 h,采用实时荧光定量(PCR)法检测Toll样受体4(TLR4)mRNA表达。以不同浓度的丹皮酚(大、中、小剂量分别为400μg/mL、200μg/mL、100μg/mL)干预脂多糖(LPS)诱导的HUVEC-C TLR4表达(24 h)及核转录因子(NF-κB)活化(2 h)。采用PCR法检测TLR4 mRNA及NF-κB mRNA的表达,免疫印迹(Western blot)技术检测TLR4蛋白及NF-κB蛋白(以I-κBα蛋白的降解反映)的表达。【结果】血清作用24 h后,TLR4 mRNA的表达按正常组、非血瘀证组、血瘀证组的顺序逐渐增高,组间差异有统计学意义(P0.01)。丹皮酚可呈剂量依赖性减少LPS诱导的TLR4 mRNA高表达(P0.001),抑制LPS诱导的TLR4蛋白高表达和I-κBα蛋白的降解。【结论】TLR-NF-κB信号途径介导的炎症反应和免疫紊乱是高血压病血瘀证形成的机制之一。     

几丁糖对高糖腹透液刺激大鼠腹膜间皮细胞TGF-β1表达的影响

目的 探讨高糖腹透液(4.25%)对大鼠二代腹膜间皮细胞TGF-β1表达的影响及几丁糖(chitosan)的干预作用.方法 取第2代鼠腹膜间皮细胞(rat peritoneal mesothelial cells, RPMC)(经细胞形态和免疫组化鉴定),随机分为4组(n=4),A组即对照组:DMEM/F12培养基培养;B组:DMEM/F12培养基中加入4.25%葡萄糖透析液;C组:DMEM/F12培养基中加入4.25%葡萄糖透析液+几丁糖1.2 mg/L;D组:DMEM/F12培养基中加入4.25%葡萄糖透析液+几丁糖0.4 mg/L.培养24 h收集细胞提取RNA,48 h收集细胞提取蛋白,RT-PCR检测腹膜间皮细胞TGF-β1 mRNA的表达.Western印迹检测TGF-β1的蛋白表达.结果 ①RPMC经高糖腹透液(4.25%)刺激后,TGF-β1 mRNA和蛋白的表达均上升,与0 h比较,TGF-β1 mRNA和蛋白的表达分别于24、48 h时达到高峰, A、B两组比较差异有统计学意义(P<0.05);②24 h时,几丁糖使TGF-β1 mRNA的表达下降明显(P<0.05),B组分别为A组的2.51倍、C组的2.13倍、D组的1.68倍,D组为C组的1.26倍且抑制程度与几丁糖浓度呈正相关(r=0.948, P<0.01).48 h时,几丁糖使TGF-β1的蛋白水平下降明显,B组分别为A组的1.85倍、C组的1.52倍、D组的1.24倍,D组为C组的1.23倍.C、D两组分别与B组比较差异有统计学意义(P<0.05),且抑制程度与几丁糖浓度呈正相关(r=0.831, P<0.05).结论 ①高糖腹透液可刺激RPMC TGF-β1 mRNA和蛋白表达上调;②几丁糖可抑制腹膜纤维化的中枢性因子TGF-β1表达的活性.     

