Effect of <italic style="font-style: italic">Bushen Jianpi Formula </italic>（补肾健脾方） Drug Containing Serum on Dendritic Cell Subsets after Hepatitis B Virus Infection

LIU Ying; ZHANG Chunlei; SHAN Yawei; CHEN Xudong; WANG Lei

doi:10.13288/j.11-2166/r.2023.01.011

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Effect of Bushen Jianpi Formula （补肾健脾方） Drug Containing Serum on Dendritic Cell Subsets after Hepatitis B Virus Infection

更新时间：2022-12-29

- Effect of Bushen Jianpi Formula （补肾健脾方） Drug Containing Serum on Dendritic Cell Subsets after Hepatitis B Virus Infection
- Journal of Traditional Chinese Medicine Vol. 64, Issue 1, Pages: 55-61(2023)
- 作者机构：
  
  1.上海中医药大学附属龙华医院，上海市宛平南路725号，200032
  2.上海中医药大学护理学院
- 作者简介：
- 基金信息：
- DOI：10.13288/j.11-2166/r.2023.01.011
  CLC：
- Received：04 April 2022，
  
  Revised：2022-10-01，
  
  Published：02 January 2023
- 稿件说明：
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刘影,张春蕾,单亚维等.补肾健脾方含药血清对乙型肝炎病毒感染后树突状细胞亚群的影响[J].中医杂志,2023,64(01):55-61.

LIU Ying,ZHANG Chunlei,SHAN Yawei,et al.Effect of Bushen Jianpi Formula （补肾健脾方） Drug Containing Serum on Dendritic Cell Subsets after Hepatitis B Virus Infection[J].Journal of Traditional Chinese Medicine,2023,64(01):55-61.
刘影,张春蕾,单亚维等.补肾健脾方含药血清对乙型肝炎病毒感染后树突状细胞亚群的影响[J].中医杂志,2023,64(01):55-61. DOI： 10.13288/j.11-2166/r.2023.01.011.

LIU Ying,ZHANG Chunlei,SHAN Yawei,et al.Effect of Bushen Jianpi Formula （补肾健脾方） Drug Containing Serum on Dendritic Cell Subsets after Hepatitis B Virus Infection[J].Journal of Traditional Chinese Medicine,2023,64(01):55-61. DOI： 10.13288/j.11-2166/r.2023.01.011.

摘要

目的

探讨补肾健脾方对不同载量乙型肝炎病毒（HBV）感染后树突状细胞（DC）亚群抑制状态的影响。

方法

分离健康人外周血单个核细胞（PBMC），采用SD大鼠制备空白及补肾健脾方含药血清，采用HepG 2.2.15细胞株制备HBV病毒液。分为低病毒刺激（病毒浓度为1×10

IU/孔）和高病毒刺激（病毒浓度为1×10

IU/孔）实验。低病毒刺激实验每组设6个复孔，每孔细胞为1×10

，分为正常组一、低病毒组（病毒浓缩液1 μl）、空白血清组一（病毒浓缩液1 μl+10%空白血清200 μl）、补肾健脾方含药血清组一（病毒浓缩液1 μl+10%补肾健脾方含药血清200 μl）；高病毒刺激实验每组设6个复孔，每孔细胞为1×10

，分为正常组二、高病毒组（病毒浓缩液100 μl）、空白血清组二（病毒浓缩液100 μl+10%空白血清200 μl）、补肾健脾方含药血清组二（病毒浓缩液100 μl+10%补肾健脾方含药血清200 μl）。两个实验均在干预12、24、36、48 h时采用流式细胞仪检测各组DC亚群髓样树突状细胞（mDC）和浆细胞样树突状细胞（pDC）含量。

