Study on the Viability Inhibitory Effects and Mechanism of Dictamnine on Mice Spleen Lymphocyte in vitro / 中国药房

OBJECTIVE： To study the inhibitory effect of dictamnine on the viability of mice spleen lymphocyte in vitro and explore its potential mechanism. METHODS： The primary spleen lymphocytes of mice were isolated and cultured. The cells were treated with 0 （blank control）， 50， 100， 150 μmol/L dictamnine for 24 h. MTT assay was used to determine the cell viability； Lactic dehydrogenase （LDH） assay was used to determine release rate of LDH. Early apoptosis rate was detected by flow cytometry. Necrosis rate was detected by Hoechst 33342 and PI double staining； Western blot assay was used to detect the protein expressions of Caspase 3 and Cleaved-Caspase 3 in cells. Comet assay was used to detect DNA damage in cells （reflected in the proportion of DNA tail area）. RESULTS： Compared with blank control， 100， 150 μmol/L dictamnine could significantly inhibit the viability of lymphocytes （P＜0.01）. 150 μmol/L dictamnine could significantly increase the release of LDH （P＜0.05）， and release rate reached 79.37％. 50， 100， 150 μmol/L dictamnine could improve the early apoptotic rate of lymphocyte， but there was no statistical significance （P＞0.05）. 150 μmol/L dictamnine could significantly increase the necrosis rate （P＜0.05）， and necrosis rate reached 78.64％. 50， 100， 150 μmol/L dictamnine could increase the protein expression of Caspase 3， but there was no statistical significance （P＞0.05）， while 50， 100 μmol/L dictamnine could improve the protein expression of Cleaved-Caspase 3 significantly （P＜0.05）. DNA damage was induced in a dose-dependent manner by dictamnine， in which 100 and 150 μmol/L dictamnine could significantly increase DNA tail area （P＜0.01）. CONCLUSIONS： Dictamnine can inhibit spleen lymphocyte viability， and the mechanism may be related to inducing spleen lymphocyte necrosis and DNA damage.

Effects of 1,25-dihydroxy vitamin D3 on immune function of RBC and spleen in ulcerative colitis mice / 重庆医学

Objective To investigate the effects of 1,25-dihydroxy vitamin D3 [1,25 (OH)2D3] on immune mechanism of RBC and spleen in colitis model mice.Methods Thirty mice were randomly grouped as follows:blank control group,model group and 1,25(OH)2D3 group.The animal models were established.Then the general condition and colon pathological changes in mice were observed,the changes of the RBC immune compound(RCIC) and RBC surface C3b receptor(RC3bR) wreath were detected by the yeast wreath method,the weight and length of spleen were measured,the positive rates of spleen immune cells were detected by flow cytometry.Results Compared with the model group,RBC-C3bR in the 1,25(OH)2D3 group and blank control group was significantly increased and RBC-ICR was significantly decreased (P＜0.01).Compared with the blank control group,the positive rates of CD3,CD4,CD8 and CD45R in spleen mononuclear cell were significantly increased (P＜0.01);after the 1,25 (OH) 2D3 intervention,the positive rates of CD3,CD4,CD8 and CD56R in spleen mononuclear cells were significantly decreased compared with the model group(P＜0.01).Conclusion 1,25(OH)2D3 has the immune regulatory effect on RBC and peripheral spleen lymphocytes in chronic colitis mice.

