The Research Of The Influence Of Ajuga Decumbens Thunb On The Growth Of Hemangioma And The Expression Of Fas/FasL In Nude Mice Model

Objective: In order to observe the influence of Ajuga decumbensthunb on the growth of hemangioma and the express of Fas/FasL in nude micemodel, and discuss the effect and the mechanism of Ajuga decumbens thunb asa treatment for hemangioma. Methods: Infant proliferative hemangioma wastaken by surgery and separated into small pieces (3mm*4mm*5mm) understerile condition in operating department. Then, tissue pieces were implantedsubcutaneously into specific pathogen free nude mice, each mouse wasimplanted two tumor tissue pieces.30day after implantation, we had65nudemice that hemangioma survived and grew normally. Randomly selected5to bemeasured the volume of hemangioma and then sacrificed, the hemangiomatissues were taken out and made into paraffin sections, as primary contrast；remaining60nude mice were random separated into three experimental groupsto do randomized controlled trails：vaseline group, Ajuga decumbens thunbgroup and triamcinolone acetonide group, that were measured the volume ofhemangioma and given different medicine. Each group had20nude mice and40tumors. Liniment of vaseline anddimethyl sulfoxide mixture as controlchemistry was administrated every day in vaseline group. Liniment ofself-made Ajuga decumbens thunb was administrated in the surface ofhemangioma every day in Ajuga decumbens thunb group. Triamcinolone acetonide was injected into the hemangioma once at the beginning intriamcinolone acetonide group; the dose of triamcinolone acetonide was0.05ml(4mg/ml). The mice in three groups were fed equally. After be administrateddifferent medicine, the growth and status of tumor was observed. In themeantime, the volume of hemangioma in three groups was measured in vivo.After7day、14day、21day and28day，randomly selected five experimentalmice in each group to be sacrificed, the hemangioma tissues were taken out tobe measured the volume and made into paraffin sections. Hematoxylin-eosin(HE) staining of all paraffin sections and analyzed the histological change.Immunohistochemistry staining of all paraffin sections with Fas、FasL andcysteinyl aspartate specific proteinase8(caspase-8) antibody and analyzed theirexpress. Tested apoptosis by terminal deoxynucleotidyl transferase mediateddUTP nick-end labeling (TUNEL) method, and analyzed their change inhemangioma model after be administrated different medicine. The experimentaldata were analyzed by statistical software SPSS20.0, with P<0.05as there wassignificant difference. Results:1.Before intervention, all hemangiomas inexperimental nude mice grew well. The volume of10tumors in5mice thatrandomly selected to be sacrificed before intervention had no differencecompared with the tumors in three experimental groups(P＞0.05). Observed byHE staining, the appearance of tumors showed there were at proliferative stage.Counted the expression of Fas, FasL and caspase-8by immunohistochemistrystaining， the positive index was28.56±2.43、18.31±1.92、17.48±1.63 respectively. Counted the apoptosis index by TUNEL method, the apoptosisindex was6.79±1.33.2. After intervention, the hemangiomas in vaseline groupgrew continuously and the volume of tumor increased apparently, observed theHE staining paraffin sections by microscopic, the hemangiomas were at theproliferation stage. After be administrated different medicine, compared thevolume of tumors in Ajuga decumbens thunb group and triamcinoloneacetonide group with before, the volume of hemangioma decreased apparently(P<0.05). Observed the HE staining of paraffin sections by microscopic,vascular density reduce, tumor atrophy, degeneration, occlusion of the vessellumen, rupture, fat and fibrous tissues in the vascular infiltration.3. Thepositive index of Fas、FasL and caspase-8in Ajuga decumbens thunb groupwere increased apparently after intervention, and7day after be administrateddifferent medicine, the increase was most apparently; after be administrateddifferent medicine7day,14day,21day, compared with before intervention andvaseline group, triamcinolone acetonide group at the same intervention time,there was significant difference (P<0.05); after be administrated differentmedicine28day, compared with before and vaseline group, triamcinoloneacetonide group at the same administrated time, there was nodifference(P>0.05). After be administrated different medicine, the positiveindex of Fas、FasL and caspase-8in triamcinolone acetonide group and vaselinegroup to be compared with before, there was no difference(P>0.05).4.After beadministrated different medicine, the apoptosis increased apparently in Ajuga decumbens thunb group and triamcinolone acetonide group, the index ofapoptosis to be compared with before and vaseline group at the sameadministrated time, there was significant difference(P<0.05). Conclusion:1.Ajuga decumbens thunb can facilitate the degradation of hemangioma in nudemice model.2. Ajuga decumbens thunb can increase hemangioma tissueexpress Fas, FasL and caspase-8apparently in nude mice model.3. Thetherapeutic effect of Ajuga decumbens thunb on hemangioma may be relate tothe apoptosis mechanism that is activated by the Fas/FasL, and lead toapoptosis of hemangioma in nude mice model.