ANALYSIS OF T CELL CLONALITY BY CDR3 SIZE OF T-CELL ANTIGEN RECEPTOR Vβ REPERTOIRE IN HCL AND c-ALL

Objective: To analyze the distribution and clonality of TCR Vβ subfamily T cells in hairy cell leukemia (HCL) and common-acute lymphoblastic leukemia (c-ALL). Methods: Peripheral blood mononuclear cell samples from 3 cases of HCL and 1 case of c-ALL were investigated for analysis of complementarity determining region 3 (CDR3) size of T cell receptor Vβ repertoire using reverse transcriptase-polymerase chain reaction (RT-PCR). The products were further analyzed by genescan to identify T cell clonality. Results: Some Vβ subfamily PCR products from 4 patients contained monopeak (monoclone) or a dominant peak (oligoclone). In contrast, multipeak (polyclone) distributions were found in all Vβ subfamily PCR products from normal control cases. Conclusion: T cell clonal expansion may be found in HCL and c-ALL cases that may indicate a host response directed against leukemia related antigen. In addition, it may be useful to detect the minimal residual disease.

Clonality analysis of neuroendocrine cells in gastric adenocarcinoma

AIM:To achieve a better understanding of the origination of neuroendocrine(NE)cells in gastric adenocarcinoma.METHODS:In this study,120 cases of gastric adenocarcinoma were obtained.First,frozen section-immunohistochemistrical samples were selected from a large quantity of neuroendocrine cells.Second,laser capture microdissection was used to get target cells from gastric adenocarcinoma and whole genome amplification was applied to get a large quantity of DNA for further study.Third,genome-wide microsatellite abnormalities[microsatellite instability(MSI),loss of heterozygosity (LOH)]and p53 mutation were detected by polymerase chain reaction(PCR)-single-strand conformation polymer-phism-silver staining and PCR-sequencing in order to identify the clonality of NE cells.RESULTS:The total incidence rate of MSI was 27.4%,while LOH was 17.9%.Ten cases had a highest concordance for the two types of cells.The other samples had similar microsatellite changes,except for cases 7 and10.Concordant p53 mutations exhibited in sample 4,14,21 and 27,and there were different mutations between two kinds of cells in case 7.In case 17,mutation took place only in adenocarcinoma cells.p53 mutation was closely related with degree of differentiation,tumor-node-metastasis stage,vessel invasion and lymph node metastasis.In brief,NE and adenocarcinoma cells showed the same MSI,LOH or p53 mutation in most cases(27/30).In the other three cases,different MSI,LOH or p53 mutation occurred.CONCLUSION:NE and the gastric adenocarcinoma cells may mainly derive from the same stem cells,but the remaining cases showing different origin needs further investigation.

Next-generation sequencing traces human induced pluripotent stem cell lines clonally generated from heterogeneous cancer tissue