Toll样受体4在人肺泡上皮细胞株中的表达及其在细胞炎症反应中的作用

目的 明确人Ⅱ型肺泡上皮细胞是否表达Toll样受体4(TLR4),探讨TLR4在慢性阻塞性肺疾病(COPD)气道炎症中的作用.方法 将人Ⅱ型肺泡上皮细胞系A549细胞分为正常对照组、转染腺病毒E1A质粒(E1A+组)和转染不含有腺病毒E1A的空白质粒组(E1A-组),分别以不同浓度的脂多糖(LPS,0、0.1、1、10μg/ml)、白细胞介素1β(IL-1β,0、0.1 ng/ml)、香烟提取物(CSE,0、10%、20%、40%)刺激.刺激后12、24 h,用反转录聚合酶链反应技术检测各组细胞IL-8 mRNA和TLR4 mRNA的表达,用蛋白质印迹技术测定核因子κB(NF-κB)亚单位P65蛋白的表达.结果 10μg/ml LPS、0.1 ng/ml IL-1β或20%CSE刺激24 h后,E1A+组IL-8 mRNA表达量(2.82、1.87、4.70)明显高于正常对照组(0.95、0.78、1.02,均P<0.05)和E1A-组(0.97、0.81、1.12,均P<0.05).10μg/ml的LPS刺激后12、24 h,E1A+组细胞TLR4 mRNA表达水平分别为4.52、7.99,明显高于正常对照组(1.91、3.81,均P<0.05)和E1A-组(2.00、3.88,均P<0.05);IL-1β也可增加TLR4 mRNA表达.CSE刺激对各组细胞TLR4 mRNA表达水平均无明显影响.3种刺激因子均可引起各组细胞NF-κB亚单位P65蛋白表达水平增高.结论 人Ⅱ型肺泡上皮细胞表达TLR4,LPS、IL-1β引起IL-8释放可能与TLR4介导的NF-κB激活有关.     

脂多糖对人单核细胞株THP-1细胞HIF-1α及其靶基因GLUT-1表达的影响

目的研究脂多糖(LPS)对人单核细胞株THP-1细胞低氧诱导因子-1α(HIF-1α)及其靶基因葡萄糖转运载体-1(GLUT-1)表达的影响。方法以1μg/mL LPS分别处理体外常规培养的THP-1细胞0、2、4、6、8 h。采用Western blotting检测THP-1细胞HIF-1α蛋白表达,RT-PCR技术检测HIF-1αmRNA与GLUT-1 mRNA表达。以不同浓度的LPS(0、0.01、0.1、1μg/mL)分别刺激THP-1细胞6 h,Western blotting检测HIF-1α蛋白表达。结果1μg/mL LPS作用2 h时,THP-1细胞的HIF-1α蛋白表达开始增加,4 h时显著增加(P<0.05),6~8 h时达高峰(P<0.01)。0.01μg/mL LPS刺激6 h,THP-1细胞的HIF-1α蛋白表达较对照组明显升高(P<0.05);当0.1μg/mL LPS和1μg/mL LPS刺激6 h时,HIF-1α蛋白表达更高,与对照组相比差异有高度统计学意义(P<0.01)。提示LPS能够诱导THP-1细胞内HIF-1α蛋白表达,并且呈时间和浓度依赖性。1μg/mL LPS作用2、4、6、8 h均能上调THP-1细胞HIF-1α与GLUT-1 mRNA水平(P<0.01),且随时间的延长表达逐渐增加。结论LPS能够促进人单核细胞株THP-1细胞HIF-1α及其靶基因GLUT-1的表达。     

大黄素对脂多糖诱导的角膜基质细胞表达细胞因子的影响

目的 观察大黄素对脂多糖(LPS)诱导的角膜基质细胞表达细胞因子的影响.方法 原代培养角膜基质细胞,选取第4代细胞进行实验.根据在LPS刺激角膜基质细胞前是否先用40μmol/L大黄素培养细胞30min,将细胞分为大黄素+LPS组和LPS组,分别在10μg/L LPS刺激角膜基质细胞前以及刺激后1、2、4、8 h收集细胞;或先用0、5、10、20和40 μmol/L的大黄素培养细胞30 min后,再用10μg/L LPS刺激8 h.Western blot检测角膜基质细胞κB抑制因子-α(IκB-α)蛋白的表达;逆转录聚合酶链反应(RT-PCR)检测角膜基质细胞白细胞介素(IL)-6和IL-8的mR-NA表达.结果 LPS刺激角膜基质细胞后1、2、4、8h,细胞表达IκB-α蛋白均较刺激前明显下降(P<0.01);大黄素对LPS引起的角膜基质细胞IκB-α蛋白的降解有不同程度的抑制作用,且该作用呈一定的浓度依赖性(P<0.05).LPS刺激引起角膜基质细胞IL-6和IL-8 mRNA表达升高,刺激后各时间点IL-6和IL-8 mRNA表达均较LPS刺激前明显升高,呈一定时间依赖性(P<0.01);大黄素对LPS诱导的角膜基质细胞IL-6和IL-8 mRNA表达有明显抑制作用,且该作用呈一定浓度依赖性(P<0.05).结论 大黄素能抑制体外培养的角膜基质细胞表达IL-6和IL-8,可能与其抑制IκB-α降解、促进NF-kB活化有关.     

