骨髓间充质干细胞移植对心肌梗死大鼠左心功能的影响

目的研究不同途径和不同浓度的骨髓间充质干细胞(MSCs)移植对心肌梗死大鼠左心功能的影响。方法结扎左冠状动脉前降支制备大鼠心肌梗死模型,采用同种异体大鼠MSCs体外分离、纯化、扩增,4’,6-二脒-2-苯基吲哚(DAPI)标记,将48只SD大鼠随机分为6组,A、B、C和D组于心肌梗死后2周经尾静脉移植MSCs(A组:细胞数5×105、B组:细胞数1×106、C组:细胞数5×106、D组:细胞数1×107),E组:经心外膜注射MSCs(细胞数5×106),对照组:注入等量培养基。4周后测定左心功能并行免疫组织化学检测。结果(1)经静脉和心外膜注射移植MSCs大鼠心肌组织中均可以观察到DAPI标记细胞存在,免疫组织化学阳性标记细胞α-肌动蛋白检测阳性,与对照组比较,两组左心功能均明显改善(P<0.05)和梗死面积均明显缩小[分别为(45±2)%,(40±4)%和(41±3)%,P<0.05]。(2)与对照组比较,A组左心功能无明显改善(P>0.05),B、C、D组左心功能明显改善(P<0.05),C和D组优于B组(P<0.05)。结论MSCs经不同途径归巢至心肌梗死区,改善心功能。     

同种异体移植骨髓间充质干细胞治疗大鼠心肌梗死

目的　探讨同种异体骨髓间充质干细胞 (MSCs)在梗死大鼠心脏局部存活、迁移、分化及对心功能的影响 ;明确同种异体细胞移植治疗心肌梗死 (MI)的可行性及效果。方法　雌性Wistar大鼠 3 5只 ,随机分为正常对照组、急性心肌梗死 (AMI)组及MI MSCs治疗组。分离纯化雄性Wistar大鼠骨髓MSCs ,于左冠状动脉前降支结扎后 1～ 3h植入到雌性大鼠心脏组织 ,移植后 10周检测心功能并取心脏检测各种相关指标。结果　异体大鼠MSCs经纯化后可在梗死心脏组织定居、生存 ,并与宿主心肌纤维排列方向一致 ,免疫组化检测胞质心肌特异蛋白染色阳性 ,与MI组比较 ,异体细胞移植组左室收缩压升高 (P <0 0 5) ,舒张末压明显降低 (P <0 0 1)、左心室内压最大上升和下降速率显著增快 (P <0 0 5) ,梗死边缘区心肌面毛细血管数目明显增加 (P <0 0 5) ,多功能真彩色病理图像分析系统显示MI面积缩小 (P <0 0 5)。结论　同种异体MSCs移植治疗MI可行、有效     

自体骨髓间充质干细胞移植对急性心肌梗死后心功能的影响

目的观察骨髓间充质干细胞（MSCs）经冠脉移植对急性心肌梗死后心功能的影响。方法24只日本大耳白兔,随机分为MSCs移植组（n=12）和培养液对照组（n=12）。从兔股骨抽取骨髓,体外培养MSCs。通过结扎左冠前降支建立急性心肌梗死模型。冠脉结扎后7d,细胞移植组和对照组直接经冠脉注入MSCs和培养液。于心肌梗死前、细胞移植前、细胞移植后1、2和4周对兔进行超声心动图检查。移植后4周处死动物,进行BrdU和第Ⅷ因子相关抗原免疫组化检测。结果移植后2周,MSCs移植组在射血分数（LVEF）和左室收缩末直径（LVESD）方面与移植前和对照组相比有显著改善（P<0.05）;移植4周后,LVEF、LVESD和左室舒张末直径（LVEDD）在MSCs移植组与移植前及对照组相比均有显著改善（P<0.05）。免疫组化检测发现,MSCs移植组BrdU染色阳性,血管计数较对照组明显增多（P<0.01）。结论经冠脉移植的MSCs可在梗死区心肌内存活并逐渐分化成心肌样细胞,促进毛细血管生成,显著改善心功能。     

