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In vitro suitable thermal stimulus collaterals related vascular endothelial cells after the start of "Tongluo blood" effect of cell biological mechanisms

Author: WuLi
Tutor: YangXiaoFang
School: Guiyang College of Traditional Chinese Medicine
Course: Acupuncture and Massage
Keywords: collaterals moxibustion suitable for thermal stimulation umbilical vein endothelial cells endothelin-1(ET-1) thrombomodulinprotein (TM) prostacycl in (PGI2)
CLC: R245
Type: Master's thesis
Year: 2013
Downloads: 11
Quote: 0
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Abstract


Objective:Take the venae related vascular endothelial cells as research carrier, to observe changes of contents that vascular endothelial cells secrete vasomotor regulation substances ET-1, TM, PGI2content and further explore basic cell biological mechanisms how to start after receiving stimulations under the essence of moxibustion appropriate warm stimulation.Methods:Using intravenous infusion enzyme digestion in vitro to culture human umbilical vein endothelial cells and applying immunocytochemical to identify umbilical vein endothelial cells with (Ⅷ factor). Umbilical vein endothelial cells were divided into Control group (37℃group) and Suitable thermal stimulation group (40°C group), suitable for the warm stimulation group40°C stimulus30min,2times a day,3days of continuous stimulation. The inverted phase dynamic changes in cell morphology were observed under the microscope. Detected by ELISA (enzyme linked immunosorbent assay) method with the vascular endothelial cells in vitro suitable warmed stimulation the release of ET-1and TM, content of the EIA (enzyme immunoassay) method detection in vitro suitable warmed under the stimulation of vascular endothelial cells release PGI2content.Results:1. Changes in cell morphology:primary cultured human umbilical vein endothelial cells (Human umbilical venous endothelial cells, HUVECs) morphology is small polygonal, short spindle or spherical cell monolayer, and non-overlapping. HUVECs fusion flat polygons, often isolated and the typical paving stones or pebbles like arrangement, clear boundary, clear nucleus, the nucleus is oval or round, the chromatin in the nucleus is sparse, empty and bright nucleolar1-2a. Passaged HUVECs showed mostly typical paving stone or pebble-like arrangement, clear boundary clear nucleus, the nucleus is oval or round, the chromatin in the nucleus is sparse, empty and bright, nucleolar1-2; transmitted to3-4generation HUVECs, the growth rate slowed, refraction increase, increasing the cell body, cell mutation fine particles and vacuoles in the cytoplasm, mitotic reduce a series of signs of aging. Appropriate thermal stimulation, HUVECs morphological changes are not obvious, a few cells contour round cell gap increases, cobblestone-like arrangement, clear boundary, clear nucleus, the nucleus is oval or round, sparse chromatin in the cell nucleus and empty light. HUVECs with DAB immunocytochemistry Ⅵ factor staining, after finalizing the light microscope to brown granules in the cytoplasm.2. Vascular endothelial cells of ET-1content of40°C is lower than37℃thermal stimulation, and the difference was significant (P<0.05); the TM content in40℃is higher than37℃group, and the difference is highly significant (P<0.01); PGI2content in40℃is higher than37°C group, and the difference was significant (P<0.01).Conclusions:1. Appropriate thermal stimulation can relax vessels through reducing the content of ET-1in vascular endothelial cells, which is the way to realize the role of how to dredge collateral and promote the circulation of blood.2. Suitable thermal stimulation can promote the content of TM in vascular endothelial cells, which may maintain the normal circulation of blood through anticoagulation so as to achieve the efficacy of promoting blood Circulation of collaterals.3. Suitable thermal stimulation can elevate the release of PGI2in vascular endothelial cells, which may dredge collaterals through promoting the relaxations of vessels on one hand and may promote the circulation of blood through inhibiting platelet aggregation.4. Suitable thermal stimulation in vitro, the efficacy of dredging collaterals and promoting the blood circulation may result from promoting the release of TM, PGI2and inhibiting secretion of ET-1. And it realize the efficacy by means of maintaining the normal blood circulation and systolic and diastolic function.

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