罗格列酮对高糖培养的大鼠系膜细胞增生及p27表达的影响

目的:探讨罗格列酮对糖尿病肾病保护作用的机制。方法:体外培养的大鼠系膜细胞分为正常对照组(NG,含5.6mmol/L D-葡萄糖),NG+甘露醇(25 mmol/L)组,高糖组(HG,含30mmol/L D-葡萄糖),HG+罗格列酮(5μmol/L)组,HG+罗格列酮(10μmol/L)组,HG+罗格列酮(20μmol/L)组。用MTT法测定细胞增生,免疫细胞化学法测定p27Kip1表达。结果:(1)培养72小时后,在模拟高血糖的高糖培养基中,各浓度罗格列酮均可抑制大鼠系膜细胞增生,且此作用与渗透压改变无关。(2)与NG组相比,HG组培养24小时p27表达降低(P<0.05),与其增生活跃相一致,随时间延长,p27表达量逐渐增加,48、72小时无明显差异。与HG组相比,5μmol/L罗格列酮组作用24、48小时无明显差异,作用72小时p27表达增加(P<0.05);10μmol/L、20μmol/L罗格列酮组在各时间点均增加p27表达,有统计学意义。随着罗格列酮剂量增加,p27表达也明显增加。结论:在高糖环境下罗格列酮至少部分通过增加p27表达引起系膜细胞增生抑制,从而对糖尿病肾病起保护作用。     

Acupuncture Treatment of Vomiting

CASE HISTORY A female patient, 46 years old, head of the foreign affairs department of a certain university in Beijing, paid her first visit on October 9, 2006, with the chief complaint of vomiting for one month. She got vomiting after meals in early September. Before that, she had discomfortable sensation in the stomach due to angry with others, but she didn't pay much attention. Later, it developed into vomiting after eating. After the vomiting, the discomfort would be relieved, but with slight hypodynamia. She was once diagnosed as having 'neurogenic vomiting'. Having taken some western and Chinese drugs, the above symptoms were a little bit improved, but she would have nausea upon eating and with regurgitation. Because of the fear for vomiting, she did not dare to have food intake, with body weight reduction of 6 kilos in one month.     

Qiu Baoguo''''s Experience in Treating Consumptive Syndromes after Radio-Chemotherapies

Radiotherapy and chemotherapy are the important modern medical therapies for malignant tumors,yet they can also bring about serious local and systemic toxic side reactions so to decrease the patient;'s life quality,manifested by a series of consumptive symptoms.Having engaged in the combined work of Chinese and western medicine for nearly 50 years,the research fellow Qiu Baoguo in Henan Provincial Academy of TCM has developed his unique views on the TCM study of consumptive syndromes.The author of this essay had once the fortune tO follow Dr.Qiu in clinic,and specially would like to introduce in the following Dr.Qiu's experience in treating consumptive syndromes after radio-chemotherapies for patients with malignant tumor.     