骨髓间充质干细胞移植治疗心肌梗死的新技术

骨髓间充质干细胞移植是治疗心肌梗死的新途径之一,其技术和方法不断发展。诸如建立更好的大型活体实验动物模型；改进对植入细胞的标记追踪技术；进一步研究细胞植入的适宜方式和时间等。骨髓间充质干细胞移植和基因疗法相结合可能得到叠加的治疗效果。     

骨髓间充质干细胞移植治疗心肌梗死的新技术

骨髓间充质干细胞移植是治疗心肌梗死的新途径之一,其技术和方法不断发展.诸如建立更好的大型活体实验动物模型;改进对植入细胞的标记追踪技术;进一步研究细胞植入的适宜方式和时间等.骨髓间充质干细胞移植和基因疗法相结合可能得到叠加的治疗效果.     

骨髓间充质干细胞移植治疗心肌梗死的研究进展

骨髓间充质干细胞是一种多能干细胞,在体外培养时,可以通过诱导使其分化为心肌细胞等。目前进行的动物实验和临床Ⅰ期实验表明骨髓间充质干细胞具有促进血管增生以及改善心肌梗死后心脏功能的作用,为心肌梗死的治疗提供了广阔前景。     

骨髓间充质干细胞移植治疗心肌梗死机制的研究进展

心肌梗死（myocardial infarction,MD是由于冠状动脉循环改变引起冠状血流和心肌需求之间不平衡而导致的心肌损害,是临床上一种严重的缺血性心脏病（ischemic heart isease,IHD）。梗死的心肌细胞逐渐被瘢痕组织取代,由于瘢痕组织缺乏弹性,难以满足心脏收舒功能的要求,     

骨髓间充质干细胞静脉移植对心肌梗死大鼠心功能的影响

目的探讨静脉途径移植骨髓间充质干细胞(MSCs)对急性心肌梗死(AMI)大鼠心功能的影响。方法雄性SD大鼠20只,分为干细胞移植组(10只)和对照组(10只)。结扎大鼠左冠状动脉建立心肌梗死模型。同种异体大鼠MSCs体外分离、纯化、扩增,4’,6-二脒-2-苯基吲哚(4’,6-diamidino-2-phenylindole,DAPI)标记,并通过尾静脉于心肌梗死后1周移植入AMI大鼠体内,对照组则注射等量培养液。4周后测定心功能,并行免疫组织化学检测。结果心肌梗死4周后,干细胞移植组心肌组织冷冻切片中可以观察到DAPI标记细胞存在,免疫组织化学显示阳性标记细胞a-Actin肌动蛋白检测阳性。干细胞移植组心功能较对照组有明显改善(P<0.05)。结论MSCs经静脉移植可以归巢至梗死心肌处,并能改善受损的心功能。     

骨髓间充质干细胞经静脉移植到心肌梗死大鼠体内后的分布

目的 探讨经静脉注射骨髓间充质干细胞（MSCs）于心肌梗死大鼠模型后在体内的分布及对心功能的影响。方法 心肌梗死后1周将标记的MSCs注射到大鼠舌下静脉内,在细胞移植后不同时间点取心、肝、脾、肺、肾脏器,进行组织学检查观察移植细胞的分布。通过心脏超声复查移植后3周、6周心功能改变情况。结果 MSCs 经静脉注射后各脏器的组织结构无明显异常改变。移植细胞在早期主要分布在正常心肌组织内,1周后主要分布在梗死及交界区内,正常心肌组织内很少见细胞存留。细胞移植后3周、6周实验组左心室没有进一步扩大,心功能没有进一步恶化。结论 静脉注射移植MSCs后细胞可以分布到重要组织器官内,并不影响其组织结构;移植细胞有向梗死心肌组织内迁移的趋势,能明显延缓左心室重构而导致的心功能恶化。     

