Formulation And Optimization Of Terbinafine Hcl Solid Lipid Nanoparticles For Topical Antifungal Activity

Objective: Solid lipid nanoparticles (SLNs) are at the forefront of the rapidly developing field of nanotechnology with several potential applications in drug delivery and research. The aim of this study was to develop and characterize SLNs formulae of Terbinafine HCl (TFH) for topical drug delivery applications. Methods: SLNs were prepared using the solvent injection technique. Glyceryl Monostearate (GMS) served as the lipid base. Three stabilizers; Tween 80, Cremophor RH40, and Poloxamer 188, were used. The effect of stabilizer type and concentration, as well as the lipid concentration, were studied, factorial design of 32*21was applied. The prepared SLNs were characterized regarding their particle size, zeta potential, polydispersity index (PDI), entrapment efficiency percent (EE %), and physicochemical stability. The selected formulae were subjected to further investigations such as morphological studies, in vitro release studies, and Infrared (IR) spectroscopy. They were compared with the marketed cream Lamifen® in term of their antifungal activity against Candida albicans. Results: Lipid concentration, together with the type and concentration of stabilizer, appeared to be the main cornerstones which affect the formation of SLNs. Smaller particle size was observed when increasing the stabilizer concentration and decreasing the lipid concentration. Higher EE% was observed when increasing both the stabilizer and the lipid concentrations. Formulae (F6, F12 andF19) were selected as the most suitable SLNs with optimum particle size of 480.2±18.89, 458.6±12.45 and 246.7±10.5 nm, respectively as well as the highest EE% of 87.13±0.19, 93.69±0.7 and 95.06±0.25, respectively. In vitro microbiological screening of their antifungal activity showed significantly larger zones of inhibition of diameters 25.9±0.25, 25±0.35 and 24.67±0.36 mm, respectively in comparison with the marketed Lamifen® cream which showed a zone of 11.2±0.44 mm diameter. Conclusion: Applying SLNs containing TFH as topical antifungal preparations may be considered as a very promising option as they show good physicochemical characterization with high antifungal activity, which delineates them as a promising dosage form for topical antifungal treatment.

Curcumin Elevates T(FH) Cells and Germinal Center B Cell Response for Antibody Production in Mice

Curcumin is a natural product extracted from Curcuma longa. It has been reported as a potent antioxidant and anti-inflammatory compound. Previous studies have demonstrated that curcumin suppresses pro-inflammatory cytokine production via inhibition of NF-κB in macrophages. However, its role in adaptive immune cells such as T cells, in vivo, has not clearly been elucidated. Here, we examined the effects of curcumin in T follicular helper (T(FH)) cells and on Ab production during NP-ovalbumin immunization in mice. The results revealed that curcumin administered daily significantly increased CXCR5⁺B-cell lymphoma 6⁺ T(FH) cells and CD95⁺GL-7⁺ germinal center (GC) B cells in draining lymph nodes. In addition, curcumin treatment in mice induced total Ab production as well as high affinity IgG1 and IgG2b Ab production. Collectively, these results suggest that curcumin has positive regulatory roles in T(FH) cell functions and GC responses. Thus, this could be an advantageous supplement to enhance humoral immunity against infectious diseases and cancer.

Characteristics and clinical significance of cellular immunity indices of tfh cells in HIV-1 infected patients / 解放军医学杂志

