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人脐带血单个核细胞联合人脐带间充质干细胞疗法对乙型肝炎肝硬化患者肝功能、炎症程度及免疫功能的影响

韩霞 云升

韩霞,云升. 人脐带血单个核细胞联合人脐带间充质干细胞疗法对乙型肝炎肝硬化患者肝功能、炎症程度及免疫功能的影响[J]. 临床肝胆病杂志, 2021, 37(8): 1822-1828. DOI: 10.3969/j.issn.1001-5256.2021.08.016
引用本文: 韩霞,云升. 人脐带血单个核细胞联合人脐带间充质干细胞疗法对乙型肝炎肝硬化患者肝功能、炎症程度及免疫功能的影响[J]. 临床肝胆病杂志, 2021, 37(8): 1822-1828. DOI: 10.3969/j.issn.1001-5256.2021.08.016
HAN X, YUN S. Effect of combined treatment with human umbilical cord blood mononuclear cells and human umbilical cord mesenchymal stem cells on liver function, inflammation grade, and immune function in patients with hepatitis B cirrhosis[J]. J Clin Hepatol, 2021, 37(8): 1822-1828. DOI: 10.3969/j.issn.1001-5256.2021.08.016
Citation: HAN X, YUN S. Effect of combined treatment with human umbilical cord blood mononuclear cells and human umbilical cord mesenchymal stem cells on liver function, inflammation grade, and immune function in patients with hepatitis B cirrhosis[J]. J Clin Hepatol, 2021, 37(8): 1822-1828. DOI: 10.3969/j.issn.1001-5256.2021.08.016

人脐带血单个核细胞联合人脐带间充质干细胞疗法对乙型肝炎肝硬化患者肝功能、炎症程度及免疫功能的影响

DOI: 10.3969/j.issn.1001-5256.2021.08.016
基金项目: 

内蒙古医科大学科技百万工程联合资助项目 YKD2017KJBW〔LH〕05

内蒙古自治区科技厅资助项目 201802150

详细信息
    通讯作者:

    云升,yunsheng_@163.com

  • 中图分类号: R512.62;R575.2

Effect of combined treatment with human umbilical cord blood mononuclear cells and human umbilical cord mesenchymal stem cells on liver function, inflammation grade, and immune function in patients with hepatitis B cirrhosis

Funds: 

Inner Mongolia Medical University Science and Technology One Million Engineering Joint Joint Venture Project YKD2017KJBW〔LH〕05

Inner Mongolia Autonomous Region Science and Technology Department Funded Project 201802150

  • 摘要:   目的  探讨人脐带血单个核细胞(hUCB-MNCs)联合人脐带间充质干细胞(hUC-MSCs)对乙型肝炎相关失代偿期肝硬化患者的肝功能、炎症程度及免疫功能的影响。  方法  选取2016年11月—2019年6月在内蒙古医科大学附属医院诊治的11例肝硬化患者,所纳入患者第1周输注1次hUCB-MNCs(>18×109/次),第2、3、4周每周输注1次hUC-MSCs,每次输注1×106/kg,在治疗结束的第4、8、12周进行复查,对比治疗前后肝功能、血氨、凝血因子、血清细胞因子及淋巴细胞亚群的变化,同时观察记录神经、精神症状的改变。计量资料组间比较采用单因素重复测量方差分析。  结果  hUCB-MNCs联合hUC-MSCs治疗后的11例患者其精神、神经症状与细胞输注前相比较有所改善;肝功能各项指标及凝血功能基本趋于正常(P值均<0.05);细胞联合输注后的第12周,血氨水平显著下降(P<0.05);炎性细胞因子IL-6、TNFα水平降低(F值分别为49.497、37.071,P值均<0.05);而抗炎细胞因子TGFβ和IL-10水平显著上升(F值分别为35.843、15.918,P值均<0.05);同时CD3+CD8+T淋巴细胞和CD19+B淋巴细胞均降低(F值分别为52.242、89.097,P值均<0.05);免疫调节性T淋巴细胞CD4+CD25+Treg明显升高(F=17.337,P<0.05)。  结论  hUCB-MNCs联合hUC-MSCs治疗肝硬化,与细胞输注前相比较,可改善肝功能,减少炎性细胞因子产生,减轻肝脏炎症反应以及肝细胞的破坏,升高免疫调节性T淋巴细胞,从而影响其自身免疫功能。

     

  • 图  1  hUC-MSCs形态(×100)

    注:a,P3代hUC-MSCs培养1 d,细胞为短杆状或梭形;b,培养第3天,细胞呈梭形,巢状或单极辐射生长。

    图  2  hUC-MSCs表面标记表达

    注:a,阴性对照组;b,CD44和CD105表达为98.59%;c,CD44和CD90表达为99.06%;d,CD34和HLA-DR表达为0.08%;e,山东脐血库CD34表达为1.66%。

