妊娠大鼠来源脂肪干细胞在急性肝损伤中的修复作用

发布时间：2019-01-30 18:26

【摘要】：目的评估妊娠大鼠来源脂肪干细胞(adipose-derived stem cells,ADSCs)在急性肝损伤中的修复作用。方法取18周龄妊娠SD大鼠分离培养ADSCs并鉴定。另取SD大鼠20只随机分为A、B、C、D 4组(n=5),A组大鼠不作处理,作为正常对照;B~D组大鼠腹腔注射CCl4制备急性肝损伤模型。造模后2 h,A组于大鼠脾脏注射DPBS,B组不注射任何试剂,C、D组分别于大鼠脾脏注射0.1 mL正常大鼠来源和妊娠大鼠来源ADSCs(2×10~6个/mL)。注射后7 d处死动物,取眼球血检测血清中总胆红素(total bilirubin,TBIL)、丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartic acid transaminase,AST)、白蛋白(albumin,ALB)和肝功能总蛋白(total protein,TP)含量;取肝组织切片行HE染色检测损伤肝组织形态学变化;免疫组织化学染色检测肝组织中Ki67、甲胎蛋白(alpha-fetoprotein,AFP)和ALB的表达情况。结果 B组血清中TBIL、ALT、AST含量显著高于A、C、D组,ALB、TP含量显著低于A、C、D组,差异均有统计学意义(P0.05)。C、D组血清中TBIL、ALT、AST含量显著高于A组,C组高于D组,差异均有统计学意义(P0.05);A、C、D组血清中ALB含量差异无统计学意义(P0.05);C、D组血清中TP含量显著低于A组(P0.05),C、D组间比较差异无统计学意义(P0.05)。HE染色示:A组大鼠肝组织结构清晰,细胞排列整齐,大小均匀;B组大鼠肝细胞明显出现水肿,细胞排列紊乱,可见肝小叶内点状坏死,组织出现弥漫性炎性细胞浸润;C、D组大鼠肝组织炎症和肝细胞坏死程度较B组明显减轻,同时由线粒体和粗面内质网扩张造成的空泡数量降低,其中D组肝损伤改善程度较C组更显著。免疫组织化学染色示:C、D组Ki67阳性细胞率显著高于A、B组,B组高于A组,D组高于C组,差异均有统计学意义(P0.05)。A、B组AFP均无表达,而C、D组阳性表达,D组AFP阳性细胞率显著高于C组(t=3.006,P=0.017)。C、D组ALB表达均显著高于A、B组,D组高于C组,差异均有统计学意义(P0.05)。结论与正常大鼠来源ADSCs移植比较,妊娠大鼠来源ADSCs移植后,肝损伤大鼠肝功能改善程度更显著。
[Abstract]:Objective to evaluate the repair effect of adipose stem cells (adipose-derived stem cells,ADSCs) from pregnant rats in acute liver injury. Methods ADSCs was isolated from 18 week pregnant SD rats and identified. In addition, 20 SD rats were randomly divided into two groups: group A (n = 5), A) and group B (n = 5), A), and group B (n = 20) were injected intraperitoneally with CCl4 to make acute liver injury model. 2 hours after modeling, group A did not inject any reagents into the spleen of rats injected with DPBS,B. Group C D injected 0.1 mL ADSCs from normal rats and pregnant rats from ADSCs (2 脳 106 / mL).), respectively. Seven days after injection, the animals were killed and their eyeball blood was taken to detect serum total bilirubin (total bilirubin,TBIL), alanine aminotransferase (alanine aminotransferase,ALT), aspartate aminotransferase (aspartic acid transaminase,AST), albumin (albumin,). ALB and total liver protein (total protein,TP) content; The morphological changes of injured liver tissue were detected by HE staining and the expression of Ki67, alpha fetoprotein (alpha-fetoprotein,AFP) and ALB in liver tissue were detected by immunohistochemical staining. Results the levels of serum TBIL,ALT,AST and ALB,TP in group B were significantly higher than those in group A (P 0.05), and the levels of ALB,TP in group C were significantly lower than those in group D (P0.05). The content of TBIL,ALT,AST in group C was significantly higher than that in group A (P 0.05). Group C was higher than group D, the difference was statistically significant (P0.05). There was no significant difference in serum ALB content in group A (P 0.05). The content of serum TP in group C D was significantly lower than that in group A (P0.05), but there was no significant difference between group C and D (P0.05). HE staining showed that the liver tissue structure of group A was clear, cells were arranged neatly, and the size was uniform; In group B, there were edema and disarrangement of hepatocytes, dotted necrosis and diffuse inflammatory cell infiltration in liver lobule. The degree of liver inflammation and hepatocyte necrosis in group C was significantly reduced than that in group B, and the number of vacuoles caused by dilatation of mitochondria and rough endoplasmic reticulum was decreased. The improvement of liver injury in group D was more significant than that in group C. Immunohistochemical staining showed that the rate of Ki67 positive cells in group C was significantly higher than that in group A, group B and group C, respectively (P 0.05). There was no expression of AFP in group C, but no expression of AFP in group C was found in group C (P0.05), but the expression of AFP in group C was significantly higher than that in group C (P0.05). The positive expression rate of AFP in group D was significantly higher than that in group C (t 3.006). The expression of ALB in group C was significantly higher than that in group A (P 0.05). The expression of ALB in group D was significantly higher than that in group C (P0.05). Conclusion compared with the normal rat ADSCs transplantation, the liver function of the gravid rats was improved more significantly after the transplantation of the ADSCs derived from the pregnant rats.
【作者单位】：昆明医科大学第四附属医院骨与创伤外科;昆明医科大学第二附属医院急诊医学部;
【分类号】：R575

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