Effects of 1,25-dihydroxy vitamin D3 on immune function of RBC and spleen in ulcerative colitis mice / 重庆医学

Objective To investigate the effects of 1,25-dihydroxy vitamin D3 [1,25 (OH)2D3] on immune mechanism of RBC and spleen in colitis model mice.Methods Thirty mice were randomly grouped as follows:blank control group,model group and 1,25(OH)2D3 group.The animal models were established.Then the general condition and colon pathological changes in mice were observed,the changes of the RBC immune compound(RCIC) and RBC surface C3b receptor(RC3bR) wreath were detected by the yeast wreath method,the weight and length of spleen were measured,the positive rates of spleen immune cells were detected by flow cytometry.Results Compared with the model group,RBC-C3bR in the 1,25(OH)2D3 group and blank control group was significantly increased and RBC-ICR was significantly decreased (P＜0.01).Compared with the blank control group,the positive rates of CD3,CD4,CD8 and CD45R in spleen mononuclear cell were significantly increased (P＜0.01);after the 1,25 (OH) 2D3 intervention,the positive rates of CD3,CD4,CD8 and CD56R in spleen mononuclear cells were significantly decreased compared with the model group(P＜0.01).Conclusion 1,25(OH)2D3 has the immune regulatory effect on RBC and peripheral spleen lymphocytes in chronic colitis mice.

Effects of Drug-containing Serum of Ficus Hirta on Oxidative Damage of Spleen Lymphocyte in Aged Mice / 中国中医药信息杂志

Objective To study the effects of drug-containing serum of Ficus Hirta on oxidative damage of spleen lymphocyte due to aging in aged mice; To discuss its mechanism of action.Methods Forty aged mice were randomly divided into control group and high-, medium- and low-dose Ficus Hirta groups. Control group was given 0.9% sodium chloride solution for gavage, while high-, medium- and low-dose Ficus Hirta groups were given 6.6, 4.4, and 2.2 g/kg aqueous extract of Ficus Hirta for gavage. The spleen index was observed for optimum dose in aged mice. The optimum time and dilution of drug-containing serum of Ficus Hirta were confirmed by MTT method in lymphocyte proliferation test. The positive rate of senescent cells, the activity of T-SOD and the contents of MDA and ROS were determined in cellular antioxidant experiment after treated by optimal drug-containing serum for 48 h. Results Compared with the control group, the spleen index was significantly improved in high-, medium- and low-dose Ficus Hirta groups (P<0.05,P<0.01). 20% drug-containing serum of Ficus Hirta cultivated for 48 h had the best effects on lymphocyte proliferation in aged mice. 20% drug-containing serum of Ficus Hirta could significantly decrease the positive rate of senescent cells (P<0.01), improve T-SOD activity and decrease the contents of MDA and ROS (P<0.05,P<0.01).Conclusion The drug-containing serum of Ficus Hirta can improve the proliferative activity of spleen lymphocyte in aged mice and the mechanism of action may be involved in decreasing the positive rate of senescent cells and increasing antioxidant ability of lymphocyte.

A novel arctigenin-containing latex glove prevents latex allergy by inhibiting type I/IV allergic reactions / 中国天然药物

The present study aimed at developing a natural compound with anti-allergic effect and stability under latex glove manufacturing conditions and investigating whether its anti-allergic effect is maintained after its addition into the latex. The effects of nine natural compounds on growth of the RBL-2H3 cells and mouse primary spleen lymphocytes were determined using MTT assay. The compounds included glycyrrhizin, osthole, tetrandrine, tea polyphenol, catechin, arctigenin, oleanolic acid, baicalin and oxymatrine. An ELISA assay was used for the in vitro anti-type I/IV allergy screening; in this process β-hexosaminidase, histamine, and IL-4 released from RBL-2H3 cell lines and IFN-γ and IL-2 released from mouse primary spleen lymphocytes were taken as screening indices. The physical stability of eight natural compounds and the dissolubility of arctigenin, selected based on the in vitro pharnacodynamaic screening and the stability evaluation, were detected by HPLC. The in vivo pharmacodynamic confirmation of arctigenin and final latex product was evaluated with a passive cutaneous anaphylaxis (PCA) model and an allergen-specific skin response model. Nine natural compounds showed minor growth inhibition on RBL-2H3 cells and mouse primary spleen lymphocytes. Baicalin and arctigenin had the best anti-type I and IV allergic effects among the natural compounds based on the in vitro pharmacodynamic screening. Arctigenin and catechin had the best physical stability under different manufacturing conditions. Arctigenin was the selected for further evaluation and proven to have anti-type I and IV allergic effects in vivo in a dose-dependent manner. The final product of the arctigenin-containing latex glove had anti-type I and IV allergic effects in vivo which were mainly attributed to arctigenin as proved from the dissolubility results. Arctigenin showed anti-type I and IV allergic effects in vitro and in vivo, with a good stability under latex glove manufacturing conditions, and a persistent anti-allergic effect after being added into the latex to prevent latex allergy.

