化瘀散结片防治实验性增生性玻璃体视网膜病变的作用及机理研究
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摘要
目的:研究化瘀散结片对实验性增生性玻璃体视网膜病变(PVR)的防治作用,探
    讨其作用机理。
     方法:采用眼穿通伤+注血法制备PVR模型,进行眼底及常规组织病理学观察,
    ELSA法检测玻璃体中血小板源性生长因子(PDGF)的浓度,用免疫组织化学方
    法观察增生膜与血管内皮纽胞表面血管细胞粘附分子-1(VCAM-1)及细胞间粘附
    分子-I(ICAM-1)的表达。同时,采用体外细胞培养技术,培养兔视网膜色素上皮
    细胞,井结合中药血清药理学研究方法,进行化瘀散结片防治PVR的体外实验研
    究。
    结果:体内及体外实验均证明化瘀散结片有防抬PVR的作用。(1)眼底及常规病
    理切片观察结果显示,化瘀散结片高、低剂量治疗组增生膜形成较模型对照组缓慢,
    且增生程度较轻,增生膜面积及周长与模型对照组相比显著减小(P〈0.01).各用药
    组间比较,低剂量治疗组增生膜面积明显大于高剂量治疗疗组(P〈0.05),说明化瘀散
    结片的抗增生作用存在一定量效差异。化瘀散结片高低剂量治疗组的作用与阳性对
    照组比较有极显著性差异(P〈0.01).(2)化瘀散结片高低剂量治疗组视网膜神经节
    细胞数量,均明显高于模型对照组(P〈0.05或P〈0.01). 化瘀散结片高低剂量
    治疗组玻璃体腔内PDGF浓度明显低于模型对照组(P〈0.01),阳性对照组也明显低
    于高低剂量治疗组(P〈0.05)及(P〈0.01).化瘀散结片能明显降低ICAM-l、
    VCAM-1在增生膜及血管内皮细胞表面的表达,与模型对照组比较有极显著或显著
    差异(P〈0.05)或(P〈0.01),各用药组之间比较则无明显差异(P〉O.05).(5)高剂
    量的化瘀散结片含药血清对体外培养的视网膜色素上皮细胞抑制作用最强,(抑制
    率为53.52%),而道诺霉素(80ng/ml)与低剂量化瘀散结片含药血清的抑制率分别
    为41.31%及 38.95%。
    结论:化瘀散结片能有效防止PVR的形成与发展,其作用机理与减少玻璃体中血
    小板源性生长因子(PDGF)浓度,降低视网膜增生膜上和血管内皮细胞表面
    ICAM-I、VCAM-1的表达,以及直接抑制体外培养的视网膜色素上皮细胞增生有
    关.
BACKGROUND: The Huayusanjie Tablet is used as a therapeutic option for animals with experimental PVR . The aim of this study was to investigate the effect and mechanism of Huayusanjie Tablet on the experimental PVR in vitro and in vivo. METHODS: Autologous whole blood was injected into the vitreous cavity of albino rabbits after canalization,thus induced experimental PVR model. Concentration of PDGF .in vitreous cavity was measured by ELISA method, and the expression of ICAM-1 and VCAM-1 was observed by immunohistochemicai examination. RPE proliferation was measured by MTT assay.
    RESULTS: There was preventive and curative effect of Huayusanjie Tablet on. experimental PVR in vitro and in vivo. (1) Result of fundus and pathology-microscope examination showed the average stage of PVR in the eyes treated with Huayusanjie Tablet (9.6g/Kg or 2.4 g/Kg) was significantly lower than that in the control group on days 28(P<0.01),and there was significant difference between the high-dose group and low-dose group(p<0.05).However , the effect of Huayusanjie Tablet was obviously lower than daumomicin(P<0.01). (2) Huayusanjie Tablet could increase the number of ganglion cell in retina(P<0.05 or P<0.01). (3) The concentration of PDGF in the vitreous of Huayusanjie Tablet -treated eyes was significantly lower than in the control group. And daumomicin was also lower than Chinese medicine. (4) Expression of ICAM-1 and VCAM-1 were observed on proliferative membranes and vascular endothelial cell. Huayusanjie Tablet could decrease the expression of two cellular adhesion molecule described above, but there was no difference within Huayusanjie Tablet groups and daunomicin group. (5) High-dose of Huayusanjie Tablet could inhibit proliferation of cultured RPE cell in vitro,while low-dose group and daunomicin group(80ng/ml) showed lower inhibitory effect on RPE.
    CONCLUSION: The data suggest that Huayusanjie Tablet can prevent the formation and development of PVR in' vitro- and in.vivo.Its mechanism concern with factors described below:lowering the concentration of PDGF,decreasing the expression of ICAM-1 and VCAM-1 in proliferative membrane and vascular endothelial cefl,directly inhibiting proliferation of RPE. in vitro.
引文
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