Effect of rhubarb-free anthraquinone in the treatment of acute pancreatitis with adipose-derived stem cell transplantation in rats / 中国组织工程研究

ABSTRACT

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 anthraquinone group and cell transplantation group (P < 0.05). (2) The results of hematoxylin-eosin staining showed that the pathological changes in the pancreatic tissue of rats were significantly reduced in the rhubarb-free anthraquinone group and cell transplantation group, such as fatty degeneration, hemorrhage, cell necrosis and inflammatory cell infiltration. The remission of pathological changes was more obvious in the combined treatment group. (3) The number of positive cells labeled by PKH-26 was the highest in the combined treatment group, followed by the cell transplantation group, while there were no PKH-26-positive cells in the rhubarb-free anthraquinone group and model group (P < 0.05). (4) Compared with the model group, the number of apoptotic cells in the pancreatic tissue was significantly reduced in the rhubarb-free anthraquinone group and cell transplantation group, and lowest in the combined treatment group (P < 0.01). (5) The expression of transforming growth factor-β1 and Smad2/3 at gene and protein levels was highest in the model group, followed by the rhubarb-free anthraquinone group and cell transplantation group (P < 0.05) , and lowest in the combined treatment group (P < 0.01). To conclude, the combined intervention of rhubarb-free anthraquinone and adipose-derived stem cell transplantation could effectively improve the blood biochemical index level in the rats with acute pancreatitis, significantly relieve the degree of inflammatory response, pancreatic tissue morphology, and apoptosis of pancreatic cells, which may be related to the reduction of the transforming growth factor-β/Smad signal pathway.