Effect of Chuanmingshen violaceum polysaccharides and its sulfated derivatives on immunosuppression induced by cyclophosphamide in mice / 中国免疫学杂志

Objective:In oder to investigate the effect of Chuanmingshen violaceum polysaccharides ( CVP) and Solfated Chua-nmingshen violaceum polysaccharides ( SCVP) on immunosuppression induced by cyclophosphamide ( CY) in mice.Methods: CY were used to induce immunosuppression in mice;Spleen and thymus indexes were used to evaluate the immune organs indexes;the [3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltet-razolium bromide,MTT] method was used to detect the proliferation of spleen lymphocytes of each group;the concentrations of IFN-γand IL-2 were assayed by ELISA kit.Results: SCVP and CVP could resist immunosuppression by promoting lymphocyte proliferation, increasing the contents of IFN-γ and IL-2, promoting immune organs development in immunosuppressive mice induced by CY.Conclusion:SCVP and CVP exhibited the potential to used as immunopotentiator.

Antioxidant and immunoregulatory effects of Centella asiatica extracts / 中国病理生理杂志

AIM: To determine the antioxidant and the immunoregulatory effects of Centella asiatica extracts. METHODS: Centella asiatica was extracted with alcohol and different organic solvent. The content of polyphenol was determined by Folin-ciocalteau method. The efficacy of the extracts to scavenge the hydroxy radical (OH·), 1,1-diphenyl-2-picryl-hydrazyl radical (DPPH·) and intra-cellular reactive oxygen species (ROS) was measured. Lymphocyte proliferation was assessed to observe the influence of Centella asiatica extracts. RESULTS: (1) Centella asiatica was found to have abundant polyphenol extrated from different phases and in a descending order as follows: ethyl acetate extracts>n-butanol extracts>water extracts>ligarine extracts. (2) Extracts of Centella asiatica exhibited the scavenging efficacy of OH· and DPPH· free radicals, in which the acetic ether extracts showed the significant effect. (3) The acetic ether extracts had significant ability to inhibit the generation of ROS in stimulated lymphocytes. (4) The acetic ether extracts suppressed the lymphocyte proliferation. (5) The active ingredient was identified as flavone. CONCLUSION: Flavones in the Centella asiatica posseses antioxidant activity and effectively inhibits lymphocyte proliferation, showing ability of immunosuppre-ssion.

Gene expression of spleen lymphocytes induced by gp96-peptide complexes and its anti-tumor effect in vitro / 中国病理生理杂志

AIM:To study the expressing variation of TNF-? and IFN-? mRNA in mouse splenocytes induced by H22 tumor cells derived heat shock protein gp96-peptide complexes in vitro,and to observe the morphologic change of H22 tumor cells which treated with the culture supernatant.METHODS:H22 tumor cells derived HSP gp96 was obtained by the techniques for protein extraction and purification and was identified by Western blotting method.The expression values of TNF-? and IFN-? mRNA in spleen lymphocytes were detected by semi-quantitative RT-PCR.The morphologic changes of H22 tumor cells induced by the culture supernatant were observed by laser scanning confocal microscopy(LSCM)and transmission electron microscope(TEM).RESULTS:Purified heat shock protein gp96 was identified by Western blotting.The expression value of TNF-? and IFN-? mRNA in activated spleen lymphocytes induced by gp96-peptide complexes was higher than that in control groups(P