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Xiamen Everbeen Magnet Electron Co., Ltd.

Add: Unit H, 4F Beirongxin Mansion, No. 8 Xinfeng 2nd road, Torch Hi-Tech Zone, Xiamen, China.

Tel: 0086-592-5781916

Fax: 0086-592-5123653

E-mail:
sales@china-magnet.net; info@china-magnet.net

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Rare-earth ions on NIH3T3 and MC3T3 cell proliferation and the role of potassium channels in cell membranes

Time:06/03/2011

Rare-earth ions on NIH3T3 and MC3T3 cell proliferation and the role of potassium channels in cell membranes

Everbeen magnets supply you the rare earth magnets such as neodymium magnets,ceramic magnets,which is the strong magnets and permanent magnets.and rubber magnets,fridge magnets,Refrigerator Magnets etc.
Xiamen Everbeen Magnet Electron Co.,Ltd. http://www.chinamagnets.biz
Add:Unit H, 4F Rihua Mansion, No. 8 Xinfeng 2nd road, Torch Hi-Tech Zone, Xiamen, China.
Tel:0086-592-5781916
Fax:0086-592-5123653
E-mail:sales@china-magnet.net

With the increasingly widespread application of rare earth, especially in a large number of rare earth has been used in agriculture and husbandry, the rare earth is more and more into the environment, enter the food chain, so their physiology, toxicology, pharmacology has attracted wideattention.Meanwhile, the biological effects of research has been a hot spot.Rare earth compounds with a variety of biological activity and drug activity, but its exact mechanism has not yet fully understood, has limited the application of rare earth reasonable safety.Ion channels in non-excitable cells differentiation, growth and development plays an important role, however, can be excited on the rare earth of non-proliferation and the relationship of ion channels are few.Therefore, ion channels and explore the relationship between cell proliferation and differentiation and its mechanism is of great significance.By using whole cell patch clamp technique of rare earth lanthanum and ytterbium on cultured rat fibroblast NIH3T3 cells and rat osteoblast MC3T3 cell outward potassium current and the role of dynamic processes, using methods of cell biology of La and Ybon cultured rat fibroblast NIH3T3 cells and MC3T3 osteoblasts proliferation, apoptosis and cell cycle, to reveal the effects of potassium channel in the rare earth in the process of cell proliferation and its molecular mechanism.The main results are as follows:

MTT method and flow cytometry results showed that: (1) La3 + can significantly promote the growth of fibroblast NIH3T3 cells, and has a certain amount of time and concentration dependent manner.100 State of La3 + for 72 hours after the proliferation rate reached 46.7%.(2) At the same time, Ca2 + can also be time and concentration-dependent manner to promote the proliferation of NIH3T3 cells, and La3 + and Ca2 + in promoting proliferation of a synergistic effect.These instructions of lanthanum on the role of NIH3T3 cell proliferation associated with calcium ions and calcium ions on cell proliferation is essential.(3) La3 + also promote the proliferation of osteoblastic MC3T3 cells, a certain amount of time and concentration dependent manner.50μM of La3 + for 72 hours on the cell proliferation rate reached 38.7%.(4) In contrast with La3 +, Yb3 + cells inhibited the growth of NIH3T3 fibroblasts, and has a certain amount of time and concentration dependent manner.The role of Yb3 + 1mM NIH3T3 cell inhibition rate after 72 hours can reach 57.2%.(5) and Yb3 + on the osteoblast MC3T3 cell growth is basically not much affected.In addition, (6) in the selected dose and time, the rare earth La3 + and Yb3 + on the apoptosis of NIH3T3 cells and MC3T3 cells had no significant effect.

