Correlation of Cytotoxic Effect of Transmembrane and Secretory TNF-α to Cell Cycle

This study was aimed to examine the correlation of the cytotoxic effects induced by two types of TNF-α to cell cycle. Hoechst 33342 and PI were used to detect the morphological changes in the cell death induced by the two types of TNF-α. TdT and PI co-staining was performed to determine the phase of cell cycle of apoptotic cells. L929 cells in different phases of cell cycle were further synchronized and their sensitivity to the two types of TNF-α was observed. Our results showed that the apoptosis of HepG2 cells triggered by tm-TNF-α mainly occurred in G1 phase while in HL-60, Raji and K562 cell lines it mainly took place in S phase. The apoptosis of L929 cells induced by tm-TNF-α mainly occurred in S phase while the apoptosis induced by s-TNF-α mainly appeared in G1 phase. L929 cells were sensitive to s-TNF-α when synchronized in G1 phase (cytotoxicity 49.8%) while their sensi-tivity to tm-TNF-α was highest in S phase (45.7%) and G1/S phase (cytotoxicity 40.6%). It was concluded that tm-TNF-α-induced apoptosis of different target cells took place in different phases of cell cycle. The apoptosis of the specific cell line induced by the two types of TNF-α occurred in different phases of cell cycle. The sensitivity of the specific cell line to the two types of TNF-α was correlated with the phase of cell cycle.

The Experimental and Clinical Study on the Effect of Curcumin on Cell Cycle Proteins and Regulating Proteins of Apoptosis in Acute Myelogenous Leukemia

To investigate whether the Bcl- 2 gene family is involved in m odulating mechanism of apoptosis and change of cell cycle protein induced by curcumin in acute myeloid leukemia HL - 6 0 cell line and primary acute m yelogenous leukem ic cells,the Bcl- 2 family member Mcl- 1,Bax and Bak and cell cycle proteins including P2 7kipl,P2 1wafl,cyclin D3and p Rbp- were selected and their ex- pression detected by SABC imm uno- histochem ical stain m ethod.The attitude of sub- G1 peak in DNA histogram was determined by FCM.The TU NEL positive cell percentage was identified by term inal deoxynucleotidyl transferase (Td T ) - m ediated Biotin d U NP end labeling technique.It was found that when HL - 6 0 cells were treated with 2 5μm ol/ L curcumin for 2 4 h,the expression level of Mcl- 1was down- regulated,but that of Bax and Bak up- regulated time- dependently.There was significant difference in the expression level of Mcl- 1,Bax and Bak between the curcumin- treated groups and control group(P<0 .0 5 - 0 .0 1) .At the sam e time,curcumin had no effect on progress of cell cycle in prim aty acute m yelogenous leukemia at newly diagnosis,but could in- crease the peak of Sub- G1 (P<0 .0 5 ) ,and down- regulate the expression of Mcl- 1and up- regulate the expression of Bax and Bak with the difference being statistically significant.The expression of P2 7kipl,P2 1wafl and p Rbp- were elevated and thatof cyclin D3decreased in the presence of curcum in. These findings suggested thatthe Bcl- 2 gene fam ily indeed participated in the regulatory process of apoptosis induced by curcumin in HL - 6 0 cells and AML cells.Curcumin can induce apoptosis of primary acute myelogenous leukemic cells and disturb cell cycle progression of HL - 6 0 cells.The m echanism appeared to be m ediated by perturbing G0 / G1 phases checkpoints which associated with up- regulation of P2 7kipl,P2 1wafl and p Rbp- expression,and down- regulation of cyclin D3.

Comparison of Cell Deaths Induced by Transmembrane and Secretory TNF-α

Our previous study showed that transmembrane TNF-α （TM-TNF-α） had broader tumoricidal spectrum than secretory TNF-α （s-TNF-α）. This study examined the difference between the two kinds of TNF-α in inducing cells and the relationship between the apoptosis induced by TM-TNF-α and the cell cycle. Bioassay was employed to compare the cytotoxic effect of two kinds of TNF-α on cell lines L-929 and HepG2. TUNEL was used to detect apoptosis and the TdT and PI co-staining were used for determining the phase of apoptotic cells. Our results showed that TM-TNF-α could kill not only s-TNF-sensitive L929 cells but also s-TNF-tolerant HepG2 cells. TM-TNF-α predominantly induced apoptosis while s-TNF could induce both apoptosis and necrosis. The apoptosis of L-929 cells induced by TM-TNF-α mainly occurred in S phase and the apoptosis of HepG2 predominantly took place in G1 phase. It is concluded that the cytotoxic effects of the two TNF differ substantially. Since TM-TNF-α works locally, mainly induces apoptosis and has broader anti-tumor spectrum, it may be more effective for the treatment of tumor than s-TNF.

