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目的观察黑绛丹治疗放射性皮炎的可能作用机制。方法 55只大鼠采用60Co源局部照射建立大鼠Ⅳ度放射性皮炎动物模型,将48只造模成功的大鼠随机分为黑绛丹组、对照1组、对照2组、模型组,每组12只,各组分别外敷黑绛丹溶液、重组人表皮生长因子外用溶液、水溶维生素E、生理盐水1 ml,每日1次,连续给药10天。另设空白组12只,予生理盐水外敷模型组大鼠同样部位。透射电镜观察各组创面组织细胞超微结构,并检测各组创面组织细胞凋亡相关因子,即死亡相关因子(Fas)、死亡相关因子配体(Fas L)、半胱氨酸天冬氨酸蛋白酶3(Caspase-3)、半胱氨酸天冬氨酸蛋白酶8(Caspase-8)、半胱氨酸天冬氨酸蛋白酶9(Caspase-9)、B细胞淋巴瘤2(Bcl-2)、B细胞淋巴瘤2相关X蛋白(Bax)、抗癌基因p53(p53)、细胞色素C(Cyto-C)、凋亡诱导因子(AIF)蛋白表达。结果模型组细胞核染色质浓聚,线粒体肿胀,粗面内质网扩张,呈凋亡细胞改变;黑绛丹组和对照1组细胞线粒体结构正常,见胶原纤维,桥粒丰富;对照2组细胞核染色质浓缩,粗面内质网扩张。与模型组比较,黑绛丹组创面组织Fas、Bax、Caspase-3、Caspase-8、Caspase-9、Cyto-C及AIF表达显著降低,Bcl-2表达显著升高(P<0.01)。与对照1组比较,黑绛丹组Fas、Caspase-8 p53、Caspase-3、Bcl-2、Cyto-C、Bax表达显著升高(P<0.05或(P<0.01);与对照2组比较,黑绛丹组Caspase-8、Bax、Caspase-9、Cyto-C、AIF表达降低,Bcl-2、Fas L表达升高(P<0.05或(P<0.01)。结论黑绛丹可通过抑制局部创面组织细胞凋亡、调节凋亡相关蛋白表达从而减轻大鼠放射性皮炎损伤程度。
Objective To observe the possible mechanism of Hei-Dan treatment of radiation dermatitis. Methods Fifty-five rats were irradiated with 60Co γ-rays to establish a fourth-degree radiodermatitis animal model. Forty-eight rats were randomly divided into three groups: control group, control group, model group, Twelve groups were treated with Hei Dan solution, topical rhEGF solution, water-soluble vitamin E and normal saline 1 ml once daily for 10 days. Another set of blank group 12, to saline model group rats the same site. Transmission electron microscopy was used to observe the ultrastructure of wound tissue in each group. The apoptosis-related factors (Fas, Fas L, Cysteine Aspartate Caspase-3, Caspase-8, Caspase-9, Bcl-2, , B-cell lymphoma 2 (Bax), p53 (p53), Cyto-C and AIF. Results In the model group, chromatin condensation, mitochondria swelling and rough endoplasmic reticulum were dilated and apoptotic cells were changed. The mitochondrial structure of Heladan Dan group and control group 1 was normal, with collagen fibers and desmosomes abundant. In control group 2, Chromatin condensation, rough endoplasmic reticulum expansion. Compared with the model group, the expression of Fas, Bax, Caspase-3, Caspase-8, Caspase-9, Cyto-C and AIF were significantly decreased and the expression of Bcl-2 was significantly increased in the Hepatic Dan group (P <0.01). Compared with the control group, the expressions of Fas, Caspase-8, Caspase-3, Bcl-2, Cyto-C and Bax were significantly increased (P <0.05 or P <0.01) , The expression of Caspase-8, Bax, Caspase-9, Cyto-C, AIF in Hepitane group decreased and the expression of Bcl-2 and Fas L increased (P <0.05 or P <0.01) Local wound tissue apoptosis, regulating apoptosis-related protein expression and thus reduce the degree of radiation injury in rats.