分泌IGF-1的间充质干细胞对肝脏缺血-再灌注损伤的保护作用

The protective effect of IGF-1-secreting mesenchymal stromal cell on liver ischemia-reperfusion injury

目的探讨分泌胰岛素样生长因子1的间充质干细胞(IGF-1-MSC)对小鼠肝脏缺血-再灌注损伤(IRI)的作用。方法 50只C57BL/6系小鼠按随机数字表法随机分为IRI组、PBS组、MSC组、IGF-1-MSC组、对照组,各10只。建立小鼠肝脏IRI模型,缺血时间60 min,再灌注时IRI组不做处理,PBS组经门静脉注射PBS溶液100μl,MSC组经门静脉注射MSC细胞悬液100μl,IGF-1-MSC组经门静脉注射IGF-1-MSC细胞悬液100μl。检测AST、ALT、乳酸脱氢酶(LDH)水平,并行肝组织病理学检查及TUNEL染色。各组检测指标比较采用单因素分析和LSD-t检验或Kruskal-Wallis检验。结果再灌注后24 h,IGF-1-MSC组AST、ALT、LDH分别为(815±233)、(867±350)、(845±358)U/L,明显低于MSC组的(3 805±1 355)、(2 421±845)、(2 011±162)U/L(LSD-t=-4.865,-3.725,-6.637;P<0.05)。IGF-1-MSC组肝窦轻度淤血,少量肝细胞凋亡;而IRI组肝细胞大量凝固性坏死和细胞凋亡。IGF-1-MSC组24 h肝组织损伤Suzuki评分为4(3~4)分,明显低于MSC组和IRI组的5(4~5)和8(7~10)分(Z=-2.545,-2.703,P<0.05)。结论 IGF-1-MSC输注能有效减轻肝脏IRI程度,减少肝细胞凋亡可能是其保护作用机制之一。

Objective To investigate the effect of insulin-like growth tactor l-secreung mesenchymal stromal cell （IGF-1-MSC） on liver ischemia-reperfusion injury （IRI） in mice. Methods According to the random number table method, 50 C57BL/6 mice were randomly divided into the IRI group, PBS group, MSC group, IGF-1-MSC group and control group with 10 mice itt each group. The hepatic IRI models of mice were established and the ischemia time was 60 min. During reperfusion, no interventions were delivered in the IRI group, the mice were injected with 100 μl PBS through portal vein in the PBS group, 100 μl MSC suspension in the MSC group and 100 μl IGF-1-MSC suspension in the IGF-1-MSC group. The levels ofAST, ALT and lactic dehydrogenase （LDH） were detected. Liver tissues were collected for pathological examination and TUNEL staining. The indexes of the groups were compared using one-way analysis of variance and LSD-t test or Kruskal-Wallis test. Results The level of AST, ALT and LDH at 24 h after reperfusion in the IGF-1-MSC group was respectively （815±233）, （867±350） and （845±358） U/L, significantly lower than （3 805±1 355）, （2 421±845） and （2 011±162） U/L in the MSC group （LSD-t=--4.865, -3.725, -6.637; P〈0.05）. Mild sinusoid congestion and a small amount of hepatocyte apoptosis were observed in the IGF-1-MSC group, while a large number of hepatocyte coagnlative necrosis and apoptosis were observed in the IRI group. The liver tissue injury Suzuki score at 24 h in the IGF-1-MSC group was 4（3-4）, significantly lower than 5（4-5） in the MSC group and 8（7-10） in the IRI group （Z=-2.545, -2.703; P〈0.05）. Conclusions Injection of IGF-1-MSC can effectively alleviate the severity of liver IRI. Reducing hepatocyte cell apoptosis may probably be one of the mechanisms of the protective role of IGF-1-MSC.