骨髓间充质干细胞对重症急性胰腺炎胰腺组织修复的促进作用研究

目的观察骨髓间充质干细胞（BMSCs）抑制坏死性凋亡促进修复重症急性胰腺炎（SAP）的可能机理。 方法（1）分离、培养和鉴定大鼠BMSCs;（2）构建牛磺胆酸钠（NaT）诱导的SAP大鼠模型,并分成正常组（NC）、假手术组（Sham）、SAP模型组（SAP）、PBS治疗组（PBS）、BMSCs治疗组和Necrostain-1 （Nec-1）治疗组,并检测胰腺病理评分和血淀粉酶水平;（3）运用Western Blot和qRT-PCR方法检测各组受损胰腺组织内RIPK1、RIPK3、Caspase-8、MLKL蛋白及mRNA表达,各组均数间比较采用单因素方差分析,两两比较采用LSD-t检验。 结果BMSCs可以被诱导分化成骨、软骨、脂肪,并高表达CD44 （99.82﹪）、CD73 （99.87﹪）、CD90 （99.99﹪）、CD105 （99.78﹪）,低表达CD11b （0.65﹪）、CD19 （0.85﹪）、CD34 （0.70﹪）和CD45 （1.20﹪）。SAP组胰腺病理评分（12.90±1.79）及血淀粉酶水平（1052.41±183.12） mU/ ml均高于NC组评分：0.40±0.52,淀粉酶水平：（236.62±33.21） mU/ ml]和Sham组评分：0.50±0.53,淀粉酶水平：（242.31±27.94） mU/ ml]（F = 200.275,F = 143.245,P均< 0.001）,且SAP组受损胰腺组织内RIPK1、RIPK3、MLKL表达升高、Caspase-8表达降低（F = 179.905,P < 0.001）;BMSCs组和Nec-1治疗组胰腺病理评分及血淀粉酶水平均低于PBS治疗组评分：7.20±1.23、7.00±1.05比12.60±1.65,F = 200.275,P < 0.001;淀粉酶水平：（452.21±101.68）mU/ml、（570.18±148.47） mU/ml比（972.77±204.29） mU/ml,F = 143.245,P < 0.001],同时受损胰腺组织内RIPK1、RIPK3、MLKL表达下调、Caspase-8表达升高（F = 179.905,P < 0.001）。 结论BMSCs可能通过抑制坏死性凋亡通路活化来修复SAP。

Positive effect of bone marrow-derived mesenchymal stem cells on the repair of pancreatic tissue in severe acute pancreatitis

ObjectiveTo investigate the possible mechanism of bone marrow mesenchymal stem cells (BMSCs) promoting the repair of severe acute pancreatitis (SAP) by inhibiting necropotosis. Methods(1) the isolation, culture and identification of rat BMSCs; (2) the rat SAP models induced by sodium sulfonycholate (NaT) were constructed and divided into the normal group (NC) , Sham group (Sham) , SAP model group, PBS treatment group, BMSCs transplantation group and the Necrostain-1 (Nec-1) treatment group. (3) The Western-blot and qRT-PCR were used to detect the protein and mRNA expressions of RIPK1, RIPK3, caspase-8 and MLKL in the damaged pancreatic tissues of each group. One-way anova was used for comparison between the mean values of each group, and LSD-t test was used for pair comparison. ResultsBMSCs could be induced to differentiate into bone, cartilage and fat, with high expressions of CD44 (99.82﹪) , CD73 (99.87﹪) , CD90 (99.99﹪) and CD105 (99.78﹪) , low expressions of CD11b (0.65﹪) , CD19 (0.85﹪) , CD34 (0.70﹪) and CD45 (1.20﹪) . Pancreatic pathology scores (12.90±1.79) and blood amylase level (1052.41±183.12) mU/ml in SAP model group were significantly higher than those in NC group scores: 0.40±0.52, amylase level: (236.62±33.21) mU/ml] and Sham group scores: 0.50±0.53, amylase level: (242.31±27.94) mU/ml] (F = 200.275, F = 143.245, P < 0.001) . Moreover, the expressions of RIPK1, RIPK3 and MLKL were significantly increased and the expression of caspase-8 was significantly decreased in the SAP model group (F = 179.905, P < 0.001) . Pancreatic pathology scores and blood amylase level in BMSCs transplantation group and Nec-1 treatment group were significantly lower than those in PBS treatment group score: 7.20±1.23, 7.00±1.05 vs 12.60±1.65, F = 200.275, P < 0.001; Amylase levels: (452.21±101.68) mU/ ml, (570.18±148.47) mU/ml vs (972.77±204.29) mU/ml, F = 143.245, P < 0.001], while the expressions of RIPK1, RIPK3, MLKL were significantly down-regulated and the expression of caspase-8 was significantly increased in the damaged pancreas (F = 179.905, P < 0.001) . ConclusionBMSCs may repair SAP by inhibiting the activation of Necroptosispathway .