AIM To investigate genotype variation among induced pluripotent stem cell(iPSC) lines that were clonally generated from heterogeneous colon cancer tissues using next-generation sequencing. METHODS Human iPSC lines were clonally established by selecting independent single colonies expanded from heterogeneous primary cells of S-shaped colon cancer tissues by retroviral gene transfer(OCT3/4, SOX2, and KLF4). The ten iPSC lines, their starting cancer tissues, and the matched adjacent non-cancerous tissues were analyzed using nextgeneration sequencing and bioinformatics analysis using the human reference genome hg19. Non-synonymous single-nucleotide variants(SNVs)(missense, nonsense,and read-through) were identified within the target region of 612 genes related to cancer and the human kinome. All SNVs were annotated using dbS NP135, CCDS, RefSeq, GENCODE, and 1000 Genomes. The SNVs of the iPSC lines were compared with the genotypes of the cancerous and non-cancerous tissues. The putative genotypes were validated using allelic depth and genotype quality. For final confirmation, mutated genotypes were manually curated using the Integrative Genomics Viewer. RESULTS In eight of the ten iPSC lines, one or two non-synonymous SNVs in EIF2AK2, TTN, ULK4, TSSK1 B, FLT4, STK19, STK31, TRRAP, WNK1, PLK1 or PIK3R5 were identified as novel SNVs and were not identical to the genotypes found in the cancer and non-cancerous tissues. This result suggests that the SNVs were de novo or pre-existing mutations that originated from minor populations, such as multifocal pre-cancer(stem) cells or pre-metastatic cancer cells from multiple, different clonal evolutions, present within the heterogeneous cancer tissue. The genotypes of all ten iPSC lines were different from the mutated ERBB2 and MKNK2 genotypes of the cancer tissues and were identical to those of the noncancerous tissues and that found in the human reference genome hg19. Furthermore, two of the ten iPSC lines did not have any confirmed mutated genotypes, despite being derived from cancerous tissue. These results suggest that the traceability and preference of the starting single cells being derived from pre-cancer(stem) cells, stroma cells such as cancer-associated fibroblasts, and immune cells that co-existed in the tissues along with the mature cancer cells.CONCLUSION The genotypes of iPSC lines derived from heterogeneous cancer tissues can provide information on the type of starting cell that the iPSC line was generated from.

Liver cell adenoma:A case report with clonal analysis and literature review

We report a case of liver cell adenoma （LCA） in a 33-year-old female patient with special respect to its clonality status, pathogenic factors and differential diagnosis. The case was examined by histopathology, immunohistochemistry and a clonality assay based on X-chromosomal inactivation mosaicism in female somatic tissues and polymorphism at androgen receptor focus. The clinicopathological features of the reported cases from China and other countries were compared. The lesion was spherical, sizing 2 cm in its maximal dimension. Histologically, it was composed of cells arranged in cords, most of which were two-cell-thick and separated by sinusoids. Focal fatty change and excessive glycogen storage were observed. The tumor cells were round or polygonal in shape, resembling the surrounding parenchymal cells. Mitosis was not found. No portal tract, central vein or ductule was found within the lesion. The tumor tissue showed a positive reaction for cytokeratin （CK） 18, but not for CK19, vimentin, estrogen and progesterone receptors. Monoclonality was demonstrated for the lesion, confirming the diagnosis of an LCA. Clonality analysis is helpful for its distinction from focal nodular hyperplasia.K

Characterization of mouse clonal mesenchymal stem cell lines established by subfractionation culturing method

AIM:To characterize single-cell-derived mouse clonal mesenchymal stem cells (mcMSCs) established with bone marrow samples from three different mouse strains. METHODS:We established mcMSC lines using subfractionation culturing method from bone marrow samples obtained from long bones.These lines were characterized by measuring cell growth, cell surface epitopes, differentiation potential, lineage-specific gene expression and T-cell suppression capability. Nonclonal MSCs isolated by the conventional gradient centrifugation method were used as controls. RESULTS:All mcMSC lines showed typical nonclonal MSC-like spindle shape morphology. Lines differed inoptimal growth density requirement.Cell surface epitope prof iles of these mcMSC lines were similar to those of nonclonal MSCs. However, some lines exhibited different expression levels in a few epitopes, such as CD44 and CD105. Differentiation assays showed that 90% of the mcMSC lines were capable of differentiating into adipogenic and/or chondrogenic lineages, but only 20% showed osteogenic lineage differentiation. T-cell suppression analysis showed that 75% of the lines exhibited T-cell suppression capability. CONCLUSION:mcMSC lines have similar cell morphology and cell growth rate but exhibit variations in their cell surface epitopes, differentiation potential, lineage-specifi c gene expression and T-cell suppression capability.

