Efficacy And Experimental Study Of Mechanism Of Propranolol Treatment For Infantile Hemangioma

OUTLINEHemangiomas are the most common congenital benign tumor of infants, occurring in2%-3%of neonates and10%of children<1year of age. Propranolol, a nonselective (3-adrenergic receptor blocker commonly used for arrhythmia, angina pectoris, hypertension. In2008, Leaute-Labreze et al observed that propranolol can control the growth of IHs effectively, especially the proliferation of severe hemangiomas. After that, many groups worldwide have initiated propranolol therapy IHs and it was confirmed that propranolol has significant effective for IHs.A rapid halt in lesion progression and evidence of regression were confirmed in the majority of the cases. Propranolol has become the first-line treatment drug for IHs replaced the hormone. However, there are still a lot of potential side effects, especially in the early of oral propranolol,and required hospitalization to detect heart rate, blood pressure, blood sugar and so on. It has many inconvenience for parents and physicians. In view of this, in chapter Ⅰ-part I:A retrospective chart review was performed on25children (21female and4male) with a median age of4months (range,1-10months). A total of28lesions were treated with1%propranolol ointment. It was showed that Topical therapy with1%propranolol ointment may be a safe and effective method for the treatment of superficial IHs and can be used as an adjuvant treatment measure during the wait-and-see period. In addition, it is currently in dispute that the dosage of propranolol used and the duration of treatment. In chapter Ⅰ-part Ⅱ:A retrospective review (106cases)was performed of IH treated with propranolol at the department of aesthetic, plastic, and burn surgery of the provincial hospital affiliated with Shandong University, Jinan, China, from Sept.16,2009to June1,2013. to describe the dosage of propranolol used, the duration of treatment, the effects and side effects of propranolol treatment. Propranolol administered orally at1to1.5mg/kg/day had a rapid therapeutic effect with few complications.It was unknown that the mechanism of abnormal proliferation on hemangioma. Folkman believed that it was a angiogenesis disease, and the major factor was an uncontrolled angiogenesis in proliferation of hemangioma. The process includes endothelial cell proliferation, extracellular matrix degradation selectivity, increased vascular permeability and migration of cells. The cell growth factor is a class of substances which induce or promote the formation of new blood vessels, regulate endothelial cell proliferation, and have a key role in the ndevelopment of hemangiomas. EGFL7, located in human9q human chromosome, was a chemoattractant for endothelial cellsand is up-regulated in angiogenesis. In chapter Ⅱ-part Ⅱ:The expression of EGFL-7was detected by immuno-histochemical technique in cases of proliferating hemangiomas, involuting hemangiomas, involuted hemangiomas, and normal skins. To detect the expression of EGF-like domain-containing protein-7(EGFL-7) in the different stages of infantile hemangioma, approach the effect and significance of EGFL-7in the occurrence and development of infantile hemangioma. In chapter Ⅱ-part II:Under propranolol intervention, it was observed human umbilical vein endothelial cells (HUVEC) proliferation, apoptosis, lumen formation and expression, EGFL7, to explore the mechanism of inhibition of propranolol hemangioma hyperplasia.Chaper Ⅰ. The efficacy of propranolol treatment of infantile hemangiomaPart Ⅰ. Topical propranolol for treatment of superficial infantile hemangiomasOBJECTIVE:The aim of this study was to evaluate the efficacy and safety of1%propranolol ointment in thetreatment of superficial IHs.METHODS:A retrospective chart review was performed on25children (21female and4male) with a median age of4months (range,1-10months). A total of28superficial IHs were treated with1%propranolol ointment. Topical propranolol was applied thrice daily for a mean duration of21weeks (range,5-59weeks). Changes in the size, texture, and color of the tumor were monitored and recorded at regular intervals. The treatment response was evaluated using a3-point scale system: good, partial, and no response. Adverse effects after medication were evaluated and managed accordingly.RESULTS:Of the28hemangiomas,16(57%) demonstrated good response,17(34.7%) showed a partial response, and3(10%) had no response. Among all the IHs,90%(95%confidence interval72%-98%) showed either good or partial responses to topical1%propranolol ointment treatment. No