Effects of proglumide, a gastrin receptor antagonist, on human large intestine carcinoma SW480 cell line

OBJECTIVE To explore the effects of proglumide, a gastrin receptor antagonist, on the amount of viable cells, synthesis of DNA and protein, and cell proliferation cycle in human large intestine carcinoma SW480 cell line in order to provide experimental basis for treatment of large intestine carcinoma using proglumide.

METHODS Large intestine carcinoma SW480 cells at logarithmic growth stage were cultivated with different concentrations of proglumide for different periods of time, then the amount of viable cells was determined by MTT colorimetric analysis. The SW480 cells were cultivated with proglumide, pentagastrin, proglumide + pentagastrin for the same period of time, then the contents of DNA and protein and the cell proliferation cycle were determined by flow-cytometry.

RESULTS The amount of viable cells, synthesis of DNA and protein, distribution of cell cycle, and proliferation index (PI) in the proglumide group did not differ significantly from those in the pentagastrin group (P > 0.05). The amount of viable cells in the pentagastrin group was significantly higher than that in the pentagastrin group (P < 0.01). In the proglumide + pentagastrin group the amount of viable cells, synthesis of DNA and protein, amount of S and G2M phase cells, and PI were all significantly lower than those in the pentagastrin group (all P < 0.01), and the amount of G0/G1 phase cells was significantly higher than that in the pentagastrin group (P < 0.01), but none of the above differed from those in the control group (all P > 0.05).

CONCLUSIONS Proglumide has no obvious effect on the growth of human large intestine carcinoma SW480 cell line, but can inhibit the growth-promoting effect of pentagastrin on large intestine carcinoma cells. The mechanism may be that proglumide inhibits the promoting effect of pentagastrin on the synthesis of DNA and protein of carcinoma cells, and then inhibits carcinoma cell growth from G0/G1 phase to S and G2M phase.

     

Time-effect Relationship of Extracts from Ginseng, Notoginseng and Chuanxiong on Vascular Endothelial Cells Senescence

Objective:To observe the time-effect relation of extracts from ginseng,notoginseng and chuanxiong on angiotensin II(AngⅡ)-induced senescence of vascular endothelial cells and explore the feature of Chinese medicine against vascular diseases.Methods:Human umbilical vein endothelial cells(HUVECs)cultured in vitro were stimulated with 10~(-6) mol/L Ang Ⅱ to induce cell senescence,which were divided into 4groups,the blank control group,the Ang Ⅱ model group,the extracts group and the telmisartan group.The(J-gal was used to identify senescence of cells,the cell counting kit-8 method was applied to assess the cell viability,the cell function was examined with the level of endothelial nitric oxide synthase(eNOS) and the flow cytometry was used for analyzing the cell cycle changes.Results:Compared with the control cells,the cells positive for β-gal staining was significantly increased in the Ang Ⅱ model group,and showed cell cycle arrest at G_0/G_1 phase with decreased S and G_2/M phase ceil percentage,eNOS expression and cell viability(P0.05).The extracts and telmisartan treatment of Ang II-induced cells resulted in decreased β-gal positive cells with a reduction in G_0/G_1phase cells and an increasing in S,G_2/M phase ceils and eNOS expression(P0.05).At 24 h,the extracts were more effective than telmisartan(P0.05);while telmisartan was more effective at 48 h(P0.05).Conclusion:Extracts from ginseng,notoginseng and chuanxiong can delay Ang II-induced aging of HUVECs and may play an important role in early senescence.     

