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  

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

作　　者：刘影 张春蕾 单亚维[2] 陈旭东[1] 王磊[1] LIU Ying;ZHANG Chunlei;SHAN Yawei;CHEN Xudong;WANG Lei(Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Mediceine,Shanghai,200032;School of Nursing,Shanghai University of Traditional Chinese medicine)

机构地区：[1]上海中医药大学附属龙华医院,上海市200032 [2]上海中医药大学护理学院

出　　处：《中医杂志》2023年第1期55-61,共7页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81973725)。

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^(5)IU/孔)和高病毒刺激(病毒浓度为1×107IU/孔)实验。低病毒刺激实验每组设6个复孔,每孔细胞为1×10^(6),分为正常组一、低病毒组(病毒浓缩液1μl)、空白血清组一(病毒浓缩液1μl+10%空白血清200μl)、补肾健脾方含药血清组一(病毒浓缩液1μl+10%补肾健脾方含药血清200μl);高病毒刺激实验每组设6个复孔,每孔细胞为1×10^(6),分为正常组二、高病毒组(病毒浓缩液100μl)、空白血清组二(病毒浓缩液100μl+10%空白血清200μl)、补肾健脾方含药血清组二(病毒浓缩液100μl+10%补肾健脾方含药血清200μl)。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(P<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(P>0.05),pDC含量12 h、24 h时较本组36 h、48 h低(P<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(P<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(P<0.05),mDC含量36 h时较12 h、24 h低,48 h时较12 h低(P<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(P<0.05);高病毒组各时间mDC含量均低于同时间点低病毒组(P<0.05),pDC含量仅12 h时低于低病毒组(P<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、pDCObjective 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 1×10~5 IU/well)and high virus stimulation experiment(concentration of 1×10~7 IU/well).In the low virus stimulation experiments,6 replicate wells each with 1×10~6 cells were set in the normal groupⅠ,low virus group(1μl of concentrated virus fluid),blank serum groupⅠ(1μl of concentrated virus fluid plus 200μl of 10%blank serum),and BJF-containing serum groupⅠ(1μl of concentrated virus fluid plus 200μl of 10%BJF-containing serum).In the high virus stimulation experiments,6 replicate wells each with 1×10~6 cells were set in the normal groupⅡ,high virus group(100μl of concentrated virus fluid),blank serum groupⅡ(100μl of concentrated virus fluid plus 200μl of 10%blank serum),and BJF-containing serum groupⅡ(100μl of concentrated virus fluid plus 200μl 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 groupⅠwere significantly higher than those in the blank serum groupⅠand low virus stimulation group measured at the same timepoint(P<0.05),and were not significantly different in mDC content when compared between different timepoints in the BJF-containing groupⅠ(P>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(P<0.05).In the high virus stimulation experiment,the proportions of mDC and pDC in

关 键 词：乙型肝炎病毒 髓样树突状细胞 浆细胞样树突状细胞 外周血单个核细胞 补肾健脾方 

分 类 号：R285.5[医药卫生—中药学]