systemic complication was observed in any of the patients.CONCLUSION:Topical therapy with1%propranolol ointment may be a safe and effective method for the treatment of superficial IHs and can be used as an adjuvant treatment measure during the wait-and-see period. Part Ⅱ Oral propranolol in the management of infantile hemangiomaOBJECTIVE:to describe the side effects of propranolol treatment in infantile hemangiomas(IHs).METHODS:A retrospective chart review was conducted on all children who attended clinic from Sept.16,2009to Nov.11,2013.106children (71girels and35boys, mean age and weight at onset of treatment:5.1months and7.3kg). Prorpanolol was administered in a progressive schedule to1.0to1.5mg/kg per day, divided in three doses. Demographic data, clinical features, imaging findings treatment regimens of propranolol, and outcome were investigated. Adverse effects after medication were evaluated and managed accordingly.RESULTS:All106patients had a good response. Side effects that needed intervention and/or close monitoring were included diarrhea (n=10), hypotension (n=7), and milk regurgitation (n=2). Nightmares (n=2), agitation (n=1), and cold extremities (n=1) were observed.CONCLUSIONS:Propranolol administered orally at1tol.5mg/kg/day had a rapid therapeutic effect with few complications. Chaper Ⅱ. The experimental study of propranolol treatment of infantile hemangiomaPart Ⅰ. The expression and significance of EGFL-7in infantile hemangiomaOBJECTIVE:To detect the expression of EGF-like domain-containing protein-7(EGFL-7) in the different stages of infantile hemangioma, approach the effect and significance of EGFL-7in the occurrence and development of infantile hemangioma.METHODS:Collect infantile hemangioma specimens embedded in paraffin, the specimens were all excised by operations and were confirmed by pathologic diagnosis. After routinely HE stain, we classified the specimens according to the Mulliken’s classification standard, including proliferating group, involuting group and involuted group, normal skin tissues were chosen as control group. The expression of EGFL-7was detected by immuno-histochemical technique in cases of proliferating hemangiomas, involuting hemangiomas, involuted hemangiomas, and normal skins; the mean optical density was measured by image analysis system.RESULTS:Positive expression of EGFL-7was located in cytoplasm tumor cells and nomal skin tissues. The strongly positive or positive expression in proliferating phase, positive expression in involuting phase, and weakly positive or negative expression in involuted phase and normal skin tissuesCONCLUSION:EGFL-7are involved in the occurrence and development of infantile hemangioma and there is a close relationship between them. Part Ⅱ. The effect of propranolol in biological behavior of and expression of EGFL-7of HUVECOBJECTIVE:To investigate effects of propranolol in biological behavior and EGFL-7of the human umbilical vein endothelial cells in vitro. It would be provided a theoretical basis for further study of propranolol for infantile hemangiomas mechanism.METHODS:The human unbilical vein endothelial cell line was cultered by propranolol with varying concentration. Cell growth was assessed by MTT, the apoptotic rate was assessed by Annexin V-FITC/PI double staining method, the tube formation ability was assessed by tube formation assay. The expression of EGFL-7mRNA and protein were identified by semi-quantitive reverse transcriptase polymerase chain reaction(RT-PCR) and western blot, respectively.RESULTS:After drug processing for24h and48h, compared with the normal control group, the inhibition rates of25μmol/L,50μmol/L,100μmol/L,125μmol/L propranolol on proliferation of HUVEC were found an upward trend (P<0.01) and then we could acquire suitable concentration by calculated IC50=125.33±2.71μm. After cultered for24h, the early apoptotic rates of Oμmol/L,25μmol/L,125μmol/L were detected2.7665±0.1528(%),15.467±2.3861(%),26.133±1.6503(%)(P<0.05). The tube formation capacity were not significant differences after drug processing (Oμmol/L,25μmol/L,125μmol/L) for24hours, and after48h, it was found that the differences of tube formation capacity. Propranolol can down-regulate the expression of EGFL-7mRNA and protein in a concentration-dependent manner(P<0.05).CONCLUSION:Propranolol inhibited HUVEC proliferation and promoted the apoptosis in a concentration dependent manner, and inhibited the tube formation of HUVEC in a time-dependent manner. EGFL-7are involved in the occurrence and development of infantile hemangioma and there is a close relationship between them.