Tang W P, Hu Q, Chen G D, Luo J X
Division of Hepatobiliary and Pancreatic Surgery, First Affiliated Hospital of University of South China, Hengyang 421001, China.
Zhonghua Gan Zang Bing Za Zhi. 2020 Dec 20;28(12):1030-1035. doi: 10.3760/cma.j.cn501113-20190407-00113.
To observe the effect of basic fibroblast growth factor (bFGF) treatment on efficacy of adipose-derived mesenchymal stem cells (ADSCs) in cirrhotic rats. A rat model of liver cirrhosis was established via intraperitoneal injection of carbon tetrachloride (CCl(4)) for 10 weeks. Thirty SD rats were randomly divided into 3 groups ( = 10): control group served as group A, and 0.5ml of phosphate-buffered saline (PBS) was transfused into the tail vein; ADSCs single-dose transplantation group served as group B, and 1×10(6) ADSCs were transplanted into the tail vein; bFGF-treated ADSCs treatment group served as group C, and 1×10(6) bFGF-treated ADSCs were transplanted through tail vein. Liver function, pathological and cytokine changes, and the in vivo survival transformation condition of the transplanted cells were measured at one week after transplantation. F test and an independent sample t test were used. bFGF treatment had significantly promoted the proliferation, differentiation and overexpression of hepatocyte growth factor (HGF) in ADSCs [ADSCs single: (2 137.16 ± 261.52) pg/ml vs. ADSCs (bFGF): (4 776.23 ± 532.44) pg/ml, < 0.05]. The bFGF-treated ADSCs treatment group had statistically significant differences in promoting the recovery of liver function [alanine aminotransferase (ALT): ADSCs single (190.8 ± 34.98) vs. ADSCs (bFGF): (117.8 ± 35.81) pg/ml; aspartate aminotransferase (AST): ADSCs single (295.2 ± 33.71) U/L vs. ADSCs (bFGF): (183.8 ± 41.29) U/L, < 0.05). There was no statistically significant difference in serum albumin levels between the control group, ADSCs single group, and ADSCs (bFGF) group. Compared with the control group, the serum albumin level of ADSCs (bFGF) group was increased significantly ( < 0.05), and the difference between the control group and the ADSCs single transplantation group in improving liver regeneration and reducing liver damage was statistically significant ( < 0.05). Masson trichrome staining showed that the percentage of the liver fibrosis area in the bFGF-treated ADSCs treatment group was 6.78% ± 0.56%, which was significantly higher than that of the control and ADSCs single dose transplantation group (7.96% ± 0.64%) ( < 0.05). Western blot analysis showed that the expression of HGF protein in the bFGF-treated ADSCs treatment group was significantly up-regulated, while the expression of α-smooth muscle actin was significantly down-regulated, and the difference was significant in the ADSCs single transplantation group. A double fluorescent staining method showed that the numbers of stem cells implanted in the liver tissue of the bFGF-treated ADSCs treatment group were higher than that of the ADSCs single transplantation group. In-vitro cell experiments confirmed that bFGF had significantly promoted the overexpression of HGF in ADSCs. bFGF-treated ADSCs transplantation can significantly improve liver function and fibrosis as compared with ADSCs single-dose transplantation in cirrhotic rats.
观察碱性成纤维细胞生长因子（bFGF）处理对脂肪间充质干细胞（ADSCs）治疗肝硬化大鼠疗效的影响。通过腹腔注射四氯化碳（CCl₄）10周建立肝硬化大鼠模型。将30只SD大鼠随机分为3组（每组n = 10）：对照组为A组，尾静脉输注0.5ml磷酸盐缓冲液（PBS）；ADSCs单剂量移植组为B组，尾静脉移植1×10⁶个ADSCs；bFGF处理的ADSCs治疗组为C组，尾静脉移植1×10⁶个经bFGF处理的ADSCs。移植后1周检测肝功能、病理及细胞因子变化，以及移植细胞的体内存活转化情况。采用F检验和独立样本t检验。bFGF处理显著促进了ADSCs中肝细胞生长因子（HGF）的增殖、分化和过表达[ADSCs单剂量组：（2 137.16 ± 261.52）pg/ml vs. ADSCs（bFGF）组：（4 776.23 ± 532.44）pg/ml，P < 0.05]。bFGF处理的ADSCs治疗组在促进肝功能恢复方面有统计学显著差异[丙氨酸氨基转移酶（ALT）：ADSCs单剂量组（190.8 ± 34.98）vs. ADSCs（bFGF）组：（117.8 ± 35.81）pg/ml；天冬氨酸氨基转移酶（AST）：ADSCs单剂量组（295.2 ± 33.71）U/L vs. ADSCs（bFGF）组：（183.8 ± 41.29）U/L，P < 0.05]。对照组、ADSCs单剂量组和ADSCs（bFGF）组血清白蛋白水平无统计学显著差异。与对照组相比，ADSCs（bFGF）组血清白蛋白水平显著升高（P < 0.05），对照组与ADSCs单剂量移植组在改善肝再生和减轻肝损伤方面的差异有统计学意义（P < 0.05）。Masson三色染色显示，bFGF处理的ADSCs治疗组肝纤维化面积百分比为6.78% ± 0.56%，显著低于对照组和ADSCs单剂量移植组（7.96% ± 0.64%）（P < 0.05）。蛋白质免疫印迹分析显示，bFGF处理的ADSCs治疗组HGF蛋白表达显著上调，而α-平滑肌肌动蛋白表达显著下调，在ADSCs单剂量移植组差异显著。双荧光染色法显示，bFGF处理的ADSCs治疗组肝组织中植入的干细胞数量高于ADSCs单剂量移植组。体外细胞实验证实，bFGF显著促进了ADSCs中HGF的过表达。与ADSCs单剂量移植相比，bFGF处理的ADSCs移植可显著改善肝硬化大鼠的肝功能和纤维化。