大鼠骨髓间充质干细胞移植在心肌梗死区长期存活的观察

目的 探讨大鼠骨髓间充质干细胞(BMSC)异体移植在心肌梗死区后长期存活数量的动态变化及其分布情况.方法 分离、纯化、培养雄性SD大鼠BMSC.开胸结扎雌性SD大鼠左冠状动脉前降支,于心肌梗死1 b后,将雄性大鼠BMSC 1.2×106个注入移植在雌性大鼠梗死心肌中心区心外膜下.将细胞移植受体大鼠随机分为3组,分别于移植后1、3和6周采集心肌梗死组织标本(n=18).实验动物移植细胞用Hoechst33342(n=5)和Brdu预先标记(n=5).实时定量PCR(realtime quantitative PCR)检测大鼠Y染色体雄性鉴别基因sry片段的表达(n=8).结果 分离纯化的雄性大鼠BMSC经流式细胞术鉴定为CID44+/CD34-.Hoechst33342荧光及Brdu免疫组织化学定性定位了移植细胞的存活,并在3周和6周组观察到部分标记细胞有序聚集排列,形成血管样结构.RT-PCR检测移植后细胞存活率在1、3和6周组分别为7.88%、7.82%和8.73%.结论 BMSC可以在心肌梗死区长期存活并参与受损心肌的修复.存活数量于移植1周后不再进行性减少,至6周均为8%左右.

Abstract：

Objective To investigate the quantification and distribution of survival transplanted bone marrow mesenchymal stem cells (BMSCs) in infarcted myocardium of rats. Methods BMSCs of male SD rats were isolated, purified and proliferated. Female SD rats were subjected to occlusion of left anterior descending artery, 1.2 × 106 BMSCs were injected into the centre zone of the infarct region 1 h after left anterior descending artery occlusion. Myocardial tissue was harvested randomly at 1,3 and 6 ( n = 18) weeks after transplantation. Transplanted cells were labeled with Hoechst33342 ( n = 5 ) and Brdu ( n = 5 ). Sry suquence of Y chromatosome in male rat was analyzed by real-time PCR in the tissue ( n = 8). Results FACS analysis showed that BMSCs expressed homogenous with CD44 +/ CD34 -. Hoechst33342 or Brdu labeled positive cells were found in localized accumulation and some of them migrated to the margin of the infarct region, and vessel-like structure could be observed in the host hearts. The real-time PCR demonstrated that the survival rates of engrafted BMSCs were 7. 88%, 7. 82% and 8. 73% respectively at 1,3 and 6 weeks after transplantation. Conclusion BMSCs could survive in the host infarcted myocardium for more than one month and the survival rate was around 8% from the first week to the sixth week after transplantation.     

EDG1 receptor stimulation leads to cardiac hypertrophy in rat neonatal myocytes

Sphingosine 1 phosphate (S1P), an aminophospholipid, acts extracellularly as a ligand via the specific G protein-coupled receptors of the endothelial differentiation gene (EDG) 1, 3, 5, 6 and 8 receptors family and intracellularly as a second messenger in various cellular types. The aim of this work was to investigate biological activity of S1P in cardiomyocytes with respect to related sphingolipids. S1P was applied for 48 h on rat neonatal cardiomyocytes at 10 nM, 100 nM and 1 microM. S1P induced a concentration-dependent cellular hypertrophy evidenced by an increase in cell size, [3H]-phenylalanine incorporation, protein content and Brain Natriuretic Peptide (BNP) secretion. Among the lipids tested S1P exhibits the lower EC50 (67 nM) followed by dihydro-S1P (107 nM) and sphingosylphosphorylcholine (1.6 microM). The effect of S1P could be related to a stimulation of the EDG1 receptor since we showed that the EDG1 receptor is predominantly expressed at the mRNA and protein levels in rat cardiomyocytes and that specific anti-EDG1 antibodies inhibited the hypertrophic effect induced by S1P. Furthermore the expression level of most other EDG receptors for S1P appeared very low in cardiac myocytes. S1P (100 nM) increased the phosphorylation of p42/44MAPK, p38MAPK, JNK, Akt and p70(S6K), this effect being reversed by inhibitors of their respective phosphorylation which also rescue the hypertrophic phenotype. Finally, S1P stimulated actin stress fibre formation reverted by the Rho inhibitor, the C3 exoenzyme. Altogether, our results show that S1P induces cardiomyocyte hypertrophy mainly via the EDG1 receptor and subsequently via Gi through ERKs, p38 MAPK, JNK, PI3K and via Rho pathway.     

