Effects Of Pingyangmycin Combined With Sodium Hyaluronate On Human Lymphatic Malformation Endothelial Cells

Purpose:Lymphatic malformations(LMs), traditionally called lymphangiomas, are congenital diseases caused by development errors of the lymphatic system. Approximately90%of the patients occur before the age of2years, except a few cases which occur in adulthood, and about75%of the lesions are located in the head and neck region. LMs, though benign, usually do not regress spontaneously. The lesions can grow rapidly with trauma, infection or bleeding, resulting in disfigurement as well as severe impairment of speech, swallow and respiration. The current main treatment of LMs contain surgery, laser therapy and sclerotherapy. All these methods have advantages and disadvantages. We should formulate individualized treatment plan for patients based on specific patient and hospital technical conditions, with comprehensive sequence treatment, to get the best curative effect. In recent years, sclerotherapy as a new treatment for LMs has been gradually developed. sclerotherapy of LMs with Pingyangmycin achieved satisfactory results and compared with surgery and other treatments, sclerotherapy has some advantages, Safe, simple operation, applicable to the wide range of lesion that surgery is not easy to excise completely, without scar and good esthetic effects. The effects of Pingyangmycin sclerotherapy is good for macrocystic lesions but poor for microcystic lesions. In order to improve the efficacy of pingyangmycin, combinations of drugs become a trend for the treatment of LMs. Here, we try to detect if human lymphatic malformation endothelial cells (HLMECs) and human lymphatic malformations (HLMs) tissue express the hyaluronan receptor-1(LYVE1) which is the lymphatic endothelial specific marker and to test the effects of pingyangmycin (PYM) combined with sodium hyaluronate (HA) on HLMECs in vitro in order to provide experimental basis for clinical treatment of LMs with the combination of PYM and HA.Methods:Primary HLMECs were isolated from a HLMs tissue and cultured with endothelial cell medium (ECM). By lymphatic endothelial specific markers LYVE1and PROX1staining, the HLMs specimen and HLMECs were identified. The proliferation effects of the HLMECs treated with gradient densities of PYM (1、10、100、500μg/ml)or HA (50、100、300、500μg/ml) for12h,24h and48h, were compared by MTT assay, and the optimal concentration of PYM and HA was selected. Then,the HLMECs were treated with the selected concentration of PYM and HA for0、12、24、48h together, and the cell proliferation was evaluated by MTT assay. At24h,the apoptosis was evaluated by TUNEL.Results:(1) From the HLMs specimen, we acquired the HLMECs.(2) LYVE1and PROX1were both expressed within the HLMs tissue and these HLMECs.(3)100μg/ml and500μg/ml PYM inhibited cell proliferation obviously. The optimal concentration of PYM was100μg/ml. At24h,300μg/ml HA inhibited cell proliferation mildly. The optimal concentration of HA was300μg/ml.(4) The combination of PYM and HA was more effective than single PYM on inhibiting HLMECs-growth and promoting HLMECs-apoptosis.Conclusions:(1) PYM combined with HA can avoid the defect of free PYM that easy to wash away by the blood flow, can maintain the effective concentration of PYM Within a period of time, extended the action time of PYM on HLMECs;(2) PYM combined with HA can reduce the adverse effects of PYM and was beneficial to the clinical application of PYM;(3) Adsorption of HA to the HLMECs membrane, which not only make the treatment of PYM combined with HA for lymphatic malformation be targeted therapy, but also increase the drug concentration of lymphatic malformation endothelial cells surface, can cause better clinical effects.