Inhibitive effects of glucose and free fatty acids on proliferation of human vascular endothelial cells in vitro

Objectives To investigate the effects of glucose and free fatty acids (FFAs) on the proliferation and cell cycle of human vascular endothelial cells in vitro , and to examine whether the combined presence of elevated FFAs and glucose may cross-amplify their individual injurious effects.Methods Cultured human vascular endothelial cells (ECV304) were incubated with various concentrations of glucose and/or FFAs (palmitate and/or oleate) for 24-96 h. Morphologic alterations were observed using a phase contrast microscope and an electron microscope. Inhibition of proliferation was measured by a colorimetric 3-［4, 5-dimethyl thiazol-2-yl］-2, 5-diphenyltetrazolium bromide (MTT) assay. Cell viability was determined using trypan blue exclusion. Distribution of cells along phases of the cell cycle was analyzed by flow cytometry. Results Glucose 15 or 30 mmol/L, palmitate (PA) 0.25 or 0.5 mmol/L, and oleate (OA) 0.5 mmol/L inhibited proliferation and accelerated death of endothelial cells in a dose-and-time-dependent manner. After treatment with elevated glucose and/or FFAs, the G(0)/G(1) phase cells increased, whereas S phase cells decreased, suggesting that high glucose and/or FFAs mainly arrested endothelial cells at G(0)/G(1) phase. The inhibitive rates of proliferation and population of dead cells in endothelial cells incubated with glucose plus FFAs (glucose 30 mmol/L+PA 0.25 mmol/L, glucose 30 mmol/L+OA 0.5 mmol/L, glucose 30 mmol/L+PA 0.25 mmol/L+OA 0.5 mmol/L) increased more markedly than those treated with high glucose or FFAs (PA and/or OA) alone. Conclusion Both high ambient glucose and FFAs can inhibit proliferation and accelerate death of endothelial cells in vitro. These changes were cross-amplified in the combined presence of high levels of glucose and FFAs.     

Effects of Trichostatin A on HDAC8 Expression, Proliferation and Cell Cycle of Molt-4 Cells

The effects of Trichostatin A (TSA) on histone deacetylase 8 (HDAC8) expression, proliferation and cell cycle arrest in T-lymphoblastic leukemia cell line Molt-4 cells in vitro were investigated. The effect of TSA on the growth of Molt-4 cells was studied by MTT assay. Flow cytometry was used to examine the cell cycle. The expression of HDAC8 was detected by using immunocyto-chemistry and Western blot. The results showed that proliferation of Molt-4 cells was inhibited in TSA-treated group in a time- and dose-dependent manner. The IC50 of TSA exposures for 24 h and 36 h were 254.3236 and 199.257μg/L respectively. The cell cycle analysis revealed that Molt-4 was mostly in G0/G1 phase, and after treatment with TSA from 50 to 400μg/L for 24 h, the percents of G0/G1 cells were decreased and cells were arrested in G2/M phase. Treatment of TSA for 24 h could significantly inhibit the expression of HDAC8 protein in Molt-4 cells (P<0.01). It was concluded that TSA could decrease the expression of HD AC8 in Molt-4 cells, which contributed to the inhibition of proliferation and induction of cell cycle arrest in Molt-4 cells.     

Up-regulation of HIF-1α and VEGF Expression by Elevated Glucose Concentration and Hypoxia in Cultured Human Retinal Pigment Epithelial Cells

Summary： In order to explore the effect of high glucose concentration and high glucose concentration with hypoxia on the production of hypoxia-inducible factor-1α （HIF-1α） and vascular endothelial growth factor （VEGF）, human RPE cells were cultured in 5,56 mmol/L glucose （control group）, 5.56 mmol/L glucose with 150 !a mol/L COCl2 （hypoxic group）, 25 mmol/L glucose （high glucose group） and 25 mmol/L glucose with 150 μmol/L COCl2 （combination group）. RT-PCR was used to detect the expression of HIF-1α and VEGF mRNAs. Western blot analysis was used to measure the levels of HIF-1α and VEGF proteins. Although the small amount of HIF-1α protein was able to be detected in high glucose group but not in control group, there was no significant difference between the expression of HIF-1α mRNA of RPE cells in high glucose group and that of RPE cells in control group. As compared with RPE cells in control group, the mRNA expression and the protein synthesis of VEGF in high glucose group were up-regulated. As compared with RPE cells in hypoxic group, the expression of HIF-1α mRNA of RPE cells in combination group was not different, but the protein synthesis of HIF-1α, the mRNA expression and the protein synthesis of VEGF were more obviously up-regulated. In conclusion, high concentration glucose mainly influence the protein synthesis of HIF-1α of RPE cell, and HIF-1α protein is able to be accumulated in high concentration glucose. Under hypoxia, the HIF-1α protein induced by high concentration glucose is more stable, and the expression of VEGF is obviously increased. It is suggested that high concentration glucose may play a role in retinal neovascularization, especially at ischemia stage of diabetic retinopathy.     

