AIM: To investigate the effect of various concentrations of tetrandrine on activation of quiescent rat hepatic stellate cells (HSCs) and transforming growth factor-β (TGF-β) signaling in vitro.METHODS: HSCs were iso...AIM: To investigate the effect of various concentrations of tetrandrine on activation of quiescent rat hepatic stellate cells (HSCs) and transforming growth factor-β (TGF-β) signaling in vitro.METHODS: HSCs were isolated from rats by in situperfusion of liver and 18% Nycodenz gradient centrifugation, and primarily cultured on uncoated plastic plates for 24 hwith DMEM containing 20% fetal bovine serum (FBS/DMEM) before the culture medium was substituted with 2% FBS/DMEM for another 24 h. Then, the HSCs were cultured in 2% FBS/DMEM with tetrandrine (0.25, 0.5, 1,2 mg/L, respectively). Cell morphological features were observed under an inverted microscope, smooth muscleα-actin (α-SMA) was detected by immunocytochemistry and image analysis system, laminin (LN) and type Ⅲprocollagen (PCⅢ) in supernatants were determined byradioimmunoassay. TGF-β1 mRNA, Smad 7 mRNA and Smad 7 protein were analyzed with RT-PCR and Western blotting, respectively.RESULTS: Tetrandrine at the concentrations of 0.25-2 mg/L prevented morphological transformation of HSC from the quiescent state to the activated one, while α-SMA, LN and PCⅢ expressions were inhibited. As estimated by gray values, the expression of α-SMA in tetrandrine groups (0.25, 0.5, 1, 2 mg/L) was reduced from 21.3% to 42.2%(control: 0.67, tetrandrine groups: 0.82, 0.85, 0.96, or 0.96, respectively, which were statistically different from the control, P＜0.01), and the difference was more significant in tetrandrine at 1 and 2 mg/L. The content of LN in supernatants was significantly decreased in tetrandrine groups to 58.5%, 69.1%, 65.8% or 60.0% that of the control respectively, and that of PCⅢ to 84.6%, 81.5%,75.7% or 80.7% respectively (P＜0.05 vs control), with no significant difference among tetrandrine groups. RTPCR showed that TGF-β1 mRNA expression was reduced by tetrandrine treatments from 56.56% to 87.90% in comparison with the control, while Smad 7 mRNA was increased 1.4-4.8 times. The TGF-β1 mRNA and Smad 7 mRNA expression was in a significant negative correlation (r= -0.755, P＜0.01), and both were significantly correlated with α-SMA protein expression (r = -0.938, P＜0.01;r = 0.938, P＜0.01, respectively). The up-regulation of Smad 7 protein by tetrandrine (1 mg/L)was confirmed by Western blotting as well.CONCLUSION: Tetrandrine has a direct inhibiting effect on the activation of rat HSCs in culture. It up-regulates the expression of Smad 7 which in turn blocks TGF-β1 expression and signaling.展开更多
Bacterial quorum sensing （QS） has attracted much interests and it is an important process of cell communication. Recently, Bassler et al. studied the phenomena of QS regulated by small RNAs and the experimental data...Bacterial quorum sensing （QS） has attracted much interests and it is an important process of cell communication. Recently, Bassler et al. studied the phenomena of QS regulated by small RNAs and the experimental data showed that smafl RNAs played important role in the QS of Vibrio harveyi and it can permit the fine-tuning of gene regulation and mmntenance of homeostasis. According to Michaelis-Menten kinetics and mass action law in this paper, we construct a mathematical model to investigate the mechanism induced QS by coexist of small RNA and signal molecular （AI） and show that there are periodic oscillation when the time delay and Hill coefficient exceed a critical value and the periodic oscillation produces the change of concentration and induces QS. These results are fit to the experimental results. In the meanwhile, we also get some theoretical value of Hopf Bifurcation on time deday. In addition, we also find this network is robust against noise.展开更多
Objective ：To study curcumin＇s growth inhibitory effects and morphology changes on sarcoma graft＇s of S180 mouse, with further inquiry into the possible mechanisms, Methods： Thirty S180 mouse were randomly assigne...Objective ：To study curcumin＇s growth inhibitory effects and morphology changes on sarcoma graft＇s of S180 mouse, with further inquiry into the possible mechanisms, Methods： Thirty S180 mouse were randomly assigned into 3 groups ： saline group （blank control group）, Cytoxan group （positive control group） and curcumin group. The tumor inhibitory rates, the index of thymus and spleen,the growth of tumor and the change of pathology-morph, the index of apoptosis cells and morphology changes of apoptosis cells in the different groups were observed. Results： （1） Tumor＇s inhibitory rate in curcumin group and cytoxan group was 68, 32% and 70. 43%, respectively. Compared to blank control group, the 2 groups had significant elevated tumor inhibitory rate （P〈0.01）. （2） Thymus index of curcumin group did not have significant decrease compared to blank control group （P〉0.05）, （3） Under electroscope,curcumin group and positive control group had significant decrease in terms of growth of tumor, degree of infiltration of tumor, the number of nucleus fission, and blood vessels number compared to saline group （P〈0.05）, However, the degree of cell necrosis, the number of splenic segments and macrophage are increased significantly compared to negative group, （4） Accumulative score of apoptosis cell in curcumin group was significantly higher than other two groups（P〈0.05）. Conclusion： Internal organ study and ceil morphology observation show curcumin can effectively inhibit the growth and cause the death of sarcoma graft of S180 mouse without interference with thymus.展开更多