Transplantation of human fetal blood stem cells in the osteogenesis imperfecta mouse leads to improvement in multiscale tissue properties

Osteogenesis imperfecta (OI or brittle bone disease) is a disorder of connective tissues caused by mutations in the collagen genes. We previously showed that intrauterine transplantation of human blood fetal stem/stromal cells in OI mice (oim) resulted in a significant reduction of bone fracture. This work examines the cellular mechanisms and mechanical bone modifications underlying these therapeutic effects, particularly examining the direct effects of donor collagen expression on bone material properties. In this study, we found an 84% reduction in femoral fractures in transplanted oim mice. Fetal blood stem/stromal cells engrafted in bones, differentiated into mature osteoblasts, expressed osteocalcin, and produced COL1a2 protein, which is absent in oim mice. The presence of normal collagen decreased hydroxyproline content in bones, altered the apatite crystal structure, increased the bone matrix stiffness, and reduced bone brittleness. In conclusion, expression of normal collagen from mature osteoblast of donor origin significantly decreased bone brittleness by improving the mechanical integrity of the bone at the molecular, tissue, and whole bone levels.     

Transplantation of adipose derived stromal cells is associated with functional improvement in a rat model of chronic myocardial infarction

AIMS: To determine the effect of transplantation of undifferentiated and cardiac pre-differentiated adipose stem cells compared with bone marrow mononuclear cells (BM-MNC) in a chronic model of myocardial infarction. METHODS: Ninety-five Sprague-Dawley rats underwent left coronary artery ligation and after 1 month received by direct intramyocardial injection either adipose derived stem cells (ADSC), cardiomyogenic cells (AD-CMG) or BM-MNC from enhanced-Green Fluorescent Protein (eGFP) mice. The control group was treated with culture medium. Heart function was assessed by echocardiography and 18F-FDG microPET. Cell engraftment, differentiation, angiogenesis and fibrosis in the scar tissue were also evaluated by (immuno)histochemistry and immunofluorescence. RESULTS: One month after cell transplantation, ADSC induced a significant improvement in heart function (LVEF 46.3+/-9.6% versus 27.7+/-8% pre-transplant) and tissue viability (64.78+/-7.2% versus 55.89+/-6.3% pre-transplant). An increase in the degree of angiogenesis and a decrease in fibrosis were also detected. Although transplantation of AD-CMG or BM-MNC also had a positive, albeit smaller, effect on angiogenesis and fibrosis in the infarcted hearts, this benefit did not translate into a significant improvement in heart function or tissue viability. CONCLUSION: These results indicate that transplantation of adipose derived cells in chronic infarct provides a superior benefit to cardiac pre-differentiated ADSC and BM-MNC.     

