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目的用小鼠放射性肺损伤的模型来评价中药当归对放射性肺纤维化的防治作用。方法成年雌性 C57BL/6小鼠180只,按体重分为4组。空白对照组36只,腹腔注射生理盐水20ml·kg~(-1)·d~(-1);纯当归组36只,腹腔注射25%当归注射液20 ml·kg~(-1)·d~(-1);单纯照射组54只,全肺单次照射12 Gy+腹腔注射生理盐水20 ml·kg~(-1)·d~(-1);当归+照射组54只,全肺单次照射12 Gy+腹腔注射25%当归注射液20 ml·kg~(-1)·d~(-1)。于照射后1、24、72 h 和1、2、4、8、16、24周取各组小鼠肺组织,肺组织分别用 HE 和 Masson 染色在光镜下观察组织病理学改变和胶原纤维沉积,用碱水解法来测定羟脯氨酸含量和 RT-PCR 法来检测转化生长因子β1(transforming growth factor β1,TGF-β1)mRNA 相对含量。结果当归+照射组的肺组织与正常组织相似,未见明显纤维化病灶,其充血水肿较单纯照射组也明显减轻。单纯照射组小鼠的湿肺羟脯氨酸含量为(0.875±0.009)μg/mg,高于空白对照组(0.775±0.024)μg/mg 和单纯当归组(0.751±0.034)μg/mg,当归+照射组(0.782±0.010)μg/mg 低于单纯照射组(P<0.01)。RT-PCR 结果显示,单纯照射组的 TGF-β1 mRNA 相对含量(249.655±16.320)较单纯当归组(1.254±0.061)和空白对照组(1.324±0.057)差异均有统计学意义(P<0.01),而当归+照射组的 TGF-β1 mRNA 相对含量(108.076±9.870)低于单纯照射组(P<0.01)。结论成功的复制了小鼠放射性肺损伤的模型,并发现当归能明显降低放射性肺损伤中TGF-β1的表达水平,抑制胶原形成和减轻辐射所引起的病理变化,对小鼠的放射性肺纤维化可能具有一定的防治作用。
Objective To evaluate the preventive and therapeutic effects of Angelica sinensis on radiation-induced pulmonary fibrosis in a mouse model of radiation-induced lung injury. Methods 180 adult female C57BL/6 mice were divided into 4 groups according to body weight. Thirty-six rabbits in the control group received intraperitoneal injection of normal saline 20 ml·kg -1 ·d -1 ; 36 patients in the pure Angelica group received intraperitoneal injection of 25% Angelica Injection 20 ml·kg -1 ·d ~(-1); 54 animals in the simple irradiation group, 12% of whole lungs irradiated with 12 Gy+ saline 20 ml·kg~(-1)·d~(-1) intraperitoneally; 54 rats in the Angelica + irradiation group, whole lung alone The rats were irradiated with 12 Gy+ intraperitoneal injection of 25% Angelica injection 20 ml·kg~(-1)·d~(-1). Lung tissues from each group were taken at 1, 24, 72 h and 1, 2, 4, 8, 16 and 24 weeks after irradiation. Lung tissue was stained with HE and Masson under light microscope to observe histopathological changes and collagen fibers. Deposition, determination of hydroxyproline content by alkaline hydrolysis and RT-PCR were performed to determine the relative amount of transforming growth factor β1 (TGF-β1) mRNA. RESULTS: The lung tissues of the Angelica + irradiation group were similar to normal tissues, and no obvious fibrotic lesions were found. The hyperemia and edema of the lung tissue of the Angelica + irradiation group were also significantly reduced compared with the simple irradiation group. The content of hydroxyproline in the wet lung of mice in the simple irradiation group was (0.875±0.009) μg/mg, higher than that in the control group (0.775±0.024) μg/mg and that in the pure angelica group (0.751±0.034) μg/mg. The + irradiation group (0.782±0.010) μg/mg was lower than that of the simple irradiation group (P<0.01). RT-PCR results showed that the relative content of TGF-β1 mRNA (249.655±16.320) in the simple irradiation group was significantly higher than that in the pure Chinese angelica group (1.254±0.061) and the blank control group (1.324±0.057) (P<0.01). The relative content of TGF-β1 mRNA in the Angelica + irradiation group was lower (108.076±9.870) than that in the simple irradiation group (P<0.01). Conclusion The model of radiation-induced lung injury in mice was successfully replicated. It was found that Angelica sinensis can significantly reduce the expression of TGF-β1 in radiation-induced lung injury, inhibit the formation of collagen and reduce the pathological changes caused by radiation, and treat radiation-induced lung fibrosis in mice. May have a certain preventive effect.