CLONAL PROLIFERATION AND LONG-TERM CULTURE OF MALIGNANT LYMPHOMA CELLS UNDER SERUM-FREE CULTURE CONDITIONS

We developed a serum-free culture system that promoted the growth of B cell colonies in peripheral blood, bone marrow, lymph nodes and cerebrospinal fluid (CSF) from 7 out of 8 patients with non-Hodgkin's lymphomas of B cell type. The culture cells were pretreated with or without galactose oxi-dase (GO) prior to plating. Colony growth was best supported with BCGF. A moderate increment was observed with rIL-3, as well as rIL-1β and even to a lesser degree, by rlL-2, while B cell stimulating factor-2 (rBCSF-2) and rlL-1β did not show significant activity. rGM-CSF and rG-CSF had little effect, while rM-CSF enhanced the formation of lymphoma colonies. The cells from different patients had different requirements for Staphylococcus aureus protein A and GO pretreatment. It reflected the differences in activation and differentiation status and surface properties of lymphoma cells from different patients. The cells from CSF of one patient were successfully maintained in serum-free culture medium supplemented with 10% BCGF or 5% PHA-LCM for more than 4 months. The long-term culture cells were EBV negative, phenotypically consistent with B cells and gene rearrangements for JH, Kappa and myc. This serum-free culture system allowed extensive analysis of the growth requirements for clonogenic precursors.

INFLUENCE OF IMMUNE STATUS OF THE IMMUNE DEFICIENT MICE ON THE METASTATIC PHENOTYPES OF THE HETEROGENEOUS CLONAL SUBLINES OF HUMAN LUNG GIANT CELL CARCINOMA

By using cell cloning technique, 4 sublines (A,C,D,E) were isolated from a cell line of human lung giant cell carcinoma (PLA-801). After subcutaneous inoculation in T-cell deficient BALB/c nude mice, the incidence of tumor growth and spontaneous metastasis were the highest in subline D, moderate in sublines A and E, and lowest in subline C. Tumor cells of subline C also showed similar low tumorigenicity in another T-cell deficient 615/ PB1 nude mice.However, in 615/PB1 beige nude mice with con-genitally combined immune-deficiency in both T and NK cell activity, tumor cells of the rarely metastatic subline C do produce significantly high frequency of tumor growth and spontaneous metastasis.Morphological studies (light microscope, electron microscope and immunohistochemistry) showed rich microfilaments and Vimentin positive in the cytoplasm of metastatic tumor cells. This may imply a possibility that tumor cells differentiate towards the direction favourable to spreading and metastasis.

T-ALL及T细胞株相关TCR Vβ基因谱系和克隆性分析

目的 了解T细胞 急性淋巴细胞白血病 (T ALL)患者外周血中的T细胞及T细胞株的TCRVβ基因谱系及其克隆性增殖情况。方法 利用RT PCR方法扩增 6个不同的T细胞株和 6例初发未治T ALL病人外周血单个核细胞中 2 4个TCRVβ基因的互补决定区 3(CDR3) ,PCR产物进一步经荧光标记和基因扫描分析CDR3长度而确定T细胞的克隆性 ,部分T细胞株的单克隆PCR产物进一步进行序列分析。结果 与正常人外周血表达全部 2 4个Vβ亚家族不同 ,6例T ALL病人分别表达 5～ 12个Vβ亚家族。 6例病人均存在 1个或多个Vβ亚家族的寡克隆或双克隆增殖T细胞 ,另外 ,还有一些Vβ亚家族多克隆模式发生改变 ,呈现寡克隆性增殖的趋势。T细胞株多显示为表达一个Vβ亚家族的单克隆T细胞 ,不同T细胞株的CDR3长度和序列不尽相同。结论 T ALL患者外周血T细胞的TCRVβ谱系出现限制性改变 ,均可检测到克隆性增殖T细胞 ,尚需进一步鉴定其性质 (肿瘤性或抗原特异性增殖 ) 。