Biological function of miRNA-145-5p in angiotensin II induced renal inflammation

Objective:Chronic kidney disease(CKD)is a progressive disorder characterized by intricate structural and functional alterations in the kidneys,attributable to diverse causative factors.Notably,the therapeutic promise of miR-145-5p in addressing renal pathologies has been discerned.This investigation seeks to elucidate the functional role of miR-145-5p in injured kidneys by subjecting human glomerular mesangial cells(HGMCs)to stimulation with Angiotensin II(AngII).Materials and Methods:Cellular viability and the levels of inflammatory mediators were evaluated utilizing Cell Counting Kit-8(CCK-8),quantitative real-time polymerase chain reaction(qRT-PCR),and western blot methodologies,both in the presence of AngII incubation and in scenarios of miR-145p overexpression and downregulation.Furthermore,the cell cycle dynamics were elucidated through Fluorescence-activated Cell Sorting(FACS)analysis.Results:AngII incubation induced an upregulation of miR-145-5p and inflammatory factors including Intercellular Adhesion Molecule 1(ICAM-1),Interleukin 6(IL-6),Interleukin 8(IL-8),and Interleukin 1β(IL-1β).Additionally,it elevated the expression of Cyclin A2,Cyclin D1,and the G2/M cell cycle ratio.Conversely,inhibition of miR-145-5p heightened the levels of inflammatory factors and cell cycle regulators induced by AngII incubation.Reduced expression of miR-145-5p correlated with a downregulation of Interleukin 10(IL-10)expression,concurrently promoting HGMC proliferation under AngII stimulation.Moreover,ectopic miR-145-5p expression demonstrated a reduction in inflammatory factors,cell cyclin regulators,G2/M cell cycle ratio,and overall proliferation.Conclusion:MiR-145-5p exhibited inhibitory effects on the inflammatory response and proliferation induced by Angiotensin II in HGMCs,showcasing its potential as a therapeutic avenue for the treatment of kidney injury.

TUMOR NECROSIS FACTOR－α ALTERS PROTEINMETABOLISM AND CELL－CYCLE KINETICSIN MALIGNANT TUMOR

The effects of tumor necrosis factor-α(TNF) on protein metabolism and cell-cycle kinetics were investigated in malignant tumor. Sprague-Dawley rats, subcutaneously inoculated with Walker 256 carcinosarcoma,were injected intraperitoneally with recombinant human TNF at a dose of 4-75×106 U/kg for 3 consecutive days.Tumor protein metabolism and cell-cycle kinetics were analyzed. The results showed a significant decrease in tumor volume and weight in comparison with control.TNF resulted in significant decrease in tumor Protein fractional synthesis rate, Protein synthesis and fractional growth rate, but no change of tumor protein fractional degradation rate. TNF also resulted in remarkable decline in labelling index and GI phase increase of tumor cells, 6 hours after bromodeoxyuridine injection, by cytometry. The results indicated that TNF inhibits tumor growth as a result of decreases in tumor cell DNA and protein syntheses.

东北鹤虱水提取物对小鼠脾淋巴细胞体外增殖、分泌IL-2和TNF-α的作用

目的:观察东北鹤虱水提取物(LEG-I)对小鼠脾淋巴细胞体外增殖、分泌IL-2和TNF-α的作用。方法:运用噻唑兰比色(MTT)法检测LEG-I对静止状态及刀豆蛋白A(ConA)活化的淋巴细胞增殖反应的影响;用碘化丙啶染色,流式细胞术对经LEG-I作用的淋巴细胞周期进行分析;分别用MTT法和ELISA检测LEG-I刺激后静止及活化淋巴细胞分泌TNF-α和IL-2的水平。结果:LEG-I可刺激小鼠脾淋巴细胞以及被ConA活化的淋巴细胞增殖,在一定范围内具有剂量依赖性(P<0.01);相比空白对照组,LEG-I组的G0/G1期淋巴细胞比例降低,而S期和G2/M期比例增加(P<0.01);LEG-I对静止状态的淋巴细胞分泌TNF-α及IL-2无作用,而使ConA激活的淋巴细胞的分泌增加(P<0.01)。结论:LEG-I在免疫激活或免疫增强方面发挥了重要作用。