Cell cycle flow cytometry results showed that: (1) the role of different concentrations of La3 + NIH3T3 cells after 72h, compared with the control group, La3 + to significantly reduce the number of Gl phase cells, S phase cells increased significantly, and have some timeand concentration dependent manner.100μM of La3 + to S phase cells increased from 6.6% to 26.1%.That La3 + by promoting the G1 / S cells in the process of promoting cell proliferation.(2) different concentrations of Ca2 +; different concentrations of La3 + and 5mM of Ca2 + in the cell cycle results compared with control, Ca2 + to G1 phase cells have a certain reduction, S phase cells have a certain increase, but not very clear; La3 + and Ca2 + to the number of G1 phase cells have a more significant reduction, S phase cells have a more significant increase.The results suggest synergistic effects of lanthanum and calcium, 1000M of La3 +, Ca2 + and 5mM to S phase cells increased from 26.1% to 30.6%.(3) different concentrations of La3 + effect of MC3T3 cells 72h, rare earth La3 + on MC3T3 cell cycle had no significant effect, indicating that La3 + promotion NIH3T3 cells and MC3T3 cell proliferation mechanism is not entirely consistent, probably due to different types of cells resulting from differencesThe.(4) different concentrations of Yb3 + the role of NIH3T3 cells in 72h, on the NIH3T3 cell cycle was not significant, the G2 / M cells, the percentage slightly increased, combined with its on NIH3T3 cell proliferation that, Yb3 + may cause cell cycle arrestat the G2 / M phase, thereby inhibiting cell growth.(5) different concentrations of Yb3 + to their cell cycle compared with the control group, no difference.

Whole-cell patch-clamp study Ⅰ: (1) NIH3T3 cell outward potassium current with the calcium-dependent, and their half of the increased concentration of EC50 was 252 nmol / L.However, compared with excitable cells, the current density is much smaller.This may be a non-excitatory NIH3T3 cells or a small number of potassium channel open probability of potassium channels caused by a small reason.(2) both La and Yb rare earth ions are concentration-dependent manner inhibits fibroblast NIH3T3 cell outward potassium current, EC50 values were 1.58 and 10.63μmol / L, because the EC50 values of inhibition means that the stronger the smallerTherefore, rare earth ions La NIH3T3 rat fibroblast cells inhibition of outward potassium current is stronger than that ytterbium ions.(3) both La and Yb rare earth ions on the potassium current inhibition kinetic analysis, we found that lanthanum half of the activation of potassium currents shifted to positive potential voltage of 16mV, the potassium current to the negative half inactivation voltagepotential direction 9.48mV.On the contrary, ytterbium ions to outward potassium current activation curve shifted to more negative potentials 11.41mV, while little effect on the inactivation curve.Activation curve to the right to postpone the activation process makes the current decrease; inactivation inactivation curve to the left so early, the current also decreases, which is stronger than the inhibitory effect of lanthanum ions is an important cause of ytterbium ions.

Whole-cell patch-clamp study Ⅱ: (1) MC3T3 cell outward potassium current with the calcium-dependent, and their half of the increased concentration of EC50 for the 39.8nmol / L.(2) La concentration dependently inhibited MC3T3 potassium currents, the half-inhibitory concentration EC50 value of 8.23μmol / L.(3) of lanthanum on potassium current inhibition kinetic analysis, we found that half of LaCl3 the activation of potassium currents shifted to positive potential voltage of 21mV, so half inactivation voltage shifted to negative potential of 9.66mV.(4) Yb ions on the potassium current and dynamic process have no significant effect.These results suggest that lanthanum and ytterbium on rat osteoblast MC3T3 cell outward potassium current and its role in the dynamics are different.

In short, the paper used MTT, FACS flow cytometry, two rare earth ions La3 +, Yb3 + on NIH3T3 and MC3T3 cell proliferation, cell cycle and apoptosis; using whole cell patch clamp technique, first studied the rare earth ion La3 +, Yb3 + on NIH3T3 and MC3T3 cell size and outward potassium current activation and deactivation kinetics of the effect of the level from the channel in potassium channels clarified the important role of cell growth.Through this study further reveal the biological effects of rare earth and its molecular mechanism of biological effects of in-depth understanding of the mechanisms of rare earth and their potential medicinal value of great significance.