Hepatitis C eradication with sofosbuvir leads to significant metabolic changes

AIM To assess the effect of sofosbuvir(SOF) based regimens on glycemic and lipid control.METHODS This is a retrospective analysis of hepatitis C virus(HCV)-infected patients treated and cured with a SOF regimen [SOF/ribavirin/interferon, SOF/simeprevir, or SOF/ledipasvir(LDV) ± ribavirin] from January 2014 to March 2015. Patients with hemoglobin A1C(HbA1C) and lipid panels within six months before and six months after therapy were identified and included in our study. Due to the known hemolytic effect of ribavirin, HbA1C was obtained a minimum of three months post-treatment for the patients treated with a ribavirin regimen. Medical history, demographics, HCV genotype, pre-therapy RNA, and liver biopsies were included in our analysis. The patients who started a new medication or had an adjustment of baseline medical management for hyper-lipidemia or diabetes mellitus(DM) were excluded from our analysis.RESULTS Two hundred and thirty-four patients were reviewed, of which 60 patients met inclusion criteria. Sixty-three point three percent were male, 26.7% were Caucasian, 41.7% were African American and 91.7% were infected with hepatitis C genotype 1. Mean age was 60.6 ± 6.7 years. Thirty-nine patients had HbA1C checked before and after treatment, of which 22 had the diagnosis of DM type 2. HbA1C significantly decreased with treatment of HCV(pretreatment 6.66% ± 0.95% vs posttreatment 6.14% ± 0.65%, P 0.005). Those treated with SOF/LDV had a lower HbA1C response than those treated with other regimens(0.26% ± 0.53% vs 0.71% ± 0.83%, P = 0.070). Fifty-two patients had pre- and post-treatment lipid panels; there was a significant increase in low-density lipoprotein(LDL) and total cholesterol(TC) after treatment(LDL: 99.5 ± 28.9 mg/dL vs 128.3 ± 34.9 mg/dL, P 0.001; TC: 171.6 ± 32.5 mg/dL vs 199.7 ± 40.0 mg/dL, P 0.001). Pretreatment body-mass index(BMI) did not differ from post-treatment BMI(P = 0.684). CONCLUSION Eradication of HCV with a SOF regimen resulted in a significant drop in HbA1C and an increase in LDL and TC post therapy.     

Transplantation of adipose tissue-derived stem cells improves cardiac contractile function and electrical stability in a rat myocardial infarction model

The transplantation of adipose tissue-derived stem cells (ADSCs) improves cardiac contractility after myocardial infarction (MI); however, little is known about the electrophysiological consequences of transplantation. The purpose of this study was to clarify whether the transplantation of ADSCs increases or decreases the incidence of ventricular tachyarrhythmias (VT) in a rat model of MI. MI was induced experimentally by permanent occlusion of the left anterior descending artery of Lewis rats. ADSCs were harvested from GFP-transgenic rats, and were cultured until passage four. ADSCs (10 × 10⁶) resuspended in 100 μL saline or pro-survival cocktail (PSC), which enhances cardiac graft survival, were injected directly into syngeneic rat hearts 1 week after MI. The recipients of ADSCs suspended in PSC had a larger graft area compared with those receiving ASDCs suspended in saline at 1 week post-transplantation (number of graft cells/section: 148.7 ± 10.6 vs. 22.4 ± 3.4, p < 0.05, n = 5/group). Thereafter, all ADSC recipients were transplanted with ASDCs in PSC. ADSCs were transplanted into infarcted hearts, and the mechanical and electrophysiological functions were assessed. Echocardiography revealed that ADSC recipients had improved contractile function compared with those receiving PSC vehicle (fractional shortening: 21.1 ± 0.9 vs. 14.1 ± 1.2, p < 0.05, n ≥ 12/group). Four weeks post-transplantation, VT was induced via in vivo programmed electrical stimulation. The recipients of ADSCs showed a significantly lower incidence of induced VT compared with the control (31.3% vs. 83.3%, p < 0.05, n ≥ 12/group). To understand the electrical activity following transplantation, we performed ex vivo optical mapping using a voltage sensitive dye, and found that ADSC transplantation decreased conduction velocity and its dispersion in the peri-infarct area. These results suggest that ADSC transplantation improved cardiac mechanical and electrophysiological functions in subacute MI.     

心脏植入式电子装置电极对心脏的血管损伤

心脏植入式电子装置(CIED)植入是一项安全有效的临床技术。在植入相关的并发症中,电极损伤心脏血管的发生率较低,但发生后可引起夹层、急性心肌梗死、心脏压塞、动脉瘤及动脉瘘等严重并发症及临床后果,故应引起临床医师的高度重视。本文对CIED植入术中及术后起搏器和除颤电极损伤冠状动脉、冠状静脉及升主动脉的相关报道进行综述。