结果

低病毒刺激实验中，补肾健脾方含药血清组一各时间mDC、pDC含量均高于同时间点空白血清组一、低病毒组（

＜0.05）；补肾健脾方含药血清组一mDC含量本组各时间点比较差异均无统计学意义（

＞0.05），pDC含量12 h、24 h时较本组36 h、48 h低（

＜0.05）。高病毒刺激实验中，补肾健脾方含药血清组二各时间mDC、pDC含量均高于同时间点空白血清组二和高病毒组（

＜0.05）；补肾健脾方含药血清组二pDC含量12 h、24 h时较本组36 h、48 h 低（

＜0.05），mDC含量36 h时较12 h、24 h低，48 h时较12 h低（

＜0.05）。不同载量HBV刺激12、24、36、48 h时，高病毒组及低病毒组mDC、pDC含量均随干预时间延长而降低（

＜0.05）；高病毒组各时间mDC含量均低于同时间点低病毒组（

＜0.05），pDC含量仅12 h时低于低病毒组（

＜0.05）。

结论

补肾健脾方含药血清可改善HBV对DC亚群的抑制，提高mDC、pDC含量进而增强机体对HBV的免疫应答。

Abstract

Objective

To investigate the regulatory effect of Bushen Jianpi Formula （补肾健脾方， BJF） on the inhibition of dendritic cells （DC） subsets after infection with different hepatitis B virus （HBV）loads.

Methods

Peripheral blood mononuclear cells （PBMC） were obtained from healthy individuals. SD rats were used to prepare blank and BJP-containing serum， and HepG 2.2.15 cell line was used to prepare HBV virus liquid. The experiments were divided into low virus stimulation experiment （concentration of 1×10

IU/well） and high virus stimulation experiment （concentration of 1×10

IU/well）. In the low virus stimulation experiments， 6 replicate wells each with 1×10

cells were set in the normal groupⅠ， low virus group （1 μl of concentrated virus fluid）， blank serum groupⅠ （1 μl of concentrated virus fluid plus 200 μl of 10% blank serum）， and BJF-containing serum groupⅠ （1 μl of concentrated virus fluid plus 200 μl of 10% BJF-containing serum）. In the high virus stimulation experiments， 6 replicate wells each with 1×10

cells were set in the normal groupⅡ， high virus group （100 μl of concentrated virus fluid）， blank serum groupⅡ （100 μl of concentrated virus fluid plus 200 μl of 10% blank serum）， and BJF-containing serum groupⅡ （100 μl of concentrated virus fluid plus 200 μl of 10% BJF-containing serum）. The content of myeloid dendritic cells （mDC） and plasmacytoid dendritic cells （pDC） of DC subsets in each group in the two experiments were detected by flow cytometry at 12 h， 24 h， 36 h and 48 h.

Results

In the low virus stimulation experiment， the contents of mDC and pDC in the BJF-containing groupⅠ were significantly higher than those in the blank serum groupⅠ and low virus stimulation group measured at the same timepoint （

＜0.05）， and were not significantly different in mDC content when compared between different timepoints in the BJF-containing groupⅠ （

＞0.05）； lower pDC measurements were found at 12 h and 24 h than at 36 h and 48 h in the other two groups （

＜0.05）. In the high virus stimulation experiment， the proportions of mDC and pDC in BJF-containing serum groupⅡ were higher than those in blank serum groupⅡ and high virus stimulation group measured at the same timepoint （

＜0.05）， and lower contents of pDC were found when measured at 12 h and 24 h than at 36 h and 48 h， and lower mDC contents at 36 h than at 12 h and 24 h， as well as at 48 h than at 12 h in the BJF-containing serum groupⅡ （

＜0.05）. The proportions of mDC and pDC in both high and low virus stimulation groups decreased with the increasing intervention time of HBV stimulation at different loadings at 12 h， 24 h， 36 h and 48 h （

＜0.05）. The proportion of mDC in the high virus stimulation group was lower than that in the low virus stimulation group at all timepoints （

＜0.05）， and pDC was lower than that in the low virus stimulation group only at 12 h （

＜0.05）.

Conclusion

BJF-containing serum can improve the inhibition of DC subsets by HBV and can increase the ratio of mDC and pDC， thus enhancing the body's immune response to HBV.

关键词

Keywords

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Effect of Bushen Jianpi Formula （补肾健脾方） Drug Containing Serum on Dendritic Cell Subsets after Hepatitis B Virus Infection

DOI：10.13288/j.11-2166/r.2023.01.011

摘要

Abstract

关键词

Keywords

references