[Abstract] Objective To study the correlation between follicular helper T (TFH) cells with the cellular immunity indices and the percentage of follicular cytotoxic T (TFC) cells in peripheral blood of human immunodeficiency virus-1 (HIV-1) infected patients and explore the relationship between TFH cells and the acquired immunodeficiency symdrome (AIDS) disease progression. Methods Seventeen health controls and 65 HIV-1 infected individuals were enrolled in this study, and the HIV-1 infected patients included 48 untreated HIV-1 positive patients and 17 treated with highly active antiretroviral therapy, the 48 HIV-1 infected patients consisted of 9 patients with CD4+ T cells ≤200/μl, 19 with 200/μl 350/μl. The expression of CXCR5 on CD4+ T cells was detected by flow cytometry and its correlation with CD4/CD8 ratio and HIV-1 RNA viral load. We also analyzed the correlation between the percentage and absolute number of TFH cells and TFC cells. Results Compared with healthy controls, both the absolute cell numbers [(60.23±27.62)/μl vs. (207.53±51.50)/μl, U=0, P<0.0001] and frequency [(18.83±4.55)% vs. (22.06±5.35)%, U=242.00, P=0.0124] of TFH cells were significantly reduced in untreated HIV-1 infected patients; The absolute number of TFH cells in the antiretroviral therapy group was significantly higher than that in untreated HIV-1 infected patients [(135.05±60.77)/μl vs. (60.23±27.62)/μl, U=108.00, P=0.0082]. The frequency of TFH cells in untreated HIV-1 infected patients were negatively correlated with CD4/CD8 ratio (r=–0.338, P=0.019), positively correlated with viral load (r=0.392, P=0.006). Both the absolute cell numbers (r=0.408, P=0.004), and frequency (r=0.363, P=0.011) of TFH cells were positively correlated with the frequency of TFC cells. The percentage of TFH cells decreased with the decrease of CD4+ T cells and the number of TFH cells increased with the decrease of CD4+ T cells in untreated HIV-1 infected patients. Conclusions TFH cells may be associated with active replication of the HIV-1 and disease progression. The decrease of TFH absolute cell numbers in patients with HIV-1 infection may be related to the cytotoxicity of TFC cells.

Flow cytometry analysis of circulating and splenic follicular helper T ( Tfh) cells in a mouse model of allergic rhinitis / 中华微生物学和免疫学杂志

Objective To investigate changes in the percentages of circulating and splenic follicular helper T ( Tfh) cells and the significance of Tfh cells in a mouse model of allergic rhinitis .Meth-ods BALB/c mice were sensitized by injection of OVA and aluminum hydroxide to establish the model of allergic rhinitis .Flow cytometry was performed to measure the percentages of circulating and splenic CD 4+CXCR5+after staining Tfh cells with anti-mouse CD4-FITC and anti-mouse CXCR5-PE.Concentrations of OVE-specific IgE ( OVA-sIgE ) in serum samples were measured by enzyme-linked immunosorbent assay ( ELISA ) .Student′s t-test and Pearson ′s correlation analysis were used for statistical analysis .Results Enhanced infiltration of eosinophils in submucosa was observed in the experimental group .After sensitizing the mice with OVA and aluminum hydroxide , the concentration of OVA-sIgE increased from 43 .47 pg/ml [(43.47±2.58) pg/ml] to 50.44 pg/ml [(50.44±1.40) pg/ml] (P=0.029); the percentage of circu-lating CD4+CXCR5+was up-regulated from 6.25％ [(6.25±1.19)％] to 13.94％ [(13.94±2.77)％] (P=0.026); the percentage of splenic CD4+CXCR5+was down-regulated from 18.04％ [(18.04 ± 4.97)％] to 7.26％[(7.26±0.96)％] (P=0.019).The concentration of OVA-specific IgE was posi-tively correlated with the percentage of circulating Tfh cells (r=0.954, P=0.002), but negatively correla-ted with the percentage of splenic Tfh cells (r=-0.801, P=0.028).Moreover, a negative correlation was found between circulating and splenic Tfh cells (r=-0.787, P=0.032).Conclusion Tfh cells might be involved in the immune responses against allergic rhinitis in mice .

Changes of peripheral follicular helper T cell subsets in HIV infected patients / 中华实验和临床病毒学杂志

Objective@#To investigate the changes of the frequencies of follicular helper T (Tfh) cell subsets in peripheral blood of HIV-1 infected patients and its relevance to the disease severity.@*Methods@#Twenty untreated HIV+ patients, 20 HAART-suppressed patients and 15 health controllers are enrolled in this study. The frequencies of Tfh and its subsets were examined by flow cytometry.@*Results@#The frequency of Tfh17 subset in peripheral blood decreased significantly in untreated HIV-1+ patients as compared with those of healthy controls and HAART-suppressed patients (P<0.05). In addition, the frequency of this subset were positively correlated with CD4+ T cell counts in both untreated and treated HIV+ patients, and were inversely correlated with viral load and plasma-immunoglobulin in untreated HIV+ patients. The frequency of Tfh2 subset, however, were inversely correlated with CD4+ T cell counts and positively correlated with viral load and plasma-immunoglobulin in untreated HIV+ patients. The frequency of Tfh1 subset in peripheral blood increased significantly in untreated and treated HIV-1+ patients as compared with those of healthy controls (P<0.05) and were inversely correlated with viral load and plasma-immunoglobulin in untreated HIV+ patients and positively correlated with CD4+ T cell counts.@*Conclusions@#The abnormal distribution of Tfh subests may play a role in the progression of HIV infection and its characteristic immune activation.