筋脉通胶囊血清有效成分可预防大鼠高血糖浓度引起的施万细胞凋亡

Objective: To evaluate the effect of serum containing Jinmaitong Capsule (筋脉通胶囊, JMT) on apoptosis of Schwann cells (SCs) that are cultured in high glucose at the cellular and molecular levels. Methods: SCs were cultured in Dulbecco''s modi?ed Eagle''s medium (control group), high glucose (50 mmol/L) medium supplemented with 20% rat serum (HG group), and 50 mmol/L glucose medium supplemented with serum containing JMT (JMT group). SC apoptosis was detected using a terminal deoxynucleotidyl transferase dUTP nick end labeling kit. The expression of Bcl-2 and the caspase-3 p20 subunit in SCs were detected by real-time fluorogenic quantitative polymerase chain reaction and confocal laser scanning microscopy, respectively. Results: No apoptosis was detected in SCs that were cultured in the control group. The percentage of apoptosis of SCs cultured in the HG group was much higher than that in the control group. The apoptosis of SCs in the JMT group was lower than that in the HG group. Fluorescence intensity of Bcl-2 and the expression of Bcl-2 mRNA in SCs that were cultured in the HG group were much lower than those in the control group and much higher than those in the JMT group (P<0.01). The fluorescence intensity of caspase-3 p20 and the expression of caspase-3 p20 mRNA in SCs that were cultured in the HG group were much higher than those in the control group (P<0.01), and they were remarkably lower in the JMT group (P<0.01). Conclusions: JMT effectively prevents SC apoptosis that is induced by high glucose. This effect may be because of increased expression of Bcl-2 mRNA and protein and decreased expression of caspase-3 p20 mRNA and protein.     

Role of transforming growth factor-beta in the growth inhibition of cultured rat mesangial cells in high glucose media.

The present study was designed to test the hypothesis that the inhibitory effect of elevated glucose levels on mesangial cell growth might be mediated by transforming growth factor beta (TGF beta). Increased glucose levels inhibited mesangial cell proliferation in a concentration dependent manner. The addition of a rabbit antipocine TGF beta 1 neutralizing antibody significantly enhanced the cell proliferation in both normal (5.5 mM) and high (50 mM) glucose media (12% and 30% respectively), and almost completely blocked the growth inhibition induced by high glucose media. TGF beta assay demonstrated mesangial cells produced more active than latent TGF beta after growth in high glucose media. We conclude that TGF beta functions as an autocrine cytokine in mesangial cells growth and the production of TGF beta is modulated by high glucose concentration. The growth inhibition induced by high glucose levels may be largely mediated by endogenous TGF beta.

