大黄游离蒽醌对脂肪干细胞移植治疗大鼠急性胰腺炎的影响  

Effect of rhubarb-free anthraquinone in the treatment of acute pancreatitis with adipose-derived stem cell transplantation in rats

作　　者：郑俊杰[1] 田伟军[2] 王金峰 叶奎[4] Zheng Junjie;Tian Weijun;Wang Jinfeng;Ye Kui(Department of hepatobiliary and gastrointestinal surgery,Tianjin fourth central hospital,Tianjin 300140;Department of General Surgery,General Hospital of Tianjin Medical University,Tianjin 300052,China;Second Department of Surgery,Liaoning Electric Power Center Hospital,Shenyang 110000,Liaoning Province,China;Department of vascular surgery,Tianjin fourth central hospital,Tianjin 300140)

机构地区：[1]天津市第四中心医院肝胆胃肠外科,天津市300140 [2]天津医科大学总医院普外科,天津市300052 [3]辽宁电力中心医院外二科,辽宁省沈阳市110000 [4]天津市第四中心医院血管外科,天津市300140

出　　处：《中国组织工程研究》2019年第9期1390-1396,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:研究表明大黄游离蒽醌能够保护肠黏膜细胞结构和功能的稳定,但尚未有将其与脂肪干细胞联合治疗大鼠急性胰腺炎的报道。目的:观察并探讨大黄游离蒽对脂肪干细胞移植治疗大鼠急性胰腺炎的影响及其机制。方法:体外快速复苏并培养购自中国医学科学院的冻存鼠脂肪干细胞,随后采用MTT比色法测定脂肪干细胞的细胞存活率并行PKH-26标记。将北京维通利华动物实验技术有限公司提供的SD大鼠随机平均分为4组,病理模型组大鼠采用肠壁穿刺逆行胰胆管注射体积分数5%牛黄胆酸钠建立急性胰腺炎病理模型,然后尾静脉注射0.5 mL的L-DMEM完全培养液;大黄游离蒽组建模后给予大黄游离蒽醌200 mg/kg灌胃;脂肪干细胞组建模后尾静脉注射0.5 mL细胞浓度1×10~7 L^(-1)脂肪干细胞;联合治疗组建模后同时给予以上2种干预。各个实验组中所有干预治疗措施每日进行1次且连续治疗3 d。结果与结论:(1)与理模型组的大鼠相比,大黄游离蒽醌组和脂肪干细胞组大鼠血清淀粉酶活性以及白细胞介素6水平显著降低(P <0.05),胰蛋白酶原激活肽水平显著升高(P <0.05);经过脂肪干细胞移植和大黄游离蒽醌灌胃处理联合治疗,上述3指标水平较大黄游离蒽醌组和脂肪干细胞组进一步降低(P<0.05);(2)苏木精-伊红染色结果显示,此2个单独治疗组大鼠胰腺组织中脂肪变性、出血、细胞坏死及炎症细胞浸润等病理变化程度均有明显减轻,联合治疗组中大鼠的病理变化的缓解程度则更为明显;(3)荧光显微镜下观察可见,PKH-26标记的阳性细胞数联合治疗组最多,脂肪干细胞组次之,而在大黄游离蒽组及病理模型组则未见(P <0.05);(4)TUNEL法检测可见,与病理模型组比较,2个单独治疗组中胰腺组织中的细胞凋亡数明显降低,联合治疗组进一步降低(P <0.01);(5)RT-PCR和Western blot法检测显示,病理模型组大鼠胰腺组织转�BACKGROUND:Studies have shown that rhubarb-free anthraquinone can protect the stability of the intestinal mucosa cell structure and function.However,the combination of rhubarb-free anthraquinone with adipose-derived stem cells to treat acute pancreatitis in rats has not yet been reported.OBJECTIVE:To investigate the effects of rhubarb-free anthraquinone on the treatment of acute pancreatitis by fat stem cell transplantation in rats.METHODS:Cryopreserved mouse adipose-derived stem cells(provided by the Chinese Academy of Medical Sciences)were rapidly recovered and cultured in vitro.The survival cells were labeled using PKH-26,and then the cell survival rate of adipose-derived stem cells was determined by MTT colorimetry.Sprague-Dawley rats provided by Beijing Vital River Laboratory Animal Technology Co.,Ltd.were equally randomized into four groups.Animal models of acute pancreatitis were made in all rats through intestinal wall puncture for retrograde administration of 5%sodium taurocholate to the pancreaticobiliary tube,and then the model rats were given tail vein injection of L-DMEM complete medium(0.5 mL)in the model group,intragastric administration of 200 mg/kg rhubarb-free anthraquinone in the rhubarb-free anthraquinone group,tail vein injection of adipose-derived stem cells(1×107 cells/L,0.5 mL)in the cell transplantation group,and intragastric administration of 200 mg/kg rhubarb-free anthraquinone plus tail vein injection of 1×107 adipose-derived stem cells(0.5 mL)in the combined treatment group.All interventions in each group were performed once a day,for 3 continuous days.RESULTS AND CONCLUSION:(1)The levels of serum amylase and interleukin-6 were significantly reduced in the rhubarb-free anthraquinone group and cell transplantation group compared with the model group(P<0.05),while the level of trypsinogen activation peptide significantly increased(P<0.05).In the combined treatment group,the levels of three indicators mentioned above were significantly decreased compared with the rhubarb-free anthraqui

关 键 词：脂肪干细胞 移植 大鼠 急性胰腺炎 大黄游离蒽 转化生长因子β/Smad 胰腺组织 血清淀粉酶 白细胞介素6 胰蛋白酶原激活肽 凋亡 胰腺炎 干细胞 组织工程 

分 类 号：R456[医药卫生—治疗学] R3648[医药卫生—临床医学]