Enforced Expression of CXCR5 Drives T Follicular Regulatory-Like Features in Foxp3⁺ T Cells

CXCR5⁺ T follicular helper (Tfh) cells are associated with aberrant autoantibody production in patients with antibody-mediated autoimmune diseases including lupus. Follicular regulatory T (Tfr) cells expressing CXCR5 and Bcl6 have been recently identified as a specialized subset of Foxp3+ regulatory T (Treg) cells that control germinal center reactions. In this study, we show that retroviral transduction of CXCR5 gene in Foxp3⁺ Treg cells induced a stable expression of functional CXCR5 on their surface. The Cxcr5-transduced Treg cells maintained the expression of Treg cell signature genes and the suppressive activity. The expression of CXCR5 as well as Foxp3 in the transduced Treg cells appeared to be stable in vivo in an adoptive transfer experiment. Moreover, Cxcr5-transduced Treg cells preferentially migrated toward the CXCL13 gradient, leading to an effective suppression of antibody production from B cells stimulated with Tfh cells. Therefore, our results demonstrate that enforced expression of CXCR5 onto Treg cells efficiently induces Tfr cell-like properties, which might be a promising cellular therapeutic approach for the treatment of antibody-mediated autoimmune diseases.

Action mechanism of total flavonoids of Hippophae rhamnoides in treatment of myocardial ischemia based on network pharmacology / 中国中药杂志

In this study, a network pharmacological screening method was adopted to further study the active ingredients and action mechanism of total flavonoids of Hippophae rhamnoides(TFH) for the treatment of myocardial ischemia. Firstly TCMSP database and PubChem database were searched, and then the data were combined with oral bioavailability and drug analysis to screen flavonoids of H.rhamnoides compounds. Then predictive analysis was conducted for the 7 screened compounds by ChemMapper server.The obtained potential targets were imported into MAS 3.0. Database, and KEGG database was also used for targets analysis and pathway analysis. Finally Cystoscope 3.3.0 software was used to draw "compounds-targets-pathway" network diagram. Virtual experiments predicted 68 potential targets and 60 signaling pathways, and 31 targets and 23 pathways of them were directly or indirectly associated with myocardial ischemia. The results showed that TFH played a synergistic rolemainly through the regulation of calcium signaling pathway, VEGF signaling pathway and gap junction signaling pathway, which was consistent with literature reports. These results indicated that it can enhance heart function, protect vascular endothelial cells, and fight against myocardial ischemia probably by regulating platelet aggregation, lipid metabolism, inflammation and other processes.

Changes of circulating Tfr and Tfh cells in children with myasthenia gravis / 中华微生物学和免疫学杂志

Objective To investigate the changes of follicular regulatory T cells ( Tfr cells) and follicular T helper cells ( Tfh cells) in peripheral blood of children with myasthenia gravis ( MG) . Methods We recruited 28 MG patients and 20 healthy subjects in this study. The percentages of Tfh and Tfr cells in peripheral blood samples were measured by flow cytometry. Real-time PCR was performed to detect the ex-pression of transcription factors and regulatory factors of Bcl-6, c-MAF, Blimp-1 and PD-1 at mRNA level. ELISA was used to detect the levels of IL-2, IL-6, IL-10 and IL-21 in plasma samples and the titers of Ach-Rab and PsMab. Results Compared with the healthy subjects, the MG patients showed higher percentages of Tfh cells and lower percentages of Tfr cells before receiving treatment. The expression of Bcl-6 and c-MAF on CD4+T lymphocytes cells at transcriptional level were significantly enhanced, while the expression of Blimp-1 on CD4+T cells and the expression of PD-1 on Treg cells at transcriptional level were inhibited in the MG patients in comparison with those in healthy subjects. Moreover, decreased levels of IL-2 and increased levels of IL-21 were found in plasma samples collected from the MG patients. Conclusion The decreased percentages of Tfr cells and increased percentages of Tfh cells in patients with MG resulted in abnormal ratios of Tfr/Tfh cells, which might be involved in the immunological pathogenesis of MG. Several changes in the patients with MG might be responsible for the imbalanced ratio of Tfr/Tfh cells, which included changes of IL-2 and IL-21 in microenvironment, enhanced expression of Bcl-6 and c-MAF at mRNA level and inhibited expression of Blimp-1 at mRNA level on CD4+T cells as well as over-expression of PD-1 at mRNA level on Treg cells.