基金Supported by the College Science and Technology Developing Foundation of Shanghai, No. 02BK14
文摘AIM: To investigate the effect of various concentrations of tetrandrine on activation of quiescent rat hepatic stellate cells (HSCs) and transforming growth factor-β (TGF-β) signaling in vitro.METHODS: HSCs were isolated from rats by in situperfusion of liver and 18% Nycodenz gradient centrifugation, and primarily cultured on uncoated plastic plates for 24 hwith DMEM containing 20% fetal bovine serum (FBS/DMEM) before the culture medium was substituted with 2% FBS/DMEM for another 24 h. Then, the HSCs were cultured in 2% FBS/DMEM with tetrandrine (0.25, 0.5, 1,2 mg/L, respectively). Cell morphological features were observed under an inverted microscope, smooth muscleα-actin (α-SMA) was detected by immunocytochemistry and image analysis system, laminin (LN) and type Ⅲprocollagen (PCⅢ) in supernatants were determined byradioimmunoassay. TGF-β1 mRNA, Smad 7 mRNA and Smad 7 protein were analyzed with RT-PCR and Western blotting, respectively.RESULTS: Tetrandrine at the concentrations of 0.25-2 mg/L prevented morphological transformation of HSC from the quiescent state to the activated one, while α-SMA, LN and PCⅢ expressions were inhibited. As estimated by gray values, the expression of α-SMA in tetrandrine groups (0.25, 0.5, 1, 2 mg/L) was reduced from 21.3% to 42.2%(control: 0.67, tetrandrine groups: 0.82, 0.85, 0.96, or 0.96, respectively, which were statistically different from the control, P＜0.01), and the difference was more significant in tetrandrine at 1 and 2 mg/L. The content of LN in supernatants was significantly decreased in tetrandrine groups to 58.5%, 69.1%, 65.8% or 60.0% that of the control respectively, and that of PCⅢ to 84.6%, 81.5%,75.7% or 80.7% respectively (P＜0.05 vs control), with no significant difference among tetrandrine groups. RTPCR showed that TGF-β1 mRNA expression was reduced by tetrandrine treatments from 56.56% to 87.90% in comparison with the control, while Smad 7 mRNA was increased 1.4-4.8 times. The TGF-β1 mRNA and Smad 7 mRNA expression was in a significant negative correlation (r= -0.755, P＜0.01), and both were significantly correlated with α-SMA protein expression (r = -0.938, P＜0.01;r = 0.938, P＜0.01, respectively). The up-regulation of Smad 7 protein by tetrandrine (1 mg/L)was confirmed by Western blotting as well.CONCLUSION: Tetrandrine has a direct inhibiting effect on the activation of rat HSCs in culture. It up-regulates the expression of Smad 7 which in turn blocks TGF-β1 expression and signaling.
基金Supported by National Natural Science Foundation of China under Grant Nos. 10802043 and 10832006, Program for Science ＆ Technology Innovation Talents in Universities of Henan under Grant No. 2009HASTIT033 and Key Disciplines of Shanghai municipality （$30104）
文摘Bacterial quorum sensing （QS） has attracted much interests and it is an important process of cell communication. Recently, Bassler et al. studied the phenomena of QS regulated by small RNAs and the experimental data showed that smafl RNAs played important role in the QS of Vibrio harveyi and it can permit the fine-tuning of gene regulation and mmntenance of homeostasis. According to Michaelis-Menten kinetics and mass action law in this paper, we construct a mathematical model to investigate the mechanism induced QS by coexist of small RNA and signal molecular （AI） and show that there are periodic oscillation when the time delay and Hill coefficient exceed a critical value and the periodic oscillation produces the change of concentration and induces QS. These results are fit to the experimental results. In the meanwhile, we also get some theoretical value of Hopf Bifurcation on time deday. In addition, we also find this network is robust against noise.
文摘Objective ：To study curcumin＇s growth inhibitory effects and morphology changes on sarcoma graft＇s of S180 mouse, with further inquiry into the possible mechanisms, Methods： Thirty S180 mouse were randomly assigned into 3 groups ： saline group （blank control group）, Cytoxan group （positive control group） and curcumin group. The tumor inhibitory rates, the index of thymus and spleen,the growth of tumor and the change of pathology-morph, the index of apoptosis cells and morphology changes of apoptosis cells in the different groups were observed. Results： （1） Tumor＇s inhibitory rate in curcumin group and cytoxan group was 68, 32% and 70. 43%, respectively. Compared to blank control group, the 2 groups had significant elevated tumor inhibitory rate （P〈0.01）. （2） Thymus index of curcumin group did not have significant decrease compared to blank control group （P〉0.05）, （3） Under electroscope,curcumin group and positive control group had significant decrease in terms of growth of tumor, degree of infiltration of tumor, the number of nucleus fission, and blood vessels number compared to saline group （P〈0.05）, However, the degree of cell necrosis, the number of splenic segments and macrophage are increased significantly compared to negative group, （4） Accumulative score of apoptosis cell in curcumin group was significantly higher than other two groups（P〈0.05）. Conclusion： Internal organ study and ceil morphology observation show curcumin can effectively inhibit the growth and cause the death of sarcoma graft of S180 mouse without interference with thymus.