用RT-PCR和Genescan分析CML急变期病人TCRVβT细胞的表达和克隆性

目的：了解CML急变期的TCRVβ亚家族T细胞的表达及其克隆性。方法：采用RT－PCR扩增4例CML急变期病人的外周血单个核细胞的TCRVβ24个亚家族的互补决定区3（CDR3），产物进一步经基因扫描分析确定T细胞的克隆性。结果：病人外周血仅表达4～9个Vβ亚家族T细胞，主要为Vβ1，Vβ2，Vβ3，Vβ5，Vβ15和Vβ17；基因扫描分析显示2例CML急粒变的部分产物为寡克隆性，而2例CML急淋变者的产物均为多克隆性。结论：CML急变期外周血仅选择性表达部分Vβ亚家族T细胞；CML急粒变存在克隆性增殖T细胞，这可能是机体对白血病细胞相关抗原的一种直接反应。该方法可用于检测微小残留病变和判断疾病复发。

抑癌基因Kmt2c杂合缺失对小鼠造血系统的影响

目的:探究组蛋白甲基转移酶Kmt2c基因杂合缺失对小鼠血液系统的影响。方法:使用CRISPR/Cas9技术构建Kmt2c基因杂合缺失的模型小鼠,血常规连续监测小鼠全血细胞计数的变化;通过体外集落形成实验探究骨髓细胞的克隆性扩增能力;流式细胞术分析突变小鼠体内原始造血细胞群(长期造血干细胞、短期造血干细胞、多能祖细胞)的比例变化。结果:成功构建Kmt2c基因杂合缺失小鼠(Kmt2c^(+/-))模型,其Kmt2c mRNA表达水平是C57BL/6J小鼠的28%。Kmt2c^(+/-)小鼠骨髓细胞体外集落形成能力随着传代次数的增加而增强,并在第四代时集落数显著高于对照组(P<0.05)。Kmt2c^(+/-)小鼠原始造血细胞群中的长期造血干细胞和短期造血干细胞比例分别为19.6%±3.3%及28.9%±4.9%较对照组的16.9%±2.6%及18.9%±2.5%有增高趋势,但差异无统计学意义(P>0.05)。Kmt2c^(+/-)小鼠的白细胞计数在监测的第12周后逐渐上升在第14周时为(9.8±1.0)×10^(9)/L,显著高于对照组的(7.3±1.4)×10^(9)/L(P<0.05)。结论:Kmt2c^(+/-)小鼠的骨髓细胞具有克隆性扩增的潜能。

山丹丹新品种‘延丹1号’的选育

以山丹丹和兰州百合为亲本,培育山丹丹新品种。通过建立山丹丹和兰州百合无性系,诱导愈伤组织。经过原生质体电融合,细胞培养再分化获得的变异植株。通过籽球培养,室内春化、室外栽培等确定变异植株性状。变异植株为无性繁殖,植株长势强壮;花色为橘红色,鳞片环抱紧密,抗旱、抗寒性强,耐贫瘠,无明显病虫害。通过在延安市宝塔区、子长市高柏山、宜川县集义镇连续3年以上测试,发现品种稳定性好、异株率低、复花能力强。经过简化基因组分析‘延丹1号’含有山丹丹和兰州百合的基因,确定通过细胞融合培育获得了山丹丹新品种。

