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纸质出版日期:2010
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殷宗宝, 王洪武, 邓超, 等. 甘利欣对肺纤维化大鼠的TGF-β、IFN-γ、IL-4的影响[J]. 中国实验方剂学杂志, 2010,16(6).
YIN Zong-bao, WANG Hong-wu, DENG Chao, et al. Effect of Glycyrrhizin on TGF-β、IFN-γ、IL-4 in Rat Model of Pulmonary Fibrosis[J]. Chinese journal of experimental traditional medical formulae, 2010, 16(6).
目的: 建立大鼠肺纤维化动物模型
探讨甘利欣治疗博莱霉素致肺纤维化大鼠的作用机制。 方法: 将SD级雄性大鼠60只
体重(180±20)g
随机分成4组:博莱霉素所致大鼠肺纤维化模型组(BLM组)、生理盐水对照组、甘利欣治疗组、地塞米松(Dexamethasone
DXM)治疗组
每组10只。博莱霉素致肺纤维化动物模型第28 d开始
各治疗组在每天同一时间段内连续14 d腹腔内注射。治疗的剂量分别为地塞米松注射液3 mg/(kg ·d)(浓度为5 mg/mL)、甘利欣150 mg(kg ·d);在治疗后第15 d取大鼠左肺下叶1 cm×1 cm×l cm大小的组织
固定于10%甲醛溶液
供免疫组化TGF-β1、IFN-γ、IL-4表达观察;同样取大鼠右肺下叶供HE染色病理观察。抽取大鼠6~8 mL血液静置30 min后中速离心15 min
取其血清用ELISA双抗体夹心法测定各组大鼠血清TGF-β1 IL-4、IFN-γ含量。 结果: ①采用分级方法定性评价HE染色大鼠肺组织的肺纤维化程度
发现甘利欣治疗组、DXM治疗组大鼠肺纤维化程度均较轻微。②模型组大鼠肺组织TGF-β1染色呈强阳性反应
IFN-γ染色较淡
而甘利欣治疗组、DXM治疗组TGF-β1、IFN-γ染色则相反。免疫组化TGF-β1定量分析结果显示:与模型组均数相比
对照组、地塞米松组、甘利欣组的P值分别为0.007
0.000
0.001;而甘利欣组与地塞米松组相比
P值为0.018。免疫组化IFN-γ定量分析:模型组与甘利欣组、地塞米松组相比
P值分别为0.004
0.010;而甘利欣组与地塞米松组相比
P值为0.618。③模型组大鼠血清TGF-β1
IL-4含量较高
IFN-γ含量较低
而甘利欣治疗组、DXM治疗组则相反。模型组大鼠血清TGF-β1均数与对照组、地塞米松组、甘利欣组相比
P值分别为0.008
0.044
0.005;地塞米松组与甘利欣组相比P值0.169。模型组大鼠血清IL-4均数与对照组、地塞米松组、甘利欣组相比
P值分别为0.013
0.029
0.013
地塞米松组、甘利欣组两两相比无差异(P>0.05)。地塞米松组、甘利欣组大鼠血清IFN-γ均数两两相比无差异(P=0.389)
但两者与模型组相比P值分别为0.001
0.000。 结论: 甘利欣可以减轻大鼠肺纤维化损伤和肺组织的TGF-β1、IL-4的表达
并降低他们在血清中的含量
增加血清中的IFN-γ含量
增强肺组织IFN-γ的表达
为临床治疗肺纤维化提供理论支持。
Objective: To investigate the mechanisms and effects of glycyrrhizin in rat model of pulmonary fibrosis produced by Bleomycin. Method: Sprague Dawley rats(180±20) g were randomLy divided into 4 groups: pulmonary fibrosis group
NS group
DXM treatment group
glycyrrhizin treatment group.There were ten rats in every group.All rats in the treatment groups were intraperitoneal injection after 28 days when rats were produced pulmonary fibrosis
which was at the same time every day and lasts for 14 days.The treatment dose were DXM 3 mg/(kg ·d)
glycyrrhizin dose 150 mg/(kg ·d).After the 15th days
Lung tissues were obtained
the left lung(1 cm×1 cm×lcm) was fixed in 10% formaldehyde
which were measured by immunity class TGF-β1
IFN-γ;the right lung was made into slice which cell apoptosis was detected.To draw 6~8 mL blood of the rats in all treatment groups and stand for 30 minutes
we can obtain blood-serum after the blood centrifuged for 15 mnutes
and the blood-serum war measured by ELISA TGF-β1、IFN-γ and IL-4. Result: 1.The extent of alveolitis and fibrosis of pulmonary fibrosis rats were quantified by grades. The pulmonary alveolitis and fibrosis of glycyrrhizin group and DXM group were signifieantly lighter than BLM group.2.Stronger TGF-β1 protein expression was deteeted in bronehiolar epitheliums of BLM group
poorer IFN-γ in BLM group.But the oppositions were in glycyrrhizin group and DXM group. TGF-β1 protein expression in BLM group was more significant deviation than in NS group(P=0.007)
DXM group(P=0.000)
glycyrrhizin group(P=0.001)
but the difference was obvious between DXM group and glycyrrhizin group(P=0.018); IFN-γ protein expressions in glycyrrhizin group(P=0.004) and in DXM group(P=0.01) were more significant deviation than in BLM group.But IFN-γ protein expression no deviation between DXM group and glycyrrhizin group(P=0.618);3.The TGF-β1and IL-4 levels of BLM group were higher than all the treatment groups
but the IFN-γ level was contrary. The TGF-β1 level of BLM group was more deviative than glycyrrhizin group(P=0.005)
DXM group(P=0.044)
NS group(P=0.008); The IL-4 levels of NS group(P=0.013)
DXM group(P=0.029)
glycyrrhizin group(P=0.013)were more deviative than in BLM group; The IFN-γ leve of BLM group was more poorer than DXM group(P=0.001)and glycyrrhizin group(P=0.000)
But the IFN-γ leve no deviation between DXM group and glycyrrhizin group(P=0.389). Conclusion: The study shows rats pulmonary fibrosis in the glycyrrhizin treatment group was less severe than that of BLM group. TGF-β1 and IL-4 protein expressions in the glycyrrhizin treatment group were poorer than in BLM group. The TGF-β1and IL-4 levels were the same.But IFN-γ protein expression and the IFN-γ leve in glycyrrhizin group were stronger.
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