     

Trichostatin A Regulates hGCN5 Expression and Cell Cycle on Daudi Cells in vitro

The expression of human general control of amino acid synthesis protein 5 (hGCN5) in human Burkitt's lymphoma Daudi cells in vitro, effects of Trichostatin A (TSA) on cell proliferation and apoptosis and the molecular mechanism of TSA inhibiting proliferation of Daudi cells were investigated. The effects of TSA on the growth of Daudi cells were studied by 3-(4, 5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium (MTT) assay. The effect of TSA on the cell cycle of Daudi cells was assayed by a propidium iodide method. Immunochemistry and Western blot were used to detect the expression of hGCN5. The proliferation of Daudi cells was decreased in TSA-treated group with a 24 h IC50 value of 415.3979μg/L. TSA induced apoptosis of Daudi cells in a time- and dose-dependent manner. Treatment with TSA (200 and 400μg/L) for 24 h, the apoptosis rates of Daudi cells were (14.74±2.04) % and (17.63±1.25) %, respectively. The cell cycle was arrested in G0/G1 phase (50, 100μg/L) and in G2/M phase (200μg/L) by treatment with TSA for 24 h. The expression of hGCN5 protein in Daudi cells was increased in 24 h TSA-treated group by immu-nochemistry and Western blot (P<0.05). It was suggested that TSA as HDACIs could increase the expression of hGCN5 in Daudi cells, and might play an important role in regulating the proliferation and apoptosis of B-NHL cell line Daudi cells.     

Effect and Mechanism of Epidermal Growth Factor on Proliferation of GL15 Gliomas Cell Line

The effects of epidermal growth factor (EGF) on proliferation of G15 glioma cells and the possible mechanisms were investigated. GFAP and EGFR expression was detected by immunohisto-chemical method. After the cells were treated with EGF at different concentrations, cell count method was used to determine the proliferation of glioma cells, cell cycle and apoptosis were analyzed by flow cytometry (FCM), and laser scan confocal microscope (LSCM) was used to measure the cyto-plasmic free calcium. The results showed that GFAP was diffusedly expressed in GL15 cells and EGFR was over-expressed. EGF at doses of≤1 ng/mL could significantly stimulate cell proliferation, cells in phase G0/G1 decreased, and those in phase S increased. EGF at doses of 10 and 100ng/ml could inhibit the cell proliferation significantly, and the apoptosis ratio in high dose of EGF group was higher than in control group. EGF could significantly induce a quick rise of intracellular free calcium, but the peak value of intracellular free calcium activated by high dose of EGF was higher than by low dose of EGF. It was suggested that EGF had a dual effect on gliomas: low dose of EGF could stimulate the cell proliferation of gliomas, but high dose of EGF could induce the cell apoptosis and inhibit the proliferation of gliomas, which might be contributed to the difference of intracellular free calcium.     

叶绿酸铜钠对再生障碍性贫血小鼠间充质干细胞功能的影响

Objective: To investigate the effects of sodium copper chlorophyllin (SCC) on the proliferation, differentiation and immunomodulatory function of mesenchymal stem cells (MSCs) from mice with aplastic anemia. Methods: A mouse model of aplastic anemia was established by exposure of BALB/c mice to sublethal doses of 5.0 Gy Co60 γ radiation, followed by transplantation of 2×1000000 lymph node cells from DBA/2 donor mice within 4 h after radiation. Aplastic anemic BALB/c mice were randomly divided into six groups: the treated groups, which received 25, 50, or 100 mg/kg/day SCC, respectively; a positive control group treated with cyclosporine A (CsA); and an untreated model control group (model group); while, the non-irradiated mice as the normal control group. SCC or CsA were administered by gastrogavage for 20 days, starting on day 4 after irradiation. Peripheral blood cells were counted and colony-forming fibroblasts (CFU-F) in the bone marrow were assayed. The ability of MSCs to form calcium nodes after culture in osteoinductive medium was also observed. The immunosuppressive effect of MSCs on T lymphocytes was analyzed by enzyme-linked immunosorbent assay and flow cytometry, to evaluate the efficacy of SCC in mice with aplastic anemia. Results: Peripheral blood white cell and platelet counts were increased by medium and high SCC doses, compared with the untreated control. CFU-Fs were also increased compared with the untreated control, and the numbers of calcium nodes in MSCs in osteoinductive medium were elevated in response to SCC treatment. The percentage of Forkhead box protein 3 (FOXP3+) T cells was increased in T cell-MSC cocultures, and the cytokine transforming growth factor β1 was up-regulated in SCC-treated groups. Conclusion : The results of this study suggest that SCC not only promotes the proliferation and differentiation of MSCs, but also improves their immunoregulatory capacity in mice with aplastic anemia.     