RNA干扰下调PPP2R5C对T-ALL TCR Vβ亚家族分布和克隆增殖的影响

目的:基于前期利用靶向PPP2R5C基因的小干扰RNA(PPP2R5C-siRNA)抑制白血病T细胞增殖的结果,进一步了解PPP2R5C-siRNA对T-ALL样本中TCR Vβ亚家族基因谱系及其克隆性增殖情况.方法:利用PPP2R5C-siRNA转染2例初发未治疗T-ALL病人外周血单个核细胞(PBMC),设无关序列对照组(SC)和空白对照组(NC);利用实时定量PCR检测PPP2R5C基因表达水平.利用RT-PCR扩增各组样本中24个TCR Vβ亚家族基因的互补决定区3(CDR3);阳性产物进一步经基因扫描分析CDR3长度,了解其Vβ亚家族T细胞克隆性.结果:PPP2R5C-siRNA处理后明显下调PBMC中PPP2R5C基因表达水平.2例初发T-ALL外周血中分别可检测到10和17个Vβ亚家族,多数Vβ亚家族T细胞为多克隆性,病例1中Vβ9和Vβ17为寡克隆性,而病例2中,Vβ14和Vβ16呈寡克隆性;经过RNA干扰处理后,TCR Vβ亚家族表达率降低,仅检测到3和4个Vβ亚家族,分别为Vβ2,Vβ3,Vβ16和Vβ17.其中2个Vβ家族的寡克隆性模式发生改变,病例1中,Vβ17转为多克隆,而病例2中,Vβ16也转变为多克隆.结论:RNA干扰下调白血病T细胞PPP2R5C基因表达抑制细胞增殖时,在一定程度上影响TCR Vβ亚家族T细胞增殖和克隆模式变化,对正常T细胞功能可能存在一定影响.

STUDIES ON THE RELATIONSHIP BETWEEN CHROMOSOME No15 AND THE BIOLOGICAL CHARACTERISTICS OF MALIGNANT TRANSFORMED SYRIAN HAMSTER FIBROBLAST CELL LINES

During the course of malignant transformation of mammalian cells in vitro,a regular varia-tion in chromosome number and structure is usually found.In order to elucidate the rela-tionship between chromosomal changes and maligannt expression,we isolated fire clonesfrom a malignant transformed Syrian hamster fibroblast cell line and analyzed their biologicalcharacteristics,as well as chromosomal changes.A positive correlation betweetn chromosomeNo 15 monosomy and the transformed and malignant phenotype was observed.We suggestthat a suppression gene may be located on chromosome No 15,and its deletion or the lossof the chromosome may result in expression of the malignant phenotype.

Cell competition in liver carcinogenesis

Cell competition is now a well-established quality control strategy to optimize cell and tissue fitness in multicellular organisms.While pursuing this goal,it is also effective in selecting against altered/defective cells with putative(pre)-neoplastic potential,thereby edging the risk of cancer development.The flip side of the coin is that the molecular machinery driving cell competition can also be co-opted by neoplastic cell populations to expand unchecked,outside the boundaries of tissue homeostatic control.This review will focus on information that begins to emerge regarding the role of cell competition in liver physiology and pathology.Liver repopulation by normal transplanted hepatocytes is an interesting field of investigation in this regard.The biological coordinates of this process share many features suggesting that cell competition is a driving force for the clearance of endogenous damaged hepatocytes by normal donor-derived cells,as previously proposed.Intriguing analogies between liver repopulation and carcinogenesis will be briefly discussed and the potential dual role of cell competition,as a barrier or a spur to neoplastic development,will be considered.Cell competition is in essence a cooperative strategy organized at tissue level.One facet of such cooperative attitude is expressed in the elimination of altered cells which may represent a threat to the organismal community.On the other hand,the society of cells can be disrupted by the emergence of selfish clones,exploiting the molecular bar codes of cell competition,thereby paving their way to uncontrolled growth.

MALIGNANT HISTIOCYTOSIS: A STUDY ON CLINICOPATHOLOGICAL FEATURES AND CELL ORIGIN

Thirty- one autopsy cases previously diagnosed as malignant histiocytosis (MH) were studied by means of immunohistochemical staining. Antibodies detecting the formalin resistant epitopes on T- cells, B- cells and those of histiocyte/monocyte origin were used. It was shown that the malignant histiocytes reacted only to the cell markers derived from histlocyte/monocyte. and only a part of lymphocytes showed positive reaction to the T and B cell markers. It is suggested that the histiocyte/monocyte lineage is the possible origin of the malignant proliferating cells in MH. The clinicopathological features and the differentiation of MH from familial erythrophagocytic lymphohistiocytosis, virus-associated hemophagocytic syndrome and malignant lymphoma are described. The pathogenesis. the causes of death and the points for attention in the treatment of MH are also discussed.

