Induction of tumor-specific cytotoxic T lymphocytes in vitro by dendritic cells pulsed with different glioma antigens

AIM: The goal of this study was to compare different methods for tumor antigen preparation, to observe the induction of tumor-specific cytotoxic T lymphocytes in rats by dendritic cells (DCs) pulsed with different tumor antigens. METHODS: The precursors of dendritic cells were isolated from bone marrow of rats, stimulated in vitro with recombinent rat granulocyte-macrophage colony-stimulating factor (rrGM-CSF) and interleukin-4 (rrIL-4). Then rat DCs were pulsed with C6 tumor cell antigens prepared with different methods: freeze-thaw, boiling or total protein extracted from ultrasonic crushed tumor cell. Subsequently primed DCs were cocultured with T lymphocytes isolated from spleen to induce CTL. Lymphocyte chemoattractant factor from DCs and cytokine IFN-γ release were determined by ELISA, the cytotoxicity of CTL was assayed by JAM test. RESULTS: DCs pulsed with boiled tumor cell in vitro induced an enhanced ability of T-cell proliferation and cytotoxic T lymphocyte activity.CONCLUSION: Our results demonstrated that DCs primed with boiled tumor cell may represent a method for inducing immune responses against the entire repertoire of tumor antigens of malignancies.     

Effects of changing PKC activity on proliferation and telomerase expression in human hepatocellular carcinoma cells

AIM: To investigate whether protein kinase C (PKC) is involved in the proliferation and the telomerase expression in human hepatocellular carcinoma cells. METHODS: Human hepatocellular carcinoma cells (BEL-7402) were treated with exogenous phorbol-12-myristate-13-acetate (PMA, PKC activator) and staurosporine (SP, PKC inhibitor) for 48 hours. The techniques of cell culture and the telomeric repeat amplification protocol silver staining in combination with computer image scanning system in vitro were used to observe the variations of the growth and the telomerase expression. RESULTS: The proliferative potential of BEL-7402 cells was decreased by the action of PMA as well as SP, and the telomerase expression was also inhibited by PMA and SP. CONCLUSION: Our findings suggest that the proliferation of human hepatocellular carcinoma cells and the telomerase expression may be related to PKC.     

The mechanism of apoptosis induced by homoharringtonine in HL-60 cells

AIM: To study the signal transduction pathway of apoptosis initiation induced by homoharringtonine in HL-60 cells. METHODS: After establishing the model of apoptosis initiation induced by homoharringtonine in HL-60 cells, at the point of apoptosis initiation, molecular caspase-3, Bcl-2, Bax and Fas/FasL were measured with flow cytometry and transmission electron microscope. ERK2 and P38 expression in HL-60 cells were detected by using immunohistochemistry. RESULTS: The model of apoptosis initiation induced by homoharringtonine was established in HL-60 cells. At the point of apoptosis initiation, upregulation of caspase-3 and decrease in Bcl-2/Bax were observed. However, the expression of Fas/FasL did not significantly change. ERK2 expression decreased and P38 expression increased. CONCLUSIONS: Caspase-3, Bcl-2, Bax and mitogen activated protein kinase pathways were involved in signal transduction of apoptosis initiation induced by homoharringtonine in HL-60 cells.     

Effects of component of some Chinese herbs on proliferation of human umbilical vein endothelial cells in vitro

AIM: To observe the effects of some component of Chinese herbs for external use on proliferation of human umbilical vein endothelial cells (HUVEC) and investigate the mechanism of promoting tissue repair. METHODS: The method of MTT was used to examine the effects of Rg1, Rh1, perlolyrine, cinnamyl aldehyde, muscone, astragaluspolysaccharin (APS), velver antler polypeptide (VAP) and soluble extract of boswellia carterii birdw (BCB) on proliferation of HUVEC. RESULTS: APS did not promote proliferation of HUVEC at 9.75 mg/L-2.5 g/L; Rh1 promoted proliferation of HUVEC at 1.94 mg/L-0.5 g/L (P<0.05 or P＜0.01), and Rg1 inhibited proliferation of HUVEC at 31 mg/L (P<0.05); VAP promoted proliferation of HUVEC at 1 mg/L-0.5 g/L with optimal dose of 10 mg/L (P<0.01), Cinnamyl aldehyde promoted proliferation of HUVEC at 2 g/L(P＜0. 05); Muscone and soluble extract of BCB inhibited proliferation of HUVEC at 1 g/L, 0.5-2.5 kg/L(P＜0. 01), respectively; Perlolyrine inhibited proliferation of HUVEC at 0.125 g/L-0.5 g/L(P＜0. 01). CONCLUSION: The external herbs for supplementing Qi and warming Yang can promote HUVEC proliferation and improve angiogenesis during tissue repair. The external herbs for promoting blood circulation and accelerating capillary movement may have influence upon other stages of tissue repair.     

Gene expression of 5α-reductase (type 2) in reproductive gland of pubertal diabetic rats

AIM: To study the molecular mechanism of reproduction dysfunction in pubertal diabetic rat, the level of 5α-reductase (type 2) mRNA in testis, epididymis and prostate in diabetic rat was detected. METHODS: The gene expression of 5α-reductase (type 2) was detected by Northern blot. RESULTS: In the caput of the epididymis, the expression of 5α-reductase (type 2) mRNA of D and ID groups was less than that of C group. CONCLUSION: The decreased expression of 5α-reductase (type 2) results in the decreased production of dihydrotestosterone, which influences the development and function of reproduction system in pubertal rats.     