TNF与U937细胞的bcl-2基因表达及细胞周期的关系

目的:探讨肿瘤坏死因子(TNF)与白血病U937细胞的bcl—2基因表达和细胞周期的关系。方法:首先用TNF处理U937细胞,提取小片段DNA进行DNA断裂分析,继而用RT—PCR和S—P免疫组化染色法检测U937细胞的bcl—2基因表达变化,并通过流式细胞术分析TNF对U937细胞的DNA含量和细胞周期的影响。最后,用磷酸钙沉淀法将bcl—2基因转染到U937细胞,经G418筛选稳定转染子,用不同剂量TNF处理后观察转染与未转染细胞的存活率。结果:TNF可诱发U937细胞凋亡,经TNF处理的U937细胞bcl—2基因表达下降,细胞周期阻滞于G0/G1期。用bcl—2基因稳定转染的U937细胞经TNF处理,存活率明显高于未转染细胞。结论:TNF下调U937细胞bcl—2基因表达、改变细胞周期是其诱导细胞凋亡的可能作用机制。

Lanthanides-Induced Cellular Signal Transduction: Implications in Pathogenesis of Fibrosis

With the extensive mining and application of lanthanides in China and worldwide, the potential impact of lanthanides on human health is gaining increasing attentions. The recent etiological association of gadolinium-based contrast agents with nephrogenic systemic fibrosis (NSF) evoked widespread concerns regarding the safety issue of lanthanides. The elucidation of the cellular biological effects of and the signalling cascade induced lanthanides is essential for proper evaluation of their health impacts.

肿瘤坏死因子α通过激活NF-κB信号通路加快肝细胞周期进程

在慢性炎症部位有易发肿瘤的倾向,大约有20%的恶性肿瘤发生与慢性炎症相关,肝细胞癌是世界第三大癌症死亡病因,其患者多数有慢性炎症病史,当炎症慢性迁延,肝细胞癌发生率明显增加.但慢性炎症与肿瘤发生与发展的细胞和分子机制仍然不清楚.利用人肝细胞株L-02细胞,研究肿瘤坏死因子α(TNF-α)对细胞周期的影响及其机制,并探讨核因子κB(NF-κB)和ERK1/2活化对细胞周期的影响,以期能更确切地阐明炎症介质TNF-α在肝细胞癌发生发展中的作用.发现TNF-α能促进肝细胞从G0/G1期向S期转换.蛋白质印迹检测表明,TNF-α能以剂量依赖方式诱导cyclinD1表达,而对cyclinE的表达无明显影响.同时TNF-α能激活NF-κB,ERK1/2,抑制NF-κB活化降低了TNF-α诱导的cyclinD1表达,导致细胞周期阻滞于G0/G1期.抑制ERK1/2活化则对细胞周期和cyclinD1表达无显著影响.结果提示,TNF-α通过活化NF-κB信号通路,诱导cyclinD1表达,加快细胞周期进程,这可能是促进肿瘤的发生发展重要机制.针对TNF-α和NF-κB的治疗可能延长慢性炎症相关性肿瘤的潜伏期和抑制肿瘤的发展.

NF-κB对肺癌细胞生长、细胞周期及肿瘤坏死因子-α分泌的影响

目的探讨核因子(NF)-κB对肺癌细胞生长、细胞周期及分泌肿瘤坏死因子(TNF)-α的影响。方法用NF-κB抑制剂SN50处理肺癌细胞,MTT测定细胞增殖,流式细胞术测定细胞周期和细胞凋亡,酶联免疫吸附试验(ELISA)测定培养液上清中TNF-α含量,Western印迹测定细胞中细胞周期蛋白(Cyclin)B1、Cyclin D1、剪切的含半胱氨酸的天冬氨酸蛋白水解酶(酶切Caspase)-3、Bcl-2相关X蛋白(Bax)、NF-κBp65蛋白水平。结果 SN50能够抑制肺癌细胞的增殖,且随着SN50作用浓度和作用时间的增加抑制增殖作用越强。SN50组凋亡率明显高于对照组(P<0.05)。SN50组肺癌细胞G2/M比例明显高于对照组(P<0.05)。SN50组TNF-α水平明显高于对照组(P<0.05),Cyclin B1、Cyclin D1、NF-κBp65蛋白水平明显低于对照组,而酶切Caspase-3、Bax水平明显高于对照组(P<0.05)。结论抑制NF-κB信号通路可以阻碍肺癌细胞增殖,将细胞周期阻滞在G2/M期,诱导细胞凋亡,促进细胞中酶切Caspase-3、Bax表达,下调细胞中Cyclin B1、Cyclin D1蛋白水平。