肿瘤患者T细胞受体多样性和克隆性的高通量测序分析

目的:探讨肺癌和食管癌患者外周血T细胞受体(T cell receptor,TCR)多样性和克隆分布,以及食管癌外周血和肿瘤浸润淋巴细胞中T细胞TCR的差异情况。方法:收集自2019年4月至2021年5月郑州大学第一附属医院未行放、化疗,并排除严重感染性疾病、自身免疫性疾病以及近期使用免疫抑制剂治疗史的8例食管癌、8例肺癌患者的外周血标本,选取其中1例食管癌患者肿瘤组织标本进行TCR测序。17例健康人外周血标本作为对照组。分析肿瘤患者与健康人之间以及早期、晚期肿瘤患者之间TCR多样性指数(D50)、VJ基因使用频率以及CDR3序列克隆型频率的差异。结果:食管癌、肺癌患者外周血TCR CDR3序列的D50值[分别为(0.031±0.028)和(0.077±0.034)],均低于健康人[(0.145±0.057),P<0.000 1和P=0.005 3]。食管癌、肺癌外周血TCR CDR3最大克隆比例[分别为(10.640±7.640)%和(8.334±5.575)%],均高于健康人[(4.070±2.341)%, P=0.003 1和P=0.011 9]。且TCR CDR3序列的最大克隆比例与D50值呈负相关。食管癌和肺癌患者早期外周血T细胞TCR多样性(D50)[分别为(0.057±0.028)和(0.106±0.007)]明显高于晚期患者[分别为(0.015±0.014,P=0.028 5)和(0.059±0.030,P=0.041 7)]。随着疾病进展,肺癌和食管癌患者外周血TCR CDR3中TRBV和TRBJ基因组合使用减少,单个或多个TCR克隆型占比明显增高,存在明显的单一性。结论:肺癌和食管癌中外周血TCR谱多样性低于健康人,且存在克隆性增殖。在肿瘤发展中,TCR谱多样性降低,提示TCR谱为肿瘤的诊断、免疫评估提供了潜在的生物标志物,具有重要的临床指导意义。

Stable clonal expansion of the T-cell receptor Vβ6, Vβ17 and Vβ19 T cells in a cGVHD case using genescan analysis

Objective To investigate the distribution and clonality of the T-cell receptor (TCR) Vβ repertoire in chronic graft versus host disease (cGVHD).Methods The complementarity determining region 3 (CDR3) of the TCRβ gene with 24 variable regions was amplified in peripheral blood mononuclear cells drawn from one cGVHD patient after allogenic bone marrow transplantation (allo-BMT) 35, 39, 43 or 45 months respectively, using RT-PCR, to observe the expression of TCR Vβ repertoire T cells. The PCR products were further analyzed by genescan to evaluate clonality of T cells. Ressults Fourteen or 16 TCR Vβ subfamily T ceils were detected in each sample of cGVHD case. Oligoclonal T cells were identified in TCR Vβ 6, 16, 17, 19 and 21 subfamilies. The stable clonal T cells in all samples were identified in Vβ6, Vβ17 and Vβ21 subfamilies.Conclusion Skewing distribution and stable clonal expansion of T cells can be found in cGVHD cases and it may be related to the initiation of cGVHD.

