| Background and Objective: Thrombocytopenia is a common clinicalsyndrome as a side effect in treatment such as high dosage chemotherapy, radiotherapy andstem cell transplantation, It become one of main causes of death. Nowadays, plateletstransfusion is an effective and extensively used way to retrieve the thrombocytopenia. Butthat can induce the anti-platelets antibodies and invalidate the platelet transfusion ifreduplicative applicated. Recent years, with the discovery and purification ofthrombopoietin (TPO) and recombination human interleukin-11 (rhIL-11),thrombocytopenia was partly cured, but some problems still existed. For example, the TPOcan induce the antibodies and activate the platelets to form the thrombus. And the rhIL-11also has some shortcomings such as that it need more time to take effective. And it is veryexpensive. These disadvantages restrict the use of TPO and rhIL-11. How to promote plateletcounts quickly and avoid antibodies and other adverse reaction has become an importantclinical problem.Therefore, it is very important in clinic to find some materials to replace TPO or toimprove its effect. That can stimulate the cell proliferation and differentiation ofmegakaryocyte. And then eventually increase the platelet number of peripheral blood. Theform of platelet is on the basement of megakaryoblasts. We can draw assistance from somematerial to find a new drug or ways to stimulate the megakaryoblasts. We can prove howand why the material can produce a marked effect. And then we can master the characteristic ofmegakaryoblasts. That is very useful. The material needs to have some functions that it cansimulate the cells proliferation and differentiation of MK directly. Or it can simulate theproliferation and differentiation of MK indirectly, by influencing mesenchymal stem cellsin the bone. Or it can directly and also indirectly to simulate the proliferation anddifferentiation of MK.From the work we have done, we have discovered that 5-HT can stimulate themegakaryoblasts. And it also can anti-apoptosis of the megakaryocyte. That shows that the 5-HT can stimulate the cells proliferation of MK. And it needs to be proved whether it also can stimulate thecells proliferation of MK by influencing mesenchymal stem cells in the bone marrow.We are ready to observe the cells proliferation of 5-HT to the bone marrowmesenchymal stem cells. And we will observe the quantity of interleukin 11(IL-11) andinterleukin 6(IL-6) secreted by mesenchymal stem cells when 5-HT was added. We want toconfirm whether the 5-HT can promote the proliferation of MSC as well as enhance thesecretion of IL-11 and IL-6, therefore may stimulate the megakaryocyte proliferation.Methods: The bone marrow is drawn from the BABL/c mice. The bone marrowmesenchymal stem cells can be isolated by using the percoll liquid. The zero and the firstparental generations of the MSC are cultured with blood serum in vitro. The second parentalgeneration is cultivanted without blood serum for 24 hours. Then cells are divided into four groups: the5-HT 100nmol/L, the 5-HT 200nmol/L, the 5-HT 400nmol/L and the control group which without 5-HT.Microscope was used to observe the cells proliferation of MSC which are influenced by differentconcentrations of 5-HT on 0 hour, 6 hours, 12 hours, 24 hours, 48 hours. And we also use the microscopeto observe the cell proliferation of MSC which from control group. We use the confocol laser scanningmicroscopy to detect the change of IL-11 and IL-6 on the cells from the different four groups on 0 hour,6 hours, 12 hours, 24 hours and 48 hours. And we also use the ELISA to measure the change of IL-11and IL-6 in supernatant from the four groups on 0 hour, 6 hours, 12 hours, 24 hours, and 48 hours.Results: We find that all groups with different concentrations of 5-HT can stimulate theproliferation of MSC, and that with 200nmol/L has the most significant effect. Meanwhile, the result ofthe cells proliferation from the 24 hours is also the most significant (P＜0.01) when comparing toother different time. The results of the IL-11 and IL-6 which getting from the confocol laser scanningmicroscopy prove that 200nmol/L has the most significant effect to stimulate the secretion of IL-11 andIL-6 when at 6 hour, 12 hour, 24 and 48 hour. The changes of 400nmol/L is no statistics significance(P＞0.05) comparing to the group of 200nmol/L. The different hours' results of the comparegroup are also having no significance (P＞0.05). There is significance in 6 hours and 12hours when comparing to the 0 hour. And there is statistics significance in 24 hours whencomparing to the 0 hour. And there is no significance in 24 hours when comparing to the 48.Using the ELISA, we find the results are similar to those results which gotten from theconfocal laser scanning microscopy. Conclusions: All the results show that 5-HT has the functions to stimulate thecells proliferation of MSC, as well as stimulate the secretion of the interleukin-11 andinterleukin-6.
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