CD90 心脏成纤维细胞减少大鼠急性心肌损伤的纤维化。

CD90 cardiac fibroblasts reduce fibrosis of acute myocardial injury in rats.

机构信息

Henan Key Laboratory of Medical Tissue Regeneration, Xinxiang Medical University, Xinxiang 453003, PR China.

出版信息

Int J Biochem Cell Biol. 2018 Mar;96:20-28. doi: 10.1016/j.biocel.2018.01.009. Epub 2018 Jan 12.

DOI:10.1016/j.biocel.2018.01.009

PMID:29339198

Abstract

OBJECTIVE

To explore the differentiation tendency of CD90 cardiac fibroblast (CFs) into cardiomyogenic cells in vitro and repair functions in acute myocardial infarction rats.

METHODS

CD90 subpopulation was sorted from rat CFs by flow cytometry. 10 μmoL/L of 5-Azacytosine (5-aza) was used to induce differentiation of CFs into cardiomyogenic cells. An acute myocardial infarction model was prepared by ligation of the rat left anterior descending coronary artery. After nuclei were labeled by DAPI, induced CD90 CFs were injected into the infarction marginal zone. Before coronary ligation, 40 min after ligation, and at 7 and 14 days after cell transplantation, cardiac function changes were detected by ultrasound imaging system respectively. cTnT and endothelial cell marker VIII factor were detected by immunofluorescence staining. Infarct size was examined by TTC staining. Fibrosis was evaluated with masson's trichrome staining, vimentin, type I and type III collagen staining.

RESULTS

CD90 CFs sorted by flow cytometry was 34.9%. On day 28 after induction, the cTnT positive rate was 61.17 ± 9.75%. Left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) at days 7 and 14 after transplantation were significantly increased compared with those before transplantation (P < 0.05). LVEF and LVFS at the 14th day after transplantation were also significantly increased compared with those at the 7th day (LVEF: 61.40 ± 2.45% vs. 56.25 ± 2.9%, LVFS: 33.21 ± 0.68% vs. 30.26 ± 2.06%, P < 0.01). Additionally, small numbers of CD90 CFs differentiated into cardiomyocytes and became involved in neovascularization. CD90 CFs and CFs reduced myocardial infarct size at days 14. It was significantly smaller in rats with CFs transplantation group than those in MI group(24.78 ± 2.28% vs. 31.28 ± 2.83%, P < 0.05), and it was also significantly smaller in rats with CD90CFs transplantation group than those in CFs transplantation group (17.47 ± 4.15% vs. 24.78 ± 2.28%, P < 0.05). Meanwhile, The percentage of fibrotic area and vimentin, type I and type III collagen in the infarct border zone and infarct area were both significantly reduced in CD90 + CFs.

CONCLUSION

CD90 CFs is the preponderant subpopulation of cardiomyogenic differentiation, with potential use as seed cells in basic and clinical research of heart regeneration and repair.

摘要

目的

探讨 CD90 心脏成纤维细胞（CFs）在体外向心肌样细胞分化的趋势及其在急性心肌梗死大鼠中的修复功能。

方法

采用流式细胞术分选 CD90 亚群。用 10μmol/L 的 5-氮杂胞苷（5-aza）诱导 CFs 向心肌样细胞分化。结扎大鼠左前降支冠状动脉制备急性心肌梗死模型。用 DAPI 标记细胞核后，将诱导的 CD90 CFs 注入梗死边缘区。在冠状动脉结扎前、结扎后 40min 及细胞移植后 7、14d 分别用超声成像系统检测心功能变化。用免疫荧光染色检测 cTnT 和内皮细胞标记物 VIII 因子。用 TTC 染色检测梗死面积。用 Masson 三色染色、波形蛋白、I 型和 III 型胶原染色评估纤维化。

结果

用流式细胞术分选的 CD90 CFs 占 34.9%。诱导 28d 后，cTnT 阳性率为 61.17±9.75%。移植后 7、14d 的左心室射血分数（LVEF）和左心室短轴缩短率（LVFS）均明显高于移植前（P<0.05）。移植后 14d 的 LVEF 和 LVFS 也明显高于 7d（LVEF：61.40±2.45%比 56.25±2.9%，LVFS：33.21±0.68%比 30.26±2.06%，P<0.01）。此外，少量 CD90 CFs 分化为心肌细胞并参与新生血管形成。CD90 CFs 和 CFs 在第 14 天减少了心肌梗死面积。CFs 移植组大鼠的梗死面积明显小于 MI 组（24.78±2.28%比 31.28±2.83%，P<0.05），CD90 CFs 移植组大鼠的梗死面积也明显小于 CFs 移植组（17.47±4.15%比 24.78±2.28%，P<0.05）。同时，梗死边缘区和梗死区的纤维化面积百分比和波形蛋白、I 型和 III 型胶原均明显减少。