Effects of suppressed autophagy on mitochondrial dynamics and cell cycle of N2a cells

Autophagy dysregulation, mitochondrial dynamic abnormality and cell cycle re-entry are implicated in the vulnerable neurons of patients with Alzheimer＇s disease. This study was designed to testify the association among autophagy, mitochondrial dynamics and cell cycle in dividing neuroblas- toma （N2a） cells. The N2a cells were cultured in vitro and treated with different concentrations of 3-methyladenine （3-MA）. The cell viability was detected by methyl thiazolyl tetrazolium （MTT） assay. They were randomly divided into control group （cells cultured in normal culture medium） and 3-MA group （cells treated with 10 mmol/L 3-MA）. The cell cycle was analyzed in the two groups 3, 6, 12, and 24 h after treatment by flow cytometry. Western blotting was used to evaluate the expression levels of mitofission 1 （Fisl）, mitofusin 2 （Mfn2）, microtubule-associated protein 1 light chain 3 （LC3）, cell cy- cle-dependent kinase 4 （CDK4） and cdc2. The flow cytometry revealed that the proportion of cells in G2/IVI was significantly increased, and that in G0/G1 was significantly reduced in the 3-MA group as compared with the control group. Western blotting showed that the expression levels of Fisl, LC3, and CDK4 were significantly up-regulated in the 3-MA group at the four indicated time points as compared with the control group. Mfn2 was initially decreased in the 3-MA group, and then significantly in- creased at 6 h or 12 h. Cdc2 was significantly increased in the 3-MA group at 3 h and 6 h, and then dropped significantly at 12 h and 24 h. Our data indicated that 3-MA-induced suppressed autophagy may interfere with the cell cycle progression and mitochondrial dynamics, and cause cell death. There are interactions among cell cycle, mitochondrial dynamics and autophagy in neurons.     

Effect of taurine on GFAP and TauT expressions in rat retinal Müller cells in high glucose culture

Objective： To detect the expression of glial fibrillary acid protein （GFAP） and taurine transporter （TauT） in the retinal Müller cells in high glucose culture with taurine and to explore the influence of glucose on the taurine transporting, and the possible protective effects of taurine on MUller cells in early diabetic retinopathy. Methods： The Müller cells from the rat retina were cultured in high glucose, and GFAP and Taut expressions were detected in the cells treated with different doses of taurine by immuocytochemical fluorescein staining and Western blotting. Results： High glucose enhanced the expression of GFAP and decreased the expression of TauT in Müller cells. Taurine decreased the up-regulation of GFAP in the cells which was induced by high glucose; 0. 1-10 mmol/L taurine increased the expression of TauT in Müller cells. Conclusion： Taurine can inhibit the changes in Müller cell resulted from high glucose.     

Human vascular smooth muscle cells and endothelial cells cocultured on polyglycolic acid (70/30) scaffold in tissue engineered vascular graft