Mechanism of T follicular cells in experimental autoimmune encephalomyelitis / 中国免疫学杂志

Objective: To evaluate the mechanism of T follicular helper cells ( Tfh ) in experimental autoimmune encephalomyelitis (EAE) via in vivo experiments. Methods:C57BL/6 mice were randomly divided into four groups,CFA group,EAE group,anti-ICOSL group and control group. Lymphocytes of different time points isolated from draining lymph nodes and spleen were stained for T follicular helper cells surface marker and T cells activation surface marker and analyzed by FACS. Observed parameters include inflammatory infiltration,demyelination in spinal cord and germinal center in spleen. ELISA was used to measure the level of antigen specific antibodies. Results: Mice in anti-ICOSL treated group developed mild disease was with lower clinical scores when compared with the EAE group. HE staining results turned out with alleviated inflammation and Luxol Fast Blue staining( LFB) showed no demyelization in anti-ICOSL treated mice compared with non-treated EAE models. Flow cytometry results revealed that percentages of T follicular helper cells decreased though the whole activated degree T cells was not influenced in anti-ICOSL treated group. Fewer ger minal center was found in both anti-ICOSL group and CFA group with reduced secretion of MOG-specific Ab. Conclusion:T follicular helper cells supported the development of cognate B cells,promoted the formation of germinal center,facilitate pathogenic MOG-specific Ab secretion,thus enhance EAE.

Blockade of STAT3 in T Cells Inhibits Germinal Center Reactions against Intranasal Allergens

Understanding the developmental mechanisms of humoral immunity against intranasal antigens is essential for the development of therapeutic approaches against air-borne pathogens as well as allergen-induced pulmonary inflammation. Follicular helper T (Tfh) cells expressing CXCR5 are required for humoral immunity by providing IL-21 and ICOS costimulation to activated B cells. However, the regulation of Tfh cell responses against intranasal antigens remains unclear. Here, we found that the generation of Tfh cells and germinal center B cells in the bronchial lymph node against intranasal proteinase antigens was independent of TGF-β. In contrast, administration of STAT3 inhibitor STA-21 suppressed the generation of Tfh cells and germinal center B cells. Compared with wild-type OT-II T cells, STAT3-deficient OT-II T cells transferred into recipients lacking T cells not only showed significantly reduced frequency Tfh cells, but also induced diminished IgG as well as IgE specific for the intranasal antigens. Co-transfer study of wild-type OT-II and STAT3-deficient OT-II T cells revealed that the latter failed to differentiate into Tfh cells. These findings demonstrate that T cell-intrinsic STAT3 is required for the generation of Tfh cells to intranasal antigens and that targeting STAT3 might be an effective approach to ameliorate antibody-mediated pathology in the lung.

Follicular Helper T (Tfh) Cells in Autoimmune Diseases and Allograft Rejection

Production of high affinity antibodies for antigens is a critical component for the immune system to fight off infectious pathogens. However, it could be detrimental to our body when the antigens that B cells recognize are of self-origin. Follicular helper T, or Tfh, cells are required for the generation of germinal center reactions, where high affinity antibody-producing B cells and memory B cells predominantly develop. As such, Tfh cells are considered as targets to prevent B cells from producing high affinity antibodies against self-antigens, when high affinity autoantibodies are responsible for immunopathologies in autoimmune disorders. This review article provides an overview of current understanding of Tfh cells and discusses it in the context of animal models of autoimmune diseases and allograft rejections for generation of novel therapeutic interventions.

Follicular helper T cell in immunity and autoimmunity

The traditional concept that effector T helper (Th) responses are mediated by Th1/Th2 cell subtypes has been broadened by the recent demonstration of two new effector T helper cells, the IL-17 producing cells (Th17) and the follicular helper T cells (Tfh). These new subsets have many features in common, such as the ability to produce IL-21 and to express the IL-23 receptor (IL23R), the inducible co-stimulatory molecule ICOS, and the transcription factor c-Maf, all of them essential for expansion and establishment of the final pool of both subsets. Tfh cells differ from Th17 by their ability to home to B cell areas in secondary lymphoid tissue through interactions mediated by the chemokine receptor CXCR5 and its ligand CXCL13. These CXCR5+ CD4+ T cells are considered an effector T cell type specialized in B cell help, with a transcriptional profile distinct from Th1 and Th2 cells. The role of Tfh cells and its primary product, IL-21, on B-cell activation and differentiation is essential for humoral immunity against infectious agents. However, when deregulated, Tfh cells could represent an important mechanism contributing to exacerbated humoral response and autoantibody production in autoimmune diseases. This review highlights the importance of Tfh cells by focusing on their biology and differentiation processes in the context of normal immune response to infectious microorganisms and their role in the pathogenesis of autoimmune diseases.

