Infusion Of Bone Marrow-Derived Mesenchymal Stem Cells After Selective Portal Vein Ligation Enhances Functional Hepatic Reserves Of Icteric Rats

Objective: obstructive jaundice (OJ) is one of the common clinical pathological statein Hepatobiliary and Pancreatic Surgery. In order to better understand its pathogenesis,scholars have tried a variety of rat obstructive jaundice models and its reversed bybiliary drainage. However, complications such as intestinal reflux and blockage of bileduct were still common. By using of microsurgical techniques, we aim to establish astable rat model with obstructive jaundice reversed by internal drainage.Methods: After a pilot study, we have developed a standardized surgical procedure.All operations are carried out under a microscope. In the first laparotomy to establishmodel of jaundice, approximately3mm of drainage tube was introduced into theduodenal side of the common bile duct (CBD) and fixed in place with a8-0prolineligature. The other end of the tube was fixed to abdominal wall. In the secondlaparotomy, the drainage tube was inserted into the markedly dilated proximal CBDand tied in position. The liver weight, body weight, biochemical indexes, ultrasoundelastography and pathology were measured and tolerance of major liver resection wasassessed.Results: After biliary drainage, the body weight began to increase, and serum glucoseand albumin concentration rapidly increased to normal levels. The nutritional statuswas significantly improved. The complications, such as reflux and bile duct blockagewere not found in this model. In a few rats, the serum amylase and lipase increased thefirst day after operation, but rapidly decreased to normal levels. After biliary drainage,biochemical indicators, liver weight to body weight ratio, value of ultrasoundelastography, liver pathological status gradually came to normal levels. The85%hepatectomy was performed in six of the rats receiving biliary drainage for seven days.All these rats survived, and the remant liver regenerate well..Conclusion: By using microsurgical techniques, we developed an improved rat model of OJ reversed by internal drainage, particularly suitable for the long-term biliarydrainage. Objective: Hilar cholangiocarcinoma (HCCA) is a tumor with high invasiveness. Toachieve tumor-free resection margin, major liver resection was often performed, withhigh risk for serious short-term complications of residual liver failure. In recent years,the bone marrow mesenchymal stem cells (BMSCs) were used for the treatment ofliver disease. In the present study, we want to explore weather selective biliarydrainage (SBD) combined with selective portal vein ligation (SPVL), followed byadditional BMSCs infusion, due to more quickly increase in functional hepaticreserves of icteric rats.Methods: BMSCs from rats were isolated and cultured in vitro by using the wholebone marrow adherence method. BMSCs surface markers were detected using flowcytometry. Osteogenic ability was examined by alizarin red staining. Adipogenicability was measured by Oil red O staining. After the jaundice models were created,the animals were randomly divided into three groups, namely SBD group,SBD+SPVL group and the SBD+SPVL+BMSCs treatment group. Three observationtime points, which were1,3and7days after the intervention, were chosen. The liverweight to body weight ratio, spleen weight to body weight ratio,99mTc-GSAradioactive counts, ICG clearance test, the serum biochemical indicators werecompared between the three groups. Tight junction protein occludin in the liver tissuewas detected by Western blot; The expression of IL-1β in the liver was detected byELISA assay; Ki-67expression in the hyperplasia liver was examined byimmunohistochemical staining.Results: The rat BMSCs were successfully obtained, and the CD90positive rate ofthe3rd generation was93.28%. These BMSCs could be induced to cells withosteogenic ability or adipogenic ability in vitro. Rats with OJ for14days, could not tolerate70%hepatectomy; the value of Young elastic modulus was significantlyhigher than that of OJ for7days. In this case, we chose to create a rat model ofobstructive jaundice of14days for further study. After SPVL, the BMSCs wereinfused through portal vein. Twenty-four hours later, the CM-Dil labeled BMSCsgathered around the blood vessels. Seven days later, the labeled cell became diffuselydistributed in the liver parenchyma, while fluorescence of membrane markers CM-Dildecreased with cell division. BMSCs accelerated the growth rate ofnon-portal-vein-ligated liver, and boost atrophy of the ligated side of liver. Theradioactive count of the lower lobe of the liver in SBD+SPVL+BMSCs group wassignificantly higher than the SBD+SPVL group on day1and3(192.3±15.8vs.158.7±23.1and274.2±10.2vs.225.3±29.8, p <0.05). Compared with SBD+SPVLgroup, the TBIL and ALP level of the SBD+SPVL+BMSCs group were much loweron the3rd and7th days after the intervention. The IL-1β concentration in hyperplasiaside liver of SBD+SPVL+BMSCs group is lower than the other two groups, especiallyon the first and third days after intervention. The Ki-67positive cells increased rapidlyin hyperplastic liver of SBD+SPVL+BMSCs group, and accounted for22±5%on thethird day after intervention. The levels of occludin in the regenerative liver ofSBD+SPVL+BMSCs group were higher on the3rd and7th day after intervention..Conclusion: SBD combined with SPVL followed by additional portal vein infusion ofBMSCs, which could improve liver function, alleviate the inflammatory response ofthe liver, repaire liver cells damage and accelerate liver regeneration, might be anefficient way to increase functional Hepatic Reserves of Icteric Rats.