4-1BB-encoding CAR causes cell death via sequestration of the ubiquitin-modifying enzyme A20

CD28 and 4-1BB costimulatory endodomains included in chimeric antigen receptor(CAR)molecules play a critical role in promoting sustained antitumor activity of CAR-T cells.However,the molecular events associated with the ectopic and constitutive display of either CD28 or 4-1BB in CAR-T cells have been only partially explored.In the current study,we demonstrated that 4-1BB incorporated within the CAR leads to cell cluster formation and cell death in the forms of both apoptosis and necroptosis in the absence of CAR tonic signaling.Mechanistic studies illustrate that 4-1BB sequesters A20 to the cell membrane in a TRAF-dependent manner causing A20 functional deficiency that in turn leads to NF-κB hyperactivity,cell aggregation via ICAM-1 overexpression,and cell death including necroptosis via RIPK1/RIPK3/MLKL pathway.Genetic modulations obtained by either overexpressing A20 or releasing A20 from 4-1BB by deleting the TRAF-binding motifs of 4-1BB rescue cell cluster formation and cell death and enhance the antitumor ability of 4-1BB-costimulated CAR-T cells.

The role of epigenetic inactivation of 14-3-3σ in human cancer

Cancer cells show characteristic alterations in DNA methylation patterns. Aberrant CpG methylation of specificpromoters results in inactivation of tumor suppressor genes and therefore plays an important role in carcinogenesis. Thep53-regulated gene 14-3-3σ undergoes frequent epigenetic silencing in several types of cancer, including carcinoma ofthe breast, prostate, and skin, suggesting that the loss of 14-3-3σ expression may be causally involved in tumor progression.Functional studies demonstrated that 14-3-3σ is involved in cell-cycle control and prevents the accumulation of chro-mosomal damage. The recent identification of novel 14-3-3σ-associated proteins by a targeted proteomics approachimplies that 14-3-3σ regulates diverse cellular processes, which may become deregulated after silencing of 14-3-3σexpression in cancer cells.

Construction of a recombinant lentivirus containing human microRNA-7-3 and its inhibitory effects on glioma proliferation

In the present study, we constructed a lentivirus, FIV-CMV-GFP-miR-7-3, containing the microRNA-7-3 gene and the green fluorescent protein gene, and used it to transfect human glioma U251 cells. Fluorescence microscopy showed that 80% of U251 cells expressed green fluorescence. Real-time reverse transcription PCR showed that microRNA-7-3 RNA expression in U251 cells was significantly increased. Proliferation was slowed in transfected U251 cells, and most cells were in the G1 phase of the cell cycle. In addition, the expression of the serine/threonine protein kinase 2 was decreased. Results suggested that transfection with a lentivirus carrying microRNA-7-3 can effectively suppress epidermal growth factor receptor pathway activity in U251 cells, arrest cell cycle transition from GI phase to S phase and inhibit glioma cell growth.

敲低金属蛋白酶组织抑制物1(TIMP-1)抑制TGF-β1诱导的MRC-5人胚肺成纤维细胞增殖和胶原蛋白合成

目的探讨敲低金属蛋白酶组织抑制物1(TIMP-1)对转化生长因子β1(TGF-β1)诱导的MRC-5人胚肺成纤维细胞生长的影响。方法将MRC-5细胞分为空白组、 TGF-β1组、转染阴性对照小干涉RNA的TGF-β1组和转染TIMP-1小干涉RNA(siTIMP-1)的TGF-β1组。噻唑蓝(MTT)法检测细胞活力,流式细胞术检测细胞周期分布, ELISA检测上清液肿瘤坏死因子α(TNF-α)含量, Western blot法检测TIMP-1、α平滑肌肌动蛋白(α-SMA)、 1型胶原蛋白(Col1)和β联蛋白(β-catenin)的蛋白水平。结果转染siTIMP-1组的MRC-5细胞TIMP-1蛋白水平明显降低。与空白组相比, TGF-β1组细胞活力明显升高、 G0/G1期细胞百分比明显降低、 S期和G2/M期细胞百分比明显升高, TNF-α含量明显升高,α-SMA、 Col1和β-catenin的蛋白水平均明显升高。与TGF-β1组相比,敲低TIMP-1水平后,细胞活力明显降低、 G0/G1期细胞百分比明显升高、 S期和G2/M期细胞百分比明显降低, TNF-α含量明显降低,α-SMA、 Col1和β-catenin蛋白表达均明显降低。转染阴性对照小干涉RNA的细胞与TGF-β1组各项指标均无明显差异。结论敲低TIMP-1水平可抑制MRC-5细胞增殖活性、减少胶原蛋白合成及TNF-α释放,可能与抑制Wnt/β-catenin信号通路有关。