Phenotypic Tfh development promoted by CXCR5-controlled re-localization and IL-6 from radiation-resistant cells

How follicular T-helper (Tfh) cells develop is incompletely understood. We find that, upon antigen exposure in vivo, both naïve and antigen-experienced T cells sequentially upregulate CXCR5 and Bcl6 within the first 24 h, relocate to the T-B border, and give rise to phenotypic Bcl6(+)CXCR5(+) Tfh cells before the first cell division. CXCR5 upregulation is more dependent on ICOS costimulation than that of Bcl6, and early Bcl6 induction requires T-cell expression of CXCR5 and, presumably, relocation toward the follicle. This early and rapid upregulation of CXCR5 and Bcl6 depends on IL-6 produced by radiation-resistant cells. These results suggest that a Bcl6(hi)CXCR5(hi) phenotype does not automatically define a Tfh lineage but might reflect a state of antigen exposure and non-commitment to terminal effector fates and that niches in the T-B border and/or the follicle are important for optimal Bcl6 induction and maintenance.

Function and correlation of follicular helper T cells and B cells in alcohol abuse non-traumatic osteonecrosis of femoral head / 中国免疫学杂志

Objective:To examine the roles of follicular helper T(Tfh)cells,serum IL-21 and B cells in the pathogenesis of alcohol abuse non-traumatic osteonecrosis of the femoral head ( NONFH ) .Flow cytometry was used to measure the frequencies of peripheral blood inducible Tfh cells and B cells in alcohol abuse NONFH patients and healthy controls.The disease progression and the extent of femoral head collapse,the serum IL-21 were quantified.Methods: A significantly higher percentages of CD19+B cells(t=3.765,P=0.0005),CD86+CD19+B cells(t=5.506,P<0.0001),and CD95+CD19+B cells(t=4.152,P=0.0002) in patients than those in controls was found.The percentages of CD86+CD19+B cells were positively associated with the index of femoral head collapse in alcohol abuse NONFH(P<0.0001,r=0.536).Results: The frequencies of Tfh cells (t=7.611,P<0.0001),and IL-21+Tfh cells (t=5.281,P<0.0001) were higher than those in controls;The frequencies of Tfh cells were positively associated with the percentages of CD19+B cells(P=0.0002,r=0.455),IL-21+Tfh cells were positively associated with the percentages of CD86+CD19+B cells(P=0.0002,r=0.447).Conclusion: Tfh cells and B cells may participate in the pathogenesis of alcohol abuse NONFH,and increased CD86+CD19+B cells may be associated with the extent of femoral head collapse,the interaction of Tfh cells and B cells may have an im-portant role in pathogenesis of alcohol abuse NONFH.

Expression and Clinical Significance of Bcl-6 mRNA in the T Follicular Helper(Tfh)Cells of HIV/AIDS Patients / 现代检验医学杂志

Objective To explore the expression of Bcl-6 mRNA in the Tfh cells of HIV/AIDS patients and the relationship between Bcl-6 mRNA and the progression of HIV/AIDS.Methods This experiment chose 60 patients who were confirmed by HIV antibody test positive,during May 2014 to November from AIDS Research Institute in the First Affiliated Hospi-tal,Henan College of Traditional Chinese Medicine.According to the amount of CD4+T cells,the patients with HIV/AIDS were divided into A(CD4500 cells/μl)three groups.The Tfh cells expressed CD4+ CXCR5 + ICOS+ and lymphocyte subpopulation including T,B and NK cells were detected by flow cytometry (FCM).The expression of Bcl-6 mRNA levels were detected by reverse transcription-polymerase chain reaction (RT-PCR).The association between Tfh cells and Bcl-6 mRNA,B cells were analyzed.Results The expression of Bcl-6 mRNA from patients of A,B and C three groups was increased than in healthy individuals (2.94±0.91,2.27±0.62,2.15± 0.351,P <0.05).The percent of Tfh cells from group A patients was higher than group C patients (5.88±3.01 vs 1.26± 0.87,P =0.032)and also the healthy control group (5.88±3.01 vs 0.78±0.42,P =0.004).The percentage of NK cells and B cells in HIV/AIDS patients was no statistically significant.Correlation analysis showed that there was positive corre-lation between the proportion of Tfh cells and Bcl-6 mRNA (r=0.799,P =0.000).And Tfh cells were observed no relation-ship with B cells (r=-0.083,P =0.657).Conclusion The changes of Bcl-6 mRNA and Tfh cells may be related to the progression of HIV/AIDS.