    图  3  hUC-MSCs分化诱导的肝细胞及神经细胞形态(×400)

    注:a,hUC-MSCs分化诱导的肝细胞;b,hUC-MSCs分化诱导的神经细胞。

    表  1  入组患者的信息和条件

    患者 年龄(岁) 性别 腹水 TBil(μmol/L) Alb(g/L) PT(s) Child-Pugh分级
    No.1 50 - 48.1 26.5 19.3 B级
    No.2 48 - 47.9 22.9 19.9 B级
    No.3 55 47.4 24.8 22.5 B级
    No.4 56 - 34.9 26.0 22.9 B级
    No.5 60 54.4 22.8 20.5 C级
    No.6 52 - 48.5 31.9 20.2 B级
    No.7 41 - 43.8 40.8 19.4 B级
    No.8 49 - 50.2 23.9 17.2 B级
    No.9 51 34.8 27.4 16.2 B级
    No.10 53 67.6 22.6 19.4 C级
    No.11 51 48.7 21.9 19.4 B级
    下载: 导出CSV

    表  2  细胞输注前后肝功能变化

    输注时间 ALT(U/L) AST(U/L) ALP(U/L) GGT(U/L) TBil(μmol/L) Alb(g/L) PTA(%) PT(s)
    输注前 88.4±15.31) 99.8±25.01) 123.8±39.41) 305.1±83.41) 47.8±8.11) 26.5±6.41) 45.8±7.91) 16.2±1.31)
    输注(4周) 77.9±22.7 85.5±31.3 97.5±40.1 189.1±66.4 23.2±9.8 30.5±3.6 66.0±6.0 12.6±0.5
    输注(8周) 59.1±21.5 26.8±7.9 58.1±17.6 86.6±39.9 9.9±5.3 36.0±4.2 74.9±6.9 12.2±1.8
    输注(12周) 42.7±12.3 24.2±8.3 59.5±28.0 43.2±19.8 6.7±1.8 40.8±2.4 87.6±3.0 11.3±0.2
    F 14.496 41.163 10.941 45.740 79.730 20.496 86.289 1 029.600
    P <0.05 <0.05 <0.05 <0.05 <0.05 <0.05 <0.05 <0.05
    注:与输注后的第4、8、12周比较, 1)P<0.05。
    下载: 导出CSV

    表  3  hUCB-MNCs联合hUC-MSCs输注前后血氨值变化

    输注时间 血氨(μmol/L)
    输注前 166.6±47.71)
    输注(4周) 115.6±32.8
    输注(8周) 87.2±36.3
    输注(12周) 66.2±32.5
    F 104.6
    P P<0.05
    注:与输注后的第4、8、12周比较, 1)P<0.05。
    下载: 导出CSV

    表  4  hUCB-MNCs联合hUC-MSCs输注前后Child-Pugh评分变化

    输注时间 Child-Pugh评分
    输注前 10.10±2.431)
    输注(4周) 8.36±2.94
    输注(8周) 6.82±2.27
    输注(12周) 6.09±1.58
    F 68.4
    P P<0.05
    注:与输注后的第4、8、12周比较, 1)P<0.05。
    下载: 导出CSV

    表  5  hUCB-MNCs联合HUC-MSCs输注前后细胞因子和淋巴细胞亚群变化

    输注时间 IL-6(ng/L) TNFα(ng/L) IL-10(pg/L) TGFβ(pg/L) CD3+ CD3+CD4+ CD3+CD8+ CD4+CD25+ CD19+
    细胞输注前 198.8±54.01) 300.6±73.51) 18.3±7.81) 25.9±4.31) 75.8±11.01) 23.3±4.21) 48.3±9.31) 1.3±1.21) 32.1±4.21)
    输注后(4周) 96.2±19.6 198.9±106.4 48.9±15.7 49.3±7.1 74.8±8.9 53.1±10.9 20.4±6.8 18.3±9.5 12.7±3.6
    输注后(8周) 58.9±16.9 92.2±26.1 47.3±8.7 43.9±4.4 76.9±12.4 54.0±7.7 20.4±3.4 18.0±5.0 11.7±5.3
    输注后(12周) 59.8±16.4 92.8±13.7 49.6±15.9 35.5±6.4 80.1±10.9 55.9±6.4 16.7±6.0 19.1±8.3 6.1±2.4
    F 49.497 37.071 15.918 35.843 0.493 44.755 52.242 17.337 89.097
    P <0.01 <0.01 <0.01 <0.01 >0.05 <0.01 <0.01 <0.01 <0.01
    注:与输注后的第4、8、12周比较, 1)P<0.05。
    下载: 导出CSV
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