Therapeutic effects of transplantation of lentivirus-mediated artemin gene modified bone marrow mesenchymal stem cells on the rat model of Parkinson's disease

Abstract

Objective To investigate the potential therapeutic efficacy of lentivirus-mediated artemin (ARTN) gene modified bone marrow mesenchymal stem cells (MSCs) transplantation on the rat model of Parkinson's disease (PD) and the effects on expression of brain-related proteins. Methods MSCs were isolated and cultured in vitro, transfected by recombinant lentiviral vectors carrying ARTN gene. The PD rat model established by 6-hydroxydopamine (6-OHDA) was randomly divided into 5 groups: Sham group, PD group, MSCs group, MSCs transfected with empty lentiviral vectors transplanted (LV-MSCs) group and MSCs transfected with recombinant lentiviral vectors carrying ARTN gene transplanted (LV-ARTN-MSCs) group. The MSCs, LV-MSCs and LV-ARTN-MSCs groups were transplanted into the left striatum of each rat model of PD and ethology tests in every group were made with intraperitoneal injection of apomorphine (APO) 2, 4, 6, 8 weeks after transplantation. The expression of tyrosine hydroxylase (TH) protein in substantia nigra (SN) was measured by Western blotting and immunohistochemistry, and immunofluorescence showed ARTN gene modified MSCs expression in rat brain tissue. The levels of dopamine (DA), dihydroxy-phenylacetic acid and homovanillic acid in striatum of each group were detected by high performance liquid chromatography. Results After injection of APO, rotation frequency decreased in LV-ARTN-MSCs group, i. e. (179.33±10.74) circles/30 min vs (235.83±18.95), (203.67±11.50) and (206.33±11.86) circles/30 min in PD, MSCs and LV-MSCs groups (q=8.828, P<0.01; q=3.802, P<0.05; q=4.219, P<0.05). The percentage of TH-positive cells in SN after cell transplantation was increased significantly in LV-ARTN-MSCs group (64.05%±5.49%) when compared with PD group (34.18%±3.35%), MSCs group (52.59%±4.48%) and LV-MSCs group (50.57%±4.41%), respectively (q=13.280, 5.135, 6.028, all P<0.01). At the same time, TH protein in SN after cell transplantation was also increased obviously in LV-ARTN-MSCs group. ARTN gene modified MSCs can survive for at least 6 weeks in the rat brain of PD, mainly concentrated in the transplantation side of striatum. Eight weeks later, the levels of DA in striatum after cell transplantation were elevated significantly in MSCs group (2.34±0.54), LV-MSCs group (2.28±0.45) and LV-ARTN-MSCs group (2.28±0.45) when compared with PD group (0.87±0.07) (q=5.233, P<0.05; q=5.020, P<0.01; q=20.190, P<0.01), and LV-ARTN-MSCs group showed the most significant improvement. Conclusion ARTN gene modified bone marrow MSCs transplanted into the striatum of brain may have therapeutic effects on rat models of PD. Key words: Parkinson's disease; Mesenchymal stem cells; Nerve tissue proteins; Lentivirus; Transfection; Disease models, animal