Background Current prosthetic, small diameter vascular grafts showing poor long term patency rates have led to the pursuit of other biological materials. Biomaterials that successfully integrate into surrounding tissue should match not only the mechanical properties of tissues, but also topography. Polyglycolic acid (70/30) has been used as synthetic grafts to determine whether human vascular smooth muscle cells and endothelial cells attach, survive and secrete endothelin and 6-keto-prostaglandin F1α (6-keto-PGF1α).Methods Endothelial cells and smooth muscle cells were isolated from adult human great saphenous vein. They were seeded on polyglycolic acid scaffold in vitro separately to grow vascular patch (Groups A and B respectively) and cocultured in vitro to grow into vascular patch (Group C). Smooth muscle cells and endothelial cells were identified by immunohistochemical analysis and growth of cells on polyglycolic acid was investigated using scanning electron microscopy. The levels of endothelin and 6-keto-PGF1α in the culturing solutions were examined by radioimmunology to measure endothelial function. Results Seed smooth muscle cells adhered to polyglycolic acid scaffold and over 28 days grew in the interstices to form a uniform cell distribution throughout the scaffold. Then seed endothelial cells formed a complete endothelial layer on the smooth muscle cells. The levels of endothelin and 6-keto-prostaglandin F1 alpha in the culturing solution were (234±29) pg/ml and (428±98) pg/ml respectively in Group C and (196±30) pg/ml and (346±120) pg/ml in Group B; both significantly higher than in Groups A and D (blank control group, all P&lt;0.05 ). Conclusions Cells could be grown successfully on polyglycolic acid and retain functions of secretion. Our next step is to use human saphenous vein smooth muscle cells and endothelial cells to grow tubular vascular grafts in vitro.     

Over-expression of LRIG3 suppresses growth and invasion of bladder cancer cells

The purpose of this study was to investigate the impact of leucine-rich repeats and immunoglobulin-like domains 3 (LRIG3) on the biological features of bladder cancer cell lines. The plasmids of over-expressed LRIG3 and the blank plasmid serving as control were transfected into the bladder cancer cell lines, T24, EJ and BIU-87, and the expression levels of LRIG3 mRNA and protein were detected by using real-time PCR and Western blotting. The changes in the cell cycle and apoptosis were examined by using flow cytometry. The invasive ability was measured by Transwell assay, and CCK-8 assays were used to measure the proliferation of cells. As compared with the control group, the LRIG3 mRNA and protein expression levels in LRIG3 cDNA-transfected group were raised significantly (P<0.05). The average number of cells with up-regulated LRIG3 passing through the inserted filter was decreased significantly as compared with the control group (P<0.05). Up-regulation of LRIG3 also could inhibit proliferation and induce apoptosis of T24, EJ and BIU-87 cells. Except BIU-87, the T24 and EJ cells transfected with LIRG3 cDNA were arrested in G 0 /G 1 phase compared to the control group (P<0.05). In conclusion, the over-expression of LRIG3 could influence the cell cycle and invasion, inhibit proliferation and induce apoptosis in the three bladder cancer cell lines.     

Effects of glucose and insulin on the H9c2 （2-1） cell proliferation may be mediated through regulating glucose transporter 4 expression

Background The change of glucose transporter 4 （GLUT4） expression could influence glucose uptake in the myocardial cells and then effect myocardial metabolism, which maybe one of the factor for the diabetes cardiovascular disease. This study aimed to explore the influence of glucose and insulin at different concentrations on H9c2 （2-1） cell proliferation and its GLUT4 expression in vitro, and evaluate the correlation between myocardial cells proliferation and GLUT4 expression. This might be helpful for understanding the relationship between glucose metabolism and cardiovascular disease. Methods According to glucose concentrations in culture medium, cultured H9c2 rat myocardial cells were divided into five groups： control group （NC, glucose concentration 5.0 mmol/L）, low glucose group （LG, glucose concentration 0.1 mmol/L）, high glucose group 1 （HG1, glucose concentration 10 mmol/L）, high glucose group 2 （HG2, glucose concentration 15 mmol/L）, high glucose group 3 （HG3, glucose concentration 20 mmol/L）. Then according to different insulin concentrations in culture medium, each group was further divided into two subgroups： normal insulin subgroup （INSc, insulin concentration 3.8 mU/L）, high insulin subgroup （INSh, insulin concentration 7.6 mU/L）. H9c2 （2-1） cells were cultured for 1, 2, 3 days, the proliferation of cells were assayed by cell counting Kit-8 assay, the expressions of GLUT4 mRNA and protein were detected with RT-PCR and Western Blotting technique, and the relation between myocardial cells proliferation and GLUT4 expression was evaluated.     