The molecular mechanism of inhibition of murine Lewis lung carcinoma metastasis by weimaining in vivo

AIM: To investigate the anti-metastasis effect of weimaining, extracted from fragopyrum cymosum meissn, a Chinese medicine, on murine Lewis lung carcinoma (3LL). METHODS: The anti-metastasis effect of weimaining in vivo was detected in the grafting lung metastasis model of murine Lewis lung carcinoma. The effects of the drug on the expression of CD34 and E-cadherin were investigated by immunohistochemical staining and RT-PCR. RESULTS: Weimaining effectively inhibited the lung metastasis of 3LL at a concentration of 250 mg·kg^-1·d^-1, significantly suppressed the expression of CD34 and increased the expression levels of E-cadherin protein and mRNA in 3LL cells. CONCLUSIONS: Weimaining inhibits the metastasis of murine Lewis lung carcinoma (3LL) in vivo via increasing the expression of E-cadherin and decreasing microvessel density of tumor tissue.     

Effects of Sini decoction on the ultrastructure of small intestinal epithelial cells in rats with intestinal ischemia-reperfusion

AIM: To investigate the effect of Sini decoction (SND)on the ultrastructure of small intestinal epithelial cells in rats with intestinal ischemia-reperfusion. METHODS: Thirty-two Sprague-Dawley rats of both sexes were randomly divided into 4 groups: (1) Control group in which sham operation was performed; (2) Model group in which intestinal I/R was produced by clamping super mesenteric artery(SMA) for 1 hour and declamping SMA for 3 hours; (3) SND1 group in which SD (0.6 g/200 g rat) was given via stomach tube 3 d before intestinal I/R; (4) SND2 group in which SD (1.2 g/200 g rat)was given via stomach tube 3 d before intestinal I/R. A strip of small intestine was taken from distal end of ileum for electron microscopic examination. The two-dimensional structural parameters and three-dimensional structural parameters of mitochondria were calculated. RESULTS: (1)Morphological changes of small intestine: In control group, epithelial cells were orderly arranged, with normal mitochondria and intestinal villi. In model group, the gaps between epithelial cells widened. There were a lot of apoptotic cells. Microvilli were short and swelled. Mitochondria were swelled obviously with broken ridges. Endoplasmatic reticulum was severely dilated. In SND1 and SND2 groups, microvilli and epithelial cells were orderly arranged relatively, mitochondria was slightly swelled. (2) Structural parameters of mitochondria: In model group, there were the least mitochondria and the swelling of mitochondria was severe. In SND1 and SND2 groups, the mitochondria was more than that of model group and the swelling were slight. CONCLUSION: Sini decoction can protect small intestine from ischemia-reperfusion injury without dose-dependent effect.     

Effect of YIGU capsule on IGF-I mRNA and protein expression in rat osteoblasts in vitro

AIM: The effects of YIGU capsule on proliferation and IGF-I mRNA protein expressions in osteoblasts were studied. METHODS: (1) Forty 12-month old Sprague-Dawley female rats were divided randomly into four groups (YIGU capsule high dose group, medium dose group and low dose group; saline group), the drug-containing serum and control serum were prepared. (2) The new-born Sprague-Dawley rat osteoblasts were cultured with different YIGU capsule drug-containing serum at different concentrations and different exposure time. MTT method was used to observe proliferation of osteoblasts. (3) RT-PCR method was used to measure the relative IGF-I mRNA levels and ELISA method was used to measure IGF-I secretion at different exposure time. (4) ELISA method was used to measure IGF-I secretion at different exposure time. RESULTS: (1) Proliferation of osteoblasts was more than the control groups after 48, 72 and 96 h, respectively (P<0.01); (2) The relative IGF-I mRNA levels and IGF-I protein expression were higher than those in control group after 48, 72 and 96 h, respectively (P<0.01 or P<0.05). CONCLUSIONS: It was suggested that YIGU capsule drug-containing serum promoted proliferation, IGF-I mRNA and protein expression. These results may be parts of the mechanisms of YIGU capsule to prevent and treat osteoporosis.     

Long-term effects of TCV116 on left ventricular remodeling and heart function after myocardial infarction in rats

AIM: To investigate the effects of long-term TCV116 on left ventricular remodeling and heart function after myocardial infarction. METHODS: Myocardial infarction (MI) was caused by ligation of the left anterior descending coronary artery in rats. One week after the surgical performance, the surviving rats were randomly assigned to the following treatment protocols: (1) MI rats with no therapy; (2) MI rats treated with TCV116 2 mg/kg per day; (3) Sham-operated control; (4) Sham-operated rats, treated with TCV116 2 mg/kg per day. At 22 weeks, cardiac hemodynamic parameters such as MAP, LVSP, dp/dt_max and LVEDP, and histomorphometric parameters such as LVW/BW and LVCA/BW were measured, mRNA of cardiac genes such as βMHC, BNP, TGF-β₁, collagen I and III were quantified, and survival rates were calculated. RESULTS: Compared with sham-operated rats, MI rats without therapy showed significant increases in histomorphometric parameters as well as in mRAN expressions of cardiac genes (P<0.01); While their hemodynamic parameters were significantly impaired (P<0.01), and survival duration shortened (P<0.05). Compared with MI rats without therapy, MI rats treated with TCV116 showed significant attenuation of mRAN expression of cardiac genes (P<0.01); While their hemodynamic parameters were significantly improved (P<0.05 or P＜0.01), and survival duration extended (P<0.05). CONCLUSION: Treatment with long-term angiotensin II type 1 receptor antagonist may improve left ventricular remodeling and cardiac function after MI in rats.