R285.5
杭州市卫生科技计划重点项目(2014Z08)
目的:从自噬相关因子的表达研究加味麻杏石甘汤干预放射性肺损伤的发生机制。方法:将肺泡细胞 A549 分为空白组、模型组及麻杏石甘汤低、中、高剂量组(以下简称低、中、高剂量组)。除空白组外,其余各组细胞进行 X 线照射,建立放射性肺损伤细胞模型。模型组细胞用 10%正常大鼠血清培养,低、中、高剂量组分别用 10%低、中、高剂量的含药血清培养。分别采用 Western-blot 及 RT-PCR 检测肺泡细胞 A549 中转化生长因子-β1 (TGF-β1)、Beclin-1、LC3 蛋白及 mRNA 表达。结果:与空白组比较,模型组细胞中 TGF-β1、Beclin-1、LC3 蛋白及 mRNA 表达均升高 (P<0.05)。与模型组比较,低、中、高剂量组细胞中 TGF-β1、Beclin-1、LC3 mRNA 表达均下降 (P<0.05),且随着剂量的升高而下降 (P<0.05);低、中、高剂量组 TGF-β1 蛋白表达下降(P<0.05),中、高剂量组 Beclin-1、LC3 蛋白表达下降(P<0.05),且 TGF-β1、Beclin-1、LC3 蛋白表达随着剂量的升高而下降 (P<0.05)。结论:加味麻杏石甘汤可通过下调肺泡细胞 A549 内 TGF-β1、LC3、Beclin-1 水平,调控自噬过程,减轻放射性肺损伤。
Abstract:Objective:TodiscusstheoccurrencemechanismofmodifiedMaxingShigantanginterveningradiation- inducedlunginjuryfromtheperspectiveofexpressionofautophagy-relatedfactors. Methods:A549 alveolarcellswere dividedintotheblankgroup,themodelgroup,andtheMaxingShigantanggroupsoflow-dose,medium-dose,andhigh- dose(low-dose,medium-dose,andhigh-dosegroupsforshort).Exceptfortheblankgroup,intheothergroups,X-ray irradiationwasconducted,andcellmodelswithradiation-inducedlunginjurywereestablished.Inthemodelgroup,cells wereculturedwith 10%serumofnormalrats.Inthelow-dose,medium-dose,andhigh-dosegroups,cellswerecultured with 10%medicatedserumoflow-dose, medium-dose, andhigh-dose.TheexpressionofproteinandmRNAof transforminggrowthfactor- β1(TGF- β1),Beclin-1,andLC3 inA549 alveolarcellswasdetectedbyWestern-blotand RT-PCR. Results:Comparedwiththoseintheblankgroup,theexpressionofproteinandmRNAofTGF-β1,Beclin-1, andLC3 inthemodelgroupwasincreased(P<0.05).Comparedwiththoseinthemodelgroup,theexpressionofmRNAof TGF-β1,Beclin-1,andLC3 inlow-dose,medium-dose,andhigh-dosegroupswasdecreased(P<0.05),anditwas decreasedasthedosewasincreased(P<0.05).Comparedwiththoseinthemodelgroup,theexpressionofproteinofTGF-β 1 inlow-dose,medium-dose,andhigh-dosegroupswasdecreased(P<0.05);theexpressionofproteinofBeclin-1 and LC3 wasdecreasedinmedium-doseandhigh-dosegroups(P<0.05);theexpressionofproteinofTGF-β1,Beclin-1,and LC3 inlow-dose,medium-doseandhigh-dosegroupswasdecreasedasthedosewasincreased(P<0.05). Conclusion: ModifiedMaxingShigantangcanalleviateradiation-inducedlunginjurybydown-regulatinglevelsofTGF- β1,Beclin-1, andLC3 inA549 alveolarcells,aswellasregulatingautophagy.
林贤雷,钟亚珍,陆金华,林泽晨,林胜友.基于自噬途径探讨加味麻杏石甘汤干预放射性肺损伤的分子机制[J].新中医,2021,53(8):15-18