ABSTRACT
Systemic sclerosis (SSc) is a chronic autoimmune disease that can involve multiple organs throughout the body. It is characterized by extensive vascular disease and fibrosis of the skin and internal organs, but its mechanism is still unclear. Studies have confirmed that the Wnt pathway is involved in SSc fibrosis, but its pathological role in vascular lesions has not been reported. In this study, the bleomycin (BLM)-induced SSc mouse model was used to investigate the role of Wnt pathway in SSc cutaneous vascular lesions. Eighteen BALB/C mice were randomly divided into control, model and treatment groups. The control group of mice was injected with PBS 100 μL/d. The model group was injected with BLM 100 μL/d at a concentration of 1 mg/mL. The treatment group of mice received injection of BLM 100 μL as the model group of mice and retroperitoneal injection of iCRT3 (Wnt and beta-catenin inhibitors) 5 mg/kg/d. Mice were sacrificed on day 28. The thickness of dorsal dermis and epidermis in the model group induced by BLM was significantly higher than that in the control group (P <0. 05). The skin appendages such as sebaceous glands and hair follicles in the model group were significantly reduced. At the same time, the thickness of fat layer became thinner and surrounded by fibrous tissue, and the collagen deposition in the model group was higher than that in the control group. It was identified at the histological level by immunohistochemical staining that α-SMA expression in model group and treatment group α-SMA is highly expressed in skin tissues, and the positive expression of α-SMA around blood vessels was significantly higher than that in the control group. In addition, the expression of IL-6 and IL-17 in serum of model group was significantly higher than that in control group (P<0. 05), and the expression of IL-6 and IL-17 in serum of treatment group was significantly lower than that in model group (P<0. 05). Furthermore, q-PCR analysis showed that the mRNA levels of β-catenin in the skin microvessels of the mice were higher in the model and the treatment groups as compared with the control group. The protein levels of Wnt5A, β-catenin, α-SMA and col1A1 were analyzed by Western blotting and results showed that the levels of fibrosis-related proteins, α-SMA and col1A1, were increased in the model group injected with BLM as compared with the control group (P<0. 05), whereas iCRT3 treatment attenuated the upregulation of α-SMA and col1A1 induced by BLM in the treatment group (P<0. 05). The levels of Wnt pathway-related proteins β-catenin and Wnt5A were significantly increased in the model group as compared with the control (P<0. 05). This study suggests that BLM can successfully induce the skin phenotype of mice with systemic sclerosis. Abnormal activation of Wnt signaling pathway is involved in BLM-induced skin microangiopathy in mice with scleroderma, and the specific Wnt pathway inhibitor iCRT3 can reduce BLM-induced scleroderma. The expression of α-SMA and col1A1 proteins in mouse skin microvessels can improve the microvascular lesions of mouse skin. Inhibition of Wnt pathway may directly or indirectly down-regulate the abnormal expression of cytokines IL-6 and IL-17, and interfere with BLM-induced progression of vascular lesions in mice.
ABSTRACT
Objective:To explore the mechanism of the embolism and sclerotherapy of fibrin glue combined with bleomycin (FG/BLM) for the treatment of cervicofacial vascular malformations by color doppler ultrasound.Methods:10 patients with venous malformation(VM) and 10 patients with arterio-venous malformation(AVM) were included.All patients underwent embolism and sclerotherapy of FG/BLM guided by ultrasound.Color doppler ultrasound was used to record the real-time two-dimensional ultrasonography and color doppler image.The flow and distribution of FG/BLM after injection into the lesions were observed.Results:Two-dimensional ultrasonography showed clumps or flake strong echo after immediate injection of FG/BLM into the cavity of VMs,then floated in the abnormal venous lumen and diffused throughout the cavity.At the later stage the lesions were filled by a large number of flocculent and netted low echo,and patchy strong echo.The volume of VMs cavity expanded dramaticlly,and the blood flow signal was significantly decreased.After injection of FG/BLM into the lumen of AVMs,clumps or flake strong echo were observed,then most of the snowflake strong echo rapidly filled or scattered along with blood stream to the distal part of the vessels.The color doppler showed significantly decrease of blood flow signal.Conclusion:FG/BLM injection can embolize and block the draining vein of VM,and play a role on the storage of sclerozing agent.FG/BLM injection can embolize both the dilated blood vessels and capillary network of AVM.
ABSTRACT
Objective:To establish a multidrug resistant human salivary gland adenoid cystic carcinoma cell line.Methods:Human salivary gland adenoid cystic carcinoma cell line ACC-2 was treated by 24-hour-exposure to high dose of Bleomycin(BLM)(20 ?g/ml).Drug sensitivity was evaluated by MTT assay.Cell counting was employed to make the growth curve and to calculate the cell doubling time.Flow cytometry(FCM) was used to study the cell cycle distribution and apoptosis.The colony formation ability was also observed.Results:Multidrug resistant cell line of human salivary gland adenoid cystic carcinoma was established and named ACC-2/BLM.After 10 times repeated exposure to BLM,the resistance index(RI) to BLM,5-Fluorouracil(5-FU),Cisplatin(CDDP),Cyclophosphamide(CTX),Vincristine(VCR) were 7.299,1.03,2.15,1.114 and 5.96 respectively.Compared with ACC-2,the proliferation potential of ACC-2/BLM cells was decreased.The ACC-2 apoptosis cells were much more than ACC-2/BLM cells after 9-day-treatment by BLM at 60 ?g/ml.Conclusion:ACC-2/BLM cell line has multidrug resistant characteristics.