Effects of galectin-3 inhibition on endometrial cell cycle and adhesion

Galectin-3(gal-3)and its ligands have been implicated in cell transformation and cancer metastasis.Gal-3 protein has been found in uterine epithelial cells adjacent to implanting blastocysts in different cell types.In order to investigate the role of gal-3 in the establishment of human endometrial receptivity,the expression of gal-3 in human endometrial cell line RL95-2 was silenced by RNA inter-ference technology using gal-3 specific small RNA.The expression of gal-3 was detected by the reverse transcriptase-polymerase chain reaction and Western blot analysis.After the suppression of gal-3,cell cycle changes and the expres-sion of integrin b1 were detected by flow cytometry.The adhesive ability of RL95-2 cells was analyzed by the adhesion test.Gal-3 siRNA transfection efficiency reached 70%–90%.The expression of gal-3 mRNA and protein in RL95-2 cells was strongly inhibited by 70%–90%after RNA interference.Inhibition of gal-3 expression decreased S-phase but increased G1 phase cells.Integrin b1 expression was down-regulated,and the adhesive ability of RL95-2 cells to fibronectin(FN)was significantly reduced.Gal-3 may be involved in the establishment of endometrial receptivity by regulating the proliferation and adhesion of endometrial cells.The influence on adhesion may be related with the integrin modulation.

Epigenetic Enabled Normal Human Cells, Lead to <i>First Cell</i>’s Unique Division System, Driving Tumorigenesis Evolution

Normal cells must become cancer-enabling before anything else occurs, according to latest literature. The goal in this mini-review is to demonstrate special tetraploidy in the enabling process. This we have shown from genomic damage, DDR (DNA Damage Response) activity with skip of mitosis leading to diploid G2 cells at the G1 border in need of chromatin repair for continued cell cycling to the special tetraploid division system. In several studies specific methylation transferase genes were activated in normal human cells in tissue fields, containing different cell growth stages of the cancerous process. Histology studies, in addition to molecular chemistry for identification of oncogenic mutational change, were a welcome change (see below). In a study on melanoma origin, DDR also showed arrested diploid cells regaining cycling from methylation transferase activity with causation of 2n melanocytes transforming to 4n melanoblasts, giving rise to epigenetic tumorigenesis enabled First Cells. Such First Cells were from Barrett’s esophagus shown to have inherited the unique division system from 4n diplochromosomal cells, first described in mouse ascites cancer cells (below). We discovered that the large nucleus prior to chromosomal division turned 90° relative to the cytoskeleton axis, and divided genome reductive to diploid, First Cells, in a perpendicular orientation to the surrounding normal cells they had originated from. This unique division system was herein shown to occur at metastasis stage, implying activity throughout the cancerous evolution. Another study showed 4-chromatid tetraploidy in development to B-cell lymphoma, and that such cancer cells also proliferated with participation of this unusual division system. Such participation has long been known from Bloom’s inherited syndrome with repair chiasmas between the four chromatids, also an in vitro observation by us. Our cytogenetic approach also revealed that they believed mitotic division in cancer cells is wrong because such cell divisions were found to be from an adaptation between amitosis and mitosis, called amitotic-mitosis. Amitosis means division without centrosomes, which has long been known from oral cancer cells, in that MOTCs (microtubule organizing center) were lacking centrioles. This observation calls for re-introduction of karyotype and cell division studies in cancer cell proliferation. It has high probability of contributing novel approaches to cancer control from screening of drugs against the amitotic-mitotic division apparatus.

The cell cycle inhibitor P21 promotes the development of pulmonary fibrosis by suppressing lung alveolar regeneration

The cell cycle inhibitor P21 has been implicated in cell senescence and plays an important role in the injury-repair process following lung injury.Pulmonary fibrosis(PF)is a fibrotic lung disorder characterized by cell senescence in lung alveolar epithelial cells.In this study,we report that P21 expression was increased in alveolar epithelial type 2 cells(AEC2 s)in a time-dependent manner following multiple bleomycin-induced PF.Repeated injury of AEC2 s resulted in telomere shortening and triggered P21-dependent cell senescence.AEC2 s with elevated expression of P21 lost their self-renewal and differentiation abilities.In particular,elevated P21 not only induced cell cycle arrest in AEC2 s but also bound to P300 andβ-catenin and inhibited AEC2 differentiation by disturbing the P300-β-catenin interaction.Meanwhile,senescent AEC2 s triggered myofibroblast activation by releasing profibrotic cytokines.Knockdown of P21 restored AEC2-mediated lung alveolar regeneration in mice with chronic PF.The results of our study reveal a mechanism of P21-mediated lung regeneration failure during PF development,which suggests a potential strategy for the treatment of fibrotic lung diseases.