湘云鲫2号肠道干细胞标志物LGR6分子特征及其多克隆抗体制备

【目的】克隆湘云鲫2号肠道干细胞标志物LGR6(3nLGR6)并制备多克隆抗体,为从肠道干细胞角度揭示湘云鲫2号的抗病机制提供技术支持。【方法】通过PCR扩增3nLGR6基因开放阅读框(ORF)序列,采用InterProScan、TMHMM Server 2.0、ExPASy Proteomics Server、SignalP 5.0、PSORTⅡPrediction、SoftBerry-Psite、SWISS-MODEL及MEGA 11.0等在线软件进行生物信息学分析;利用原核表达系统表达融合蛋白,以纯化的融合蛋白免疫BALB/c雌性小鼠制备多克隆抗体,基于免疫组织化学分析3nLGR6在湘云鲫2号肠组织中的分布情况。【结果】3nLGR6基因ORF序列全长2874 bp,共编码957个氨基酸残基;3nLGR6蛋白理论分子量约105.4 kD,理论等电点(p I)为5.44,在其N端有1个亮氨酸重复序列结构域,C端存在1个7次跨膜结构的GPCR结构域,且在18Gly与19Ser间存在信号肽切割位点(Sec信号肽)。3nLGR6与其他硬骨鱼类的LGR6氨基酸序列高度同源,其中与二倍体斑马鱼的相似性为92.28%。与鱼类LGR6作用密切的基因有LGR4、RNF43、RSPO2、CTNNB1、APC和CTNNA1,主要涉及Wnt信号通路和Adherens junction信号通路。以纯化得到的融合蛋白3nLGR6免疫BALB/c雌性小鼠,能成功制备出小鼠抗3nLGR6多克隆抗体,且免疫组化试验结果显示该抗体可特异性识别湘云鲫2号肠道组织中的3nLGR6。【结论】3nLGR6与其他物种的LGR6氨基酸序列高度同源,其结构和功能相对较保守。制备获得的小鼠抗3nLGR6多克隆抗体能特异性识别湘云鲫2号肠道内源性3nLGR6,为后续深入研究LGR6在硬骨鱼组织中的表达情况及揭示LGR6在肠道黏膜稳态中的作用机制提供了技术支撑。

基于p38 MAPK信号通路探讨山柰酚对牙周膜细胞活力、增殖和克隆形成的作用研究

目的 探讨山柰酚在人牙周韧带源性间充质干细胞(h PDL MSC)活力、增殖和克隆形成中的作用及对p38丝裂原活化蛋白激酶(MAPK)信号通路的调控作用。方法 体外培养hPDL MSC细胞系,分为0μmol·L^(-1)、0.01μmol·L^(-1)、0.1μmol·L^(-1)、1μmol·L^(-1)、10μmol·L^(-1)、100μmol·L^(-1)山柰酚组,SB203580组,10μmol·L^(-1)山柰酚+SB203580组。分别采用细胞计数试剂盒-8法、5-乙基-2’-脱氧尿苷法、平板克隆法、Western blotting法检测细胞活力、增殖、克隆形成及p38 MAPK通路蛋白水平。结果 药物处理48 h后,与0μmol·L^(-1)山柰酚组相比,0.1μmol·L^(-1)、1μmol·L^(-1)、10μmol·L^(-1)山柰酚组细胞活力升高(P <0.05),且细胞形状从细长变成更立方体;药物处理48 h后,与0μmol·L^(-1)山柰酚组相比,0.1μmol·L^(-1)、1μmol·L^(-1)和10μmol·L^(-1)山柰酚组EDU阳性率升高,1μmol·L^(-1)、10μmol·L^(-1)山柰酚组克隆形成数增多,10μmol·L^(-1)山柰酚组p-p38蛋白表达升高(P <0.05);与10μmol·L^(-1)山柰酚+SB203580组相比,10μmol·L^(-1)山柰酚组p-p38蛋白水平、EDU阳性率、克隆形成数较高,而SB203580组p-p38蛋白表达、细胞活力、EDU阳性率、克隆形成数较低(P <0.05)。结论 山柰酚可促进hPDL MSC细胞活力、增殖和克隆形成能力,且激活p38 MAPK通路是其分子机制之一。