大鼠骨髓mMSC和RS细胞肝组织内注射对肝硬化大鼠肝功能及肝脏病变的改善作用观察

Improvement of liver function and alleviation of liver lesions in cirrhotic rats by intrahepatic injection of rat bone marrow mature mesenchymal stem cells and rapidly self-renewing cells

目的观察肝组织内注射大鼠成熟间充质干细胞(mature MSC,mMSC)与快速自我更新细胞(rapidly selfrenewing cell,RS细胞)对肝硬化大鼠的治疗效果。方法取2周龄Wistar雄性大鼠1只,处死后取胫骨、股骨,分离、培养、鉴定大鼠骨髓mMSC、RS细胞。取36只Wistar大鼠制备肝硬化模型,造模成功后随机分为A、B、C三组,A、B组分别肝组织内注射mMSC、RS细胞,C组肝组织内注射NS。分别于造模成功和治疗1、2、4周时,各组尾静脉取血检测血清谷丙转氨酶(ALT)、总胆红素(TBIL)。治疗4周时处死三组大鼠,观察肝脏形态,对肝组织进行病理分级评分(包括肝纤维化、肝细胞坏死及脂肪变性评分)。结果与C组比较,治疗治疗1周、2周、4周时A及B组血清ALT、TBIL水平降低(P均<0.05)。A组大鼠肝纤维化、肝细胞坏死、脂肪变性评分分别为(2.42±0.51)、(1.92±0.51)、(1.83±0.39)分,病理评分为(6.25±0.87)分;B组分别为(2.17±0.39)、(1.58±0.51)、(1.75±0.45)及(5.50±0.80)分;C组分别为(3.67±0.49)、(3.17±0.39)、(1.83±0.58)及(8.75±1.14)分,与C组比较,A及B组肝纤维化、肝细胞变性坏死评分低(P均<0.05)。C组形成肝脏假小叶形成,肝细胞水肿变性、坏死,可见少量脂肪空泡;A、B组肝脏假小叶的纤维纤细,肝细胞变性、坏死程度轻,少量脂肪空泡。结论肝组织内注射大鼠mMSC、RS细胞的肝硬化大鼠血清ALT、TBIL水平降低,肝纤维化及肝细胞变性程度减轻。

Objective rapidly self-renewing cells(RS cells)into liver tissues on cirrhotic rats.Methods killed,and then the tibia and femur were taken to isolate,culture and identify mMSC and RS cells in rat bone marrow.Thirty-six Wistar rats were selected to make liver cirrhosis models.After successful modeling,they were randomly divided into three groups:groups A,B and C.The mMSC and RS cells were injected into the liver tissues of rats in the groups A and B,respectively,and NS was injected into the liver tissues of rats in the group C.After successful modeling and 1,2,and 4 weeks of treatment,blood samples were taken from tail vein of each group to detect serum alanine aminotransferase(ALT)and total bilirubin(TBIL).At 4 weeks after treatment,rats of three groups were killed,the liver morphology was observed,and the liver tissues was scored by pathological grading(including liver fibrosis,hepatocyte necrosis,and steatosis).Results and 4 weeks after treatment(all P<0.05).In the group A,the scores of hepatic fibrosis,hepatocyte necrosis and steatosis were(2.42±0.51),(1.92±0.51)and(1.83±0.39)points,respectively,and the pathological score was(6.25±0.87)points.The scores in the group B were(2.17±0.39),(1.58±0.51),(1.75±0.45)and(5.50±0.80)points,respectively.The scores in the group C were(3.67±0.49),(3.17±0.39),(1.83±0.58)and(8.75±1.14)points,respectively.Compared with the group C,the scores of hepatic fibrosis and hepatocyte degeneration and necrosis in the groups A and B were lower(all P<0.05).In the group C,liver pseudolobules were formed,and edema,degeneration and necrosis were found in the liver cells,and a small amount of fat vacuoles could be seen.In the groups A and B,the fibers of liver pseudolobules were slender,the degree of degeneration and necrosis of liver cells were lighter,and a few fat vacuoles were found.Conclusion liver tissues,and the degree of liver fibrosis and hepatocyte degeneration decrease.