B 型超声引导下肝包膜下骨髓间充质干细胞移植治疗大鼠肝损伤

摘要

目的探讨在 B 型超声引导下经皮向肝包膜下注射移植骨髓间充质干细胞（MSC）对大鼠肝损伤的治疗作用。方法四氯化碳（CCl4）皮下注射建立肝损伤大鼠模型。首次移植：手术开腹，在肝损伤大鼠肝包膜下注入1 mL 含MSC 胶原培养液（MSC 移植组）；在肝损伤大鼠和正常大鼠肝包膜下分别注入1 mL 不含 MSC 胶原培养液（分别为肝损伤对照组和正常对照组）。各组注入物均可在肝包膜下形成胶原凝胶。第2次和第3次移植：分别在首次移植后第3周和第4周在 B 超引导下经皮向上一次移植留在肝包膜下形成的凝胶囊腔内注射含 MSC 培养液（MSC 移植组）或不含 MSC 培养液（肝损伤对照组和正常对照组）。在第3次移植后1周，检测各组大鼠 Alb、ALT、TBil；称大鼠体重和肝重，计算肝指数。结果MSC 移植组血浆 Alb 水平（32．5±6．7）g/L 明显高于肝损伤对照组（24．5±7．4）g/L（P ＜0．05）；而 ALT（97．7±19．6）U/L、TBil（6．5±1．4）μmol/L 和肝指数（4．4±0．5）均明显低于肝损伤对照组的（321．8±27．4）U/L、（38．2±14．3）μmol/L 和（5．5±0．7）（均 P ＜0．05）。结论将胶原溶液注入大鼠肝包膜下可形成胶原凝胶囊腔；可在 B 型超声引导下多次经皮将 MSC 注入肝包膜下凝胶囊腔（移植到肝实质表面）；5周内3次移植 MSC 到肝包膜下可使肝损伤大鼠的肝功能得到明显改善。%Objective Bone marrow mesenchymal stem cells (MSCs)may be a useful therapeutic approach for liver disorders,but there is not a satisfactory route of transplanting MSCs into liver yet.The research was to study a method of transplanting large quantity of MSCs into liver subcapsule of rats by transcutaneous intra-subcapsule injection guided by B ultrasound and its therapeutic effect on liver injury. Methods The model rats of liver injury were established by subcutaneous injection of carbon tetrachloride (CCl4 )for 8 weeks.(1)Initial transplantation(performed on exposed liver by operation):Liver-injured rats were divided into MSCs transplant group and liver injury control group by injecting collagen solution with MSCs or without MSCs into liver subcapsule to form collagen gel cyst respectively.In addition,normal rats were injected collagen solution without MSCs,as normal control group.(2)Second and third transplantations:the initial transplantations were repeated but were performed by transcutaneous intra-gel cyst injection under guidance of B ultrasound without collagen.At 1 week after 3rd transplant,liver index and plasma levels of albumin(Alb),total bilirubin (TBil)and alanine transaminase (ALT)were examined.Results Level of Alb in MSCs transplant group was significantly higher than that in liver injury control group (P <0.05).Value of liver index and levels of ALT,TBil of tMSCs transplant group were significantly lower than those of liver injury control group (P < 0.05 ).Conclusion MSCs could be transplanted into subcapsule of liver in large quantity by the transcutaneous intra-gel cyst injection guided by B ultrasound.This method could improve liver functions of liver-injured rats.