骨髓间充质干细胞治疗大鼠慢性胰腺炎的实验研究

目的探讨骨髓间充质干细胞(BMSCs)对大鼠慢性胰腺炎(CP)的治疗作用及其机制。方法将SD大鼠随机分为空白组(尾静脉注射无菌生理盐水),模型组(尾静脉注射二氯二丁基酯制作大鼠CP模型),治疗组(造模后尾静脉注射BMSCs),假治疗组(造模后尾静脉注射无菌生理盐水),每组10只。取大鼠胰腺组织进行病理学评分及纤维化程度评分,ELASA法检测胰腺组织中胰腺星状细胞(PSC)活化表达的α-平滑肌肌动蛋白(α-SMA),I型胶原蛋白,瓜型胶原蛋白水平,白细胞介素10(IL-10)水平;qRT-PCR法检测胰腺组织中PSC凋亡基因BNIP3的表达,Western-blot法检测胰腺组织中TGF-β/Smad信号传导通路中TGF-β1、Smad2、Smad3、Smad4的表达水平。将CP大鼠PSC培养体系分为对照组(单纯PSC培养组)和治疗组(PSC和BMSCs共培养组),每组5份标本,ELASA法检测α-SMA,I型胶原蛋白,Ⅲ型胶原蛋白,IL-10的表达水平;qRT-PCR法检测PSC凋亡基因BNIP3的表达,Western-blot法检测TGF-β1、Smad2、Smad3、Smad4的表达水平。结果(1)治疗组大鼠胰腺病理学评分及纤维化程度评分均低于模型组与假治疗组,且差别有统计学意义(P<0.05);治疗组α-SMA、I型胶原蛋白、Ⅱ型胶原蛋白水平,TGF-βl、Smad2、Smad3、Smad4的表达水平均低于模型组与假治疗组,且差别有统计学意义(P<0.05),但PSC凋亡基因BNIP3以及IL-10的表达水平高于模型组与假治疗组,且差别有统计学意义(P<0.05);(2)治疗组α-SMA、I型胶原蛋白、Ⅰ型胶原蛋白水平,TGF-β1、Smad2、Smad3、Smad4的表达水平均低于对照组,且差别有统计学意义(P<0.05),但PSC凋亡基因BNIP3以及IL-10的表达水平高于对照组,且差别有统计学意义(P<0.05)。结论BMSCs可通过TGF-β/Smad信号传导通路抑制PSC增殖活化,减少胰腺纤维化的形成,进而控制慢性胰腺炎。

The experimental study of bone marrow mesenchymal stem cells for the treatment of chronic pancreatitis in rats

Objective To investigate the effects and mechanisms of BMSCs on CP in rats.Methods 40 SD rats were divided into 4 groups with 10 in each:control group,model group,treatment group,and sham treatment group.The pancreatic fibrosis and pathological score were evaluated.The expression of a-SMA,collagen type Ⅰ and Ⅲ,IL-10 in pancreatic tissue were detected by ELASA.The expression of apoptosis genes BNIP3 in pancreatic tissue was assayed by qRT-PCR.The protein of TGF-pl,Smad2,Smad3 and Smad4 in TGF-β/Smad signal pathway in pancreatic tissue was determined by Western-blot method.The culture system of PSC was divided into 2 groups with 5 in each:control group and treatment group.The expression of a-SMA,collagen type Ⅰ and Ⅲ,IL-10 for PSC were detected by ELASA.The expression of apoptosis genes BNIP3 for PSC was assayed by qRT-PCR.The protein of TGF-βl,Smad2,Smad3 and Smad4 in TGF-β/Smad signal pathway for PSC was determined by Westem-blot method.Results(1)The pancreatic fibrosis and pathological score in treatment group were lower than model group and sham treatment group(P<0.05).The expression of α-SMA,collagen type Ⅰ and Ⅲ in tissues were less in treatment group compared with model group and sham treatment group(P<0.05)while the level of BNIP3 and IL-10 in pancreatic tissue were higher in treatment group compared with model group and sham treatment group with significant group(P<0.05).The level of TGF-βl,Smad2,Smad3 and Smad4 were lower in treatment group compared with model group and sham treatment group(P<0.05).(2)The expression of a-SMA,collagen type I and III were less in treatment group compared with control group(P<0.05).The level of BNIP3 and IL-10 were higher in treatment group compared(P<0.05).The level of TGF-βl,Smad2,Smad3 and Smad4 were less in treatment group compared with control group(P<0.05).Conclusion BMSCs reduce the activation and proliferation of PSC and then lower pancreatic fibrosis degree in rats with chronic pancreatitis.