Effect of Sonic hedgehog gene-modified bone marrow mesenchymal stem cells on graft-induced retinal gliosis and retinal ganglion cells survival in diabetic mice

AIM: To investigate the effects of Sonic hedgehog (Shh) gene-modified bone marrow mesenchymal stem cells (MSCs) on graft-induced retinal gliosis and retinal ganglion cells (RGCs) survival in diabetic mice. METHODS: Bone marrow-derived MSCs were genetically modified with the Shh gene to generate a stably transfected cell line of Shh-modified MSCs (MSC-Shh). Intravitreal injections of MSC-Shh and green fluorescent protein-modified MSCs (MSC-Gfp; control) were administered in diabetic mice. After 4wk, the effects of MSC-Shh on retinal gliosis were evaluated using fundus photography, and markers of gliosis were examined by immunofluorescence and Western blotting. The neurotrophic factors expression and RGCs survival in the host retina were evaluated using Western blotting and immunofluorescence. The mechanisms underlying the effects of MSC-Shh was investigated. RESULTS: A significant reduction of proliferative vitreoretinopathy (PVR) was observed after intravitreal injection of MSC-Shh compared to MSC-Gfp. Significant downregulation of glial fibrillary acidic protein (GFAP) was demonstrated in the host retina after MSC-Shh administration compared to MSC-Gfp. The extracellular signal-regulated kinase 1/2 (ERK1/2), protein kinase B (AKT) and phosphatidylin-ositol-3-kinase (PI3K) pathways were significantly downregulated after MSC-Shh administration compared to MSC-Gfp. Brain-derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) levels were significantly increased in the host retina, and RGCs loss was significantly prevented after MSC-Shh administration. CONCLUSION: MSC-Shh administration reduces graft-induced reactive gliosis following intravitreal injection in diabetic mice. The ERK1/2, AKT and PI3K pathways are involved in this process. MSC-Shh also increases the levels of neurotrophic factors in the host retina and promoted RGCs survival in diabetic mice.     

基于Sonic Hedgehog通路探讨复方胃炎合剂干预慢性萎缩性胃炎癌前病变的机制

目的 观察复方胃炎合剂对慢性萎缩性胃炎癌前病变(precancerous lesions of gastric cancer,PLGC)脾虚痰湿热瘀证大鼠Sonic Hedgehog(Shh)通路相关基因表达的影响,并探讨其抑制PLGC进展的机制.方法 SPF级雄性Wistar大鼠随机分为空白组、空白+中药组、模型组、...     

肾衰灵方作用于SHH信号通路对UUO大鼠肾间质纤维化的影响＊

目的 探讨当代名医国医大师郑新经验方——肾衰灵方对单侧输尿管梗阻（UUO）大鼠肾间质纤维化的抑制作用。方法 1. 50只雄性SD大鼠随机分为五组：假手术组、模型组、肾衰灵方低剂量组、肾衰灵方中剂量组、肾衰灵方高剂量组。除假手术组，其余各组结扎单侧输尿管建立肾间质纤维化动物模型，肾衰灵方低中高剂量组予肾衰灵方（10、20、30 mg·kg^-1）连续灌胃14天。检测各组大鼠血清肌酐（SCr）、尿素氮（BUN）、谷丙转氨酶（ALT），谷草转氨酶（AST），碱性磷酸酶（ALP），乳酸脱氢酶（LDH）。采用real-time PCR检测结缔生长因子(connective tissue growth factor，CTGF)核酸水平，采用Masson染色方法检测肾组织观察肾小管和间质病理形态变化。2. 根据结果选择肾衰灵中剂量组为治疗组，再将30只雄性SD大鼠分为假手术组、模型组、肾衰灵方治疗组共3组。用Western blot方法检测音猬因子（Sonic hedgehog，SHH）、神经胶质瘤相关癌基因同源蛋白-1（Glioma related cancer gene homologous proteins 1，Gli1）、锌指转录因子（Snail Family Zinc Finger 1，Snail）蛋白的表达。结果 1.与模型组相比，模型组和肾衰灵各剂量组大鼠血清SCr和BUN的表达水平均明显下调，其中肾衰灵中剂量组疗效最佳（P < 0.05）；Masson染色发现相较模型组，肾衰灵方各剂量组肾间质纤维化明显减轻，其中肾衰灵中剂量组疗效最佳（P < 0.05）；real-time PCR结果发现肾衰灵各剂量组CTGF核酸水平均较模型组下调，其中肾衰灵中剂量组核酸水平最低（P < 0.05）。 2. 相较假手术组，模型组与肾衰灵方组SHH、Gli1、Snai1蛋白的表达均升高（P < 0.05）；与模型组相比，肾衰灵方组SHH、Gli1、Snai1蛋白水平明显降低（P < 0.05）。结论 单侧输尿管梗阻（UUO）大鼠模型中，肾衰灵方可以通过抑制肾间质纤维化改善大鼠肾功能，其作用可能与其阻断SHH信号通路相关。