LncRNA ZFPM2-AS1 promotes phyllodes tumor progression by binding to CDC42 and inhibiting STAT1 activation

Breast phyllodes tumor(PT)is a rare fibroepithelial neoplasm with potential malignant behavior.Long non-coding RNAs(lncRNAs)play multifaceted roles in various cancers,but their involvement in breast PT remains largely unexplored.In this study,microarray was leveraged for the first time to investigate the role of lncRNA in PT.We identified lncRNA ZFPM2-AS1 was significantly upregulated in malignant PT,and its overexpression endowed PT with high tumor grade and adverse prognosis.Furthermore,we elucidated that ZFPM2-AS1 promotes the proliferation,migration,and invasion of malignant PT in vitro.Targeting ZFPM2-AS1 through nanomaterial-mediated siRNA delivery in patient-derived xenograft(PDX)model could effectively inhibit tumor progression in vivo.Mechanistically,our findings showed that ZFPM2-AS1 is competitively bound to CDC42,inhibiting ACK1 and STAT1 activation,thereby launching the transcription of TNFRSF19.In conclusion,our study provides evidence that ZFPM2-AS1 plays a pivotal role in the pathogenesis of breast PT,and suggests that ZFPM2-AS1 could serve as a prognostic indicator for patients with PT as well as a promising novel therapeutic target.

E2F4 regulates the cell cycle and DNA replication in the silkworm,Bombyx mori

The E2F family of transcription factors is crucial for cell cycle progression and cell fate decisions.Although E2Fs have been widely studied in mammals,there have been few studies performed in insects.Here,we determined the function of E2F4 in the silkworm,Bombyx mori.We demonstrate that E2F proteins are highly conserved among species from lower animals to higher mammals.Overexpression of the BmE2F4 gene led to cell cycle arrest in the G1 phase,whereas interfering with the BmE2F4 mRNA led to accumulation of cells in the S phase.These results indicate that BmE2F4 is important in cell cycle regulation.We also demonstrate that the BmE2F4 gene is involved in DNA replication of BmN-SWU1 cells and DNA synthesis in the silk gland.Furthermore,we identified a protein called Bm14-3-3ζthat can interact with BmE2F4 and allow it to localize in the nucleus.Overexpression of the Bm14-3-3ζgene led to cell cycle arrest in the G1 phase,while knocking down the gene increased the proportion of cells in S phase.These findings provide important insights into the function of E2F transcription factors and increase our understanding of their involvement in cell cycle regulation.

参红补血颗粒对失血性血虚模型小鼠骨髓细胞周期及肾脏TNF-α表达的影响

目的:观察参红补血颗粒对失血性血虚模型小鼠骨髓细胞周期及肿瘤坏死因子-α(TNF-α)表达的影响。方法:尾动脉放血法复制失血性血虚证动物模型。60只小鼠随机分为空白对照组,模型组,阳性对照组,参红补血颗粒高、中、低剂量组,每组10只,连续给药14d。血液分析仪检测红细胞(RBC)、血红蛋白(HGB)、红细胞比容(HCT)、白细胞(WBC)及血小板(PLT)水平,流式细胞术检测骨髓细胞周期变化,Western Blot检测肾脏TNF-α蛋白表达。结果:与模型组比较,阳性对照组及参红补血颗粒高、中、低剂量组RBC、HGB、HCT、WBC及PLT水平均有不同程度的提高(P<0.05);阳性对照组、参红补血颗粒高、中剂量组处于G2/M期的骨髓细胞数量显著性增多(P<0.01),S期细胞显著性减少(P<0.01);阳性对照组及参红补血颗粒高、中剂量组肾脏TNF-α表达水平显著性降低(P<0.05)。结论:参红补血颗粒对失血性血虚模型小鼠的造血及免疫功能有明显的改善作用,其作用机制可能与促进骨髓细胞增殖及降低肾脏TNF-α蛋白表达水平有关。