人ADSCs移植对大鼠急性重症胰腺炎的治疗作用及其机制

Therapeutic effect of human ADSCs transplantation on acute severe pancreatitis in rats and its mechanism

目的探讨人脂肪间充质干细胞(ADSCs)移植对大鼠急性重症胰腺炎(SAP)的治疗作用及其可能的作用机制。方法选择雄性SD大鼠45只,随机分为假手术组、模型组、移植组,每组15只。模型组、移植组按胰胆管逆行注射牛磺胆酸钠法建立SAP模型,当观察到胰腺充血、水肿时,移植组于胰腺被膜下注射人ADSCs,模型组不予处理。假手术组开腹后经胰胆管逆行注射台式液。移植6、12、24h,各组分别取5只,记录腹水量,采用ELISA法检测血清淀粉酶、TNF-α;取胰头组织,HE染色,观察胰腺组织形态学变化;分别采用RT-PCR法、Western blotting法检测胰头组织磷酸化蛋白激酶B(p-AKT)、磷酸化核因子κB(p-NF-κB)mRNA与蛋白表达。结果随着移植时间延长,移植组和模型组腹水量较假手术组均逐渐增多(P均<0.05),且移植组各时间腹水量均低于模型组,但差异无统计学意义(P均>0.05);移植组和模型组各时间血清淀粉酶、TNF-α水平均明显高于对照组,但移植组血清淀粉酶、TNF-α水平较模型组同期均明显降低(P均<0.05)。移植组胰头组织水肿、坏死及炎性细胞浸润情况均较模型组轻微。模型组p-AKT、p-NF-κBmRNA与蛋白相对表达量均明显高于假手术组,而移植组p-AKT、p-NF-κBmRNA与蛋白相对表达量均明显低于模型组(P均<0.05)。结论人ADSCs移植对大鼠SAP具有一定治疗作用,其作用机制可能与抑制AKT磷酸化进而抑制NF-κB激活,继而产生弱化“二次打击”扳机点作用有关。

Objective To investigate the therapeutic effect of human adipose-derived mesenchymal stem cells (ADSCs) transplantation on acute severe pancreatitis (SAP) in rats and its possible mechanism. Methods Forty-five male Sprague-Dawley rats were randomly divided into the sham operation group, model group and transplantation group. In the model group and the transplantation group, we used the retrograde injection of sodium taurocholate to establish the SAP models. When pancreatic congestion and edema were observed, the rats in the transplantation group were injected with human ADSCs under the pancreas, and the model group was not treated. Rats in the sham operation group were retrogradely injected through the pancreatic duct after laparotomy. Five rats in each group were taken at 6, 12, and 24 h after transplantation, and the amount of ascites was recorded;the serum amylase and TNF-α were detected by ELISA;the pancreatic head tissues were taken and stained with HE staining to observe the morphological changes of pancreas;the p-AKT and p-NF-κB mRNA and protein expression in the pancreatic head tissues were detected by RT-PCR and Western blotting, respectively. Results Over the time of transplantation, the ascites volume in the transplantation group and the model group increased gradually (P<0.05), and the ascites volume in the transplantation group was lower than that in the model group, but the difference was not statistically significant (P>0.05). The serum amylase and TNF-α levels in the transplantation group and the model group were higher than those in the control group at each time, but the serum amylase and TNF-α levels in the transplantation group were significantly lower than those in the model group (all P<0.05). Morphological observation of the head of the pancreas showed that the edema, necrosis and inflammatory cell infiltration of the head tissues of the transplantation group were slightly milder than those of the model group. The relative mRNA and protein expression levels of p-AKT and p-NF-κB in the model group were significantly higher than those in the sham operation group, while the relative mRNA and protein expression levels of p-AKT and p-NF-κB in the transplantation group were significantly lower than those in the model group (all P<0.05). Conclusion Human ADSCs transplantation has a certain therapeutic effect on rat SAP, and its mechanism may be related to inhibition of AKT phosphorylation and inhibition of NF-κB activation, which in turn may weaken the role of "second strike" trigger point.