Characteristics of tfh cells in the peripheral blood in recipients of liver allograft: A pilot study / 解放军医学杂志

Objective To determine the expression characteristics of peripheral blood follicular helper T cells (Tfh) and their molecules during pre- and post-transplantation period in liver transplant patients to explore the role of Tfh in alloreactive response after liver transplantation. Methods Twenty liver transplant (LT) patients and 20 healthy individuals as control were enrolled in this study. Thirteen LT patients with stable liver function were included for cross-sectional study. Another seven LT patients with complete clinical data were included for follow-up study. The frequencies of Tfh cells were examined by flow cytometry. For the seven patients, Tfh cells and level of alanine aminotransferase (ALT) were monitored dynamically for one month after LT. Results The frequencies of CD4+CXCR5+Tfh and CD4+CXCR5+PD-1+ cells in peripheral blood increased significantly after liver transplantation as compared with those before liver transplantation (P<0.05). In addition, the frequencies of CD4+CXCR5+ICOS+ cells showed a rising trend, but no statistical difference. In early follow-up period it was found that the frequencies of CD4+CXCR5+ Tfh obviously increased after liver transplantation in patients with stable liver function, and reached the peak value after one week and then decreased. The AL level also decreased when reaching peak 10 days after surgery. Interestingly, the frequencies of CD4+CXCR5+ Tfh in one patient with acute rejection after liver transplantation showed the same trend with ALT levels changes. Furthermore, the frequencies of CD4+CXCR5+Tfh, CD4+CXCR5+PD-1+ and CD4+CXCR5+ICOS+ cells in stable liver function group were higher than those in the healthy control group (P<0.05). Conclusion The CD4+CXCR5+Tfh cells in peripheral blood will be upregulated in liver transplant patients, which indicates that the Tfh maybe involved in liver alloreactive response.

EFFECTS OF TOTAL FLAVONES OF HIPPOPHAE RHAM-NOIDES L (TFH) ON IMMUNOLOGICAL FUNCTION IN MICE / 中国药理学通报

TFH was isolated from the fruits of Hippophae Rhamnoides L. A study was made cf the effects of TFH on the immune reaction in animals. TFH ( 5.0mg/kg? d-1 sc?6d)remarkably enhanced the pha-gocytic activity of peritoneal exudate macrophages. The content of lysozyme in mice ( TFH 2mg/kg?d-1 ip?7d) & serum complement in guinea pig also were remarkably elevated. TFH caused significant increases of quantitative hemolysin of SRBC ( QHS ) in normal mice. At the dose of 2mg/kg?d-1 ip?8d TFH increased the production of hemolysin serum and agglutinin in normal mice as well as in immunodepressed mice induced by cyclophosphamids. TFH ( 6.25-50mg/L) markedly enhanced Con A-induced lymphocyte proliferation of mouse spleen cells in vitro. These results suggested that TFH had certain immunopotentiation in animals.

THE EFFECT OF TOTAL FLAVONES OF HIPPOPHAE RHAMNOIDES L. ON BEATING AND ELECTRICAL ACTIVITY OF CULTURED CARDIAC CELL / 西安交通大学学报(医学版)

In this paper, we studied the effect of total flavones of Hippophae rhamnoides L (TFH) on beating and transmembrane potential of cultured sucking rat cardiac cell. The results indicated that cellular beating rate and beating amplitude were significantly reduced, the action potential duration of 50% and 90% repolarization was sbortened, the slope of phase 4 depolarization was decreased, and other parameters of the action potential showed no significant change. In addition, we also observed that TFH could make the spontaneous abnormal beating of cell change into regular beating. These findings suggested that the mechanism of TFH on cultured cardiac cell might have some relation to antagonizing inward flow of Ca~(2+) and aocelerating the move of k' outward.