Effect of taurine on GFAP and TauT expressions in rat retinal Müller cells in high glucose culture

Objective： To detect the expression of glial fibrillary acid protein （GFAP） and taurine transporter （TauT） in the retinal Müller cells in high glucose culture with taurine and to explore the influence of glucose on the taurine transporting, and the possible protective effects of taurine on MUller cells in early diabetic retinopathy. Methods： The Müller cells from the rat retina were cultured in high glucose, and GFAP and Taut expressions were detected in the cells treated with different doses of taurine by immuocytochemical fluorescein staining and Western blotting. Results： High glucose enhanced the expression of GFAP and decreased the expression of TauT in Müller cells. Taurine decreased the up-regulation of GFAP in the cells which was induced by high glucose; 0. 1-10 mmol/L taurine increased the expression of TauT in Müller cells. Conclusion： Taurine can inhibit the changes in Müller cell resulted from high glucose.     

Resveratrol Inhibits the Secretion of Vascular Endothelial Growth Factor and Subsequent Proliferation in Human Leukemia U937 Cells

This study examined the effect of resveratrol on the secretion of vascular endothelial growth factor(VEGF) and subsequent proliferation of human leukemia U937 cells,and explored the mechanisms involved.Human leukemia U937 cells were treated with resveratrol of different concentrations(12.5-200 μmol/L) for different time lengths(12-48 h).The proliferation of the U937 leukemic cells was determined by MTT assay.Apoptosis was observed by Annexin-Ⅴ-FIFC/PI double staining and flow cytometry(FCM).Cells cycle was analyzed by PI staining and FCM.The content of VEGF was determined by ELISA.Human umbibical vein endothelial cells were examined for vasoformation in vitro after exposures to resveratrol of various concetrations.The results showed that resveratrol inhibited the proliferation of U937 leukemia cells in a dose-and time-dependent manner.Resveratrol induced apoptosis and S-phase cell cycle arrest in human leukemic U937 cells.Resveratrol inhibited the secretion of VEGF in U937 cells.Resveratrol inhibited the vasoformation of human vein endothelial cells in a dose-dependent manner.It was concluded that resveratrol could down-regulate the secretion of VEGF,induce apoptosis and suppress the proliferation of U937 cells.     

温化蠲痹方对于胶原蛋白性关节炎大鼠滑膜细胞增殖和凋亡的作用

Objective: To study the effect of Wenhua Juanbi Recipe (温化蠲痹方, WJR) on proliferation and apoptosis of synoviocytes in rats with collagen-inducing arthritis (CIA). Methods: A CIA model was induced by intradermal injection of bovine collagen type Ⅱ emulsion at the base of rat tails. Thirty modeled healthy Wistar rats were randomly assigned to one of three groups (10 per group): the model group, the methotrexate (MTX)-treated group (0.78 mg/kg) and the WJR-treated group (22.9 g/kg). A group of 10 healthy rats was used as normal control. Treatments or normal saline for the control group were administered by oral gavage once daily. Rats were sacrificed after 30-day treatment and subjected to the following examinations: arthritis index (AI) was estimated, inflammatory cell infiltration and proliferation in synovial membrane were evaluated by microscopy, the synoviocyte apoptosis was determined by TUNEL assay, and the cell apoptosis index was calculated. Results: AI was lowered significantly in the WJR group compared to the model group (P<0.01). The pathological findings observed in the model group were reversed in the WJR group, including increase in inflammatory cell infiltration and synoviocyte proliferation in synovial membrane and reduction in cell apoptosis index (all P<0.01). Conclusions: Synoviocyte proliferation and apoptosis reduction were present in CIA rats. WJR was effective in treating the rat model of CIA. The therapeutic effect might be exerted through inducing apoptosis and suppressing proliferation of synoviocytes.