| Objective： Huntington’s disease (HD) is an autosomal dominant genetic diseasecharacterized by degeneration, so far there is no effective treatment method.Human amniotic epithelial cells (hAECs) possess stem cell-like properties, and exertimmune regulation, thus it is possible for treatment of neurodegenerative disease.Whether they can repair established HD model is unknow. In this study, we aimed toinvestigate the theraperentic effect of hAECs on rats with experimental Huntington’sdisease and its mechanism.Method：①hAECs were isolated from amniotic tissue with mechanical methods andtrypsin digestion method，then phenotype of the passage3（P3）hAECs was identifiedby flow cytometry (FCM) and immunocytochemistry（IC）.②HD rat model induced byinjection of quinoline acid(QA) in the right striatum of SD rats. Model animals weredivided into three groups randomly: model group(n=16), medium control group(n=16),and hAECs group (n=16). In addition to, plus normal group(n=16) as parallel control.hAECs groups injected10μl suspension of hAECs (6×105cells) into damaged striatum,medium control group injected equivalent serum-free L-DMEM medium into the sameway.③At4weeks and8weeks after transplantation of hAECs，changes of motor andrecongnition of rats from several groups were observed by rotation induced withsubcutaneous injection of apomorphine and Morris water maze experiment.④Thepathological change in the striatum was observed by HE staining.⑤The survival timeand differentiation of transplanted hAECs in damaged striatum were observed byimmunofluorescence, The microglia proliferation and GABA neuron expression in thestriatum were detected respectively by immunohistochemical and immunnofluorescencestaining.⑥Percentage of Th1、Th2、Th17、Treg、Tc1and Tc2lymphocyte subsets inperipheral blood mononuclear cells were analysed by FCM at4weeks and8weeks aftertransplantation of hAECs.Results：①Caltured P3hAECs highly expressed CD29,CD44,CD73and were positive for CK19，did not express CD34,CD45,CD71,CD86and HLA-DR.②Comparing withmodel group and medium control group respectively, the rotational frequency of hAECstreated rats induced with apomorphine were reduced at4week and8weeks aftertransplantation(all P<0.05).③Inflammatory respouse in the straitum injuried with QAwas reduce obriously in the hAECs treated rats.④The proliferation of microglia (CD68positive) in the striatum of HD rats was decreased significantly in the hAECs group,while expression of GABA neurons (DARPP-32positive) was increased remarkably.⑤Immunofluorescence staining results showed that the implantated hAECs survived indamaged striatum for8weeks and express neurons nucleoprotein.⑥Comparing withnormal group, the percentage of Treg cells in model group were decreased obviously（P＜0.05）,while the percentage of Treg in hAECs group were higher than medium controlgroup and model group respectively（all P＜0.05）.Conclusion： The hAECs can markedly improve motor fuction and straitumneurodegenerative injury of rats with HD, and its mechanism may relate to inhibitinflammation respouse and microglia proliferation, protect GABA neurons, as well asup-regulate Treg cells. These data suggest that hAECs may be a reliable source of donorcells for cellular therapy of HD. |
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