Insomnia Due to Deficiency of Both the Heart and Spleen Treated by Acupuncture-Moxibustion and Chinese Tuina

OBJECTIVE: To observe therapeutic effects of the comprehensive therapy of acupuncture-moxibustion and Chinese Tuina for treatment of insomnia due to deficiency of both the heart and spleen. METHODS: 92 cases were divided randomly into the treatment group (treated by acupuncture-moxibustion and Chinese Tuina) and the control group (treated by acupuncture-moxibustion). RESULTS: The therapeutic effect of the treatment group was obviously superior to that of the control group (the CHI2 test showed P < 0.01). CONCLUSIONS: The comprehensive therapy of acupuncture-moxibustion and Chinese Tuina can give marked therapeutic effects for treatment of insomnia due to deficiency of both the heart and spleen.     

Analysis of CK-19,PDX-1,Nestin,Ngn3 in different donor islet purity in rats

Objective: To investigate if there are the CK-19, PDX-1, Nestin, Ngn3 positive cells in the donor islets of different purity in rats. Methods: Thirty male adult SD rats were randomly divided into 3 groups. Islets were isolated using digestion by ductal injection of collagenase. Group Ⅰ (n=10): Separating cell preparations were not purified, Group Ⅱ(n=10): Islet sediment was purified with 25% Ficoll400 ,Group Ⅲ (n=10): Islet sediment was purified with 25% and 11% Ficoll-400. The levels of protein of CK-19, PDX-1, Nestin and Ngn3 were detected by immunohistochemistry and the mRNA of CK-19, PDX-1, Nestin, Ngn3 was amplified by RT-PCR. Results: After two different purification methods applied, three islet preparations of different purities were obtained. The difference of islet purity was significant among various groups (P＜0.05). Compared with group Ⅱ and group Ⅲ,the protein and mRNA of CK-19, PDX-1, Nestin,Ngn3 were both higher in group Ⅰ; group Ⅲ was poorly expressed. Conclusions: The three different islet purity donor islet have different CK-19, PDX-1, Nestin, Ngn3 positive cells within them, indicating that there are some islet stem cells in the purified donor islet.     

A preliminary screening study on the associated proteins in human psoriasis vulgaris by serum proteomics technologies

Objective：To investigate the optimum screening conditions of associated proteins in human psoriasis vulgaris by serum proteomics technique, and to screen the different expression proteins related with psoriasis vulgaris. Methods：Serum samples of peripheral blood were collected from newly diagnosed psoriasis vulgaris patients in the clinic, and 20 matched healthy persons. Serum albumin IgG was removed by filtering with ProteoExtract Albumin/IgG. After comparative proteomics analysis the different protein spots were identified using 2-DE and MS. Results ：Electrophoresis figures with high resolution and reproducibility were obtained. Three different expression proteins were found only in the serum from psoriasis vulgaris patients, while nine other different proteins expressing from healthy volunteers. Conclusion：The protein expression was different in the serum between the psoriasis vulgaris patients and healthy volunteers. It was hoped that we could find the biomarkers related to psoriasis vulgaris by using proteomics.     

A CASE OF HUGE GASTRIC DISOPYROBEZOAR

PYTOBEZOARS are common bezoars in gastrontestinal tract, commonly seen in stomach and mall intestine,1but huge disopyrobezoars are     

Acupuncture Treatment of Chest Bi Syndrome

CASE HISTORY A male,a retired officer of 61 years old,paid his first visit on May 12,2006.The patient stated that 2 weeks before he suddenly got paroxysmal precordial choking pain,with irregular attacks.Each attack would last several seconds to several minutes.The attack was accompanied with heavy oppressed sensation in the precordial region,palpitation,lassitude,and in severe cases,sweating,which was once diagnosed by a hospital as'coronary heart disease'.Recalling the case history,the patient said that several days before he got angry with others because of some affairs in work,and he was not so used to the life pattern since he had just retired from his busy working post.At usual times,he was quick tempered and peevish.In recent days,he had poor appetite,and loose stool.     

Instructions to Contributors

The English edition of Chinese Journal of Integrative Medicine is edited and published by the Chinese Journal of Integrated Traditional and Western Medicine Press.Original scientific papers of advanced clinical and experimental medicine in the field of complementary/alternative medicine or integrative medicine are expected.     

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