| Radiation Induced Lung Injury(RILI)is the result of rays hitting the normal lung tissue during chest tumor's radiotherapy process.Currently the negative impacts of radiation-induced lung injury treatments are quite high.As a result,finding cures from the traditional Chinese medicine perspective is highly meaningful.Transforming growth factor-?(TGF-?)-is a type of peptide that can regulate cells' growth and differentiation.Research shows that after radiotherapy,the plasma TGF-?1 water increase can be considered as the predictor for radiation-induced lung injury.Through X-rays on Mice alveolar type ? cells(AEC ?),radiation injury can be induced to observe Chinese traditional medicine maxing shigan decoction compound's effects on radiated Mice AEC ? cells proliferation rate,apoptosis rate,TGF?-1?smad2?smad3?smad6?smad7 mRNA,phosphorylation smad2,smad3,non-phosphorylated smad6,and smad7 protein after 24 hours,48 hours,and 72 hours.Simultaneously,to investigate maxing shigan decoction's other resistance mechanisms for radiation-induced lung injury.Methods:1.Serum preparation:SD rats 18,200-250g,Ma Xing Shi Gan Tang made crude drug concentration 2.44g/ml,Give medicine twice a day for a total of three days.After one to two hours of the last medicine injection,extract blood.2.Cell radiation injury model construction:Use 3.64Gy/min,hit Mice AEC ? with 8Gy dose of X-rays,which induces radiation injury on Mice AEC ?.3.Specimen treatment and observation methods:Use 5%,10%,15%of blood serum compounds group and normal mice blood serum group and intervene with Mice SEC ? respectively.Examine 24 hours,48 hours,72 hours-every time period's AEC ? water level increase and post 48 hours apoptosis rate.RT-PCR examine AEC II's internal TGF-?1?Smad2?Smad3?Smad6 and Smad7 mRNA level.Western Blot examine AEC ?'s internal TGF-?/Smad signal transduction pathway downstream signaling molecules phosphorylated Smad2,Smad3,non-phosphorylated Smad6 and Smad7 protein level.Result:1.Mice AEC II's proliferation result shows:traditional medicine's low,medium,and high dose when intervened after 24 hours,48 hours,and 72 hours,Mice AEC ?'s energy level decreased compared to normal serum in rats(P<0.01).At the 24 hours point,energy levels were reduced as the blood serum increased in concentration.At the 48 hours point,blood serum with high dose reduced in energy level(P<0.01),no significant difference between medium and high doses.At the 72 hours point,blood serum with medium dose decreased and high dose energy levels decreased(P<0.01),there were no significant difference between high and low doses.2.Mice AEC ? apoptosis result shows:traditional medicine's low,medium,and high doses when intervened after 48 hours,each groups apoptosis rate was higher than normal serum group(P<0.01);no significant difference amongst the groups themselves.3.The Result of RT-PCR:The smad2 mRNA genetic expression of traditional medicine's low,medium,and high group is lower than normal serum group(P<0.05);The smad2 mRNA genetic expression of traditional medicine's medium group is lower than traditional medicine's low group(P<0.05),and no significant difference between traditional medicine's medium group and low group.NO significant difference between the smad3 mRNA genetic expression of traditional medicine's low,medium,and high group.The smad6 mRNA genetic expression of traditional medicine's medium,and high group is higher than normal serum group(P<0.05),The smad6 mRNA genetic expression of normal serum group is lower than the control group(P<0.05),The smad6 mRNA genetic expression of traditional medicine's medium,and high group is higher than traditional medicine's low group,but these is no significant difference between them;The smad7 mRNA genetic expression of traditional medicine's medium,and high group is higher than normal serum group(P<0.05),NO significant difference between traditional medicine's medium group and normal serum group,The smad7 mRNA genetic expression of traditional medicine's medium,and high is higher group than traditional medicine's low group,but these is no significant difference between them;The TGF-?1 mRNA genetic expression of traditional medicine's low,medium,and high group is lower than normal serum group(P<0.05),but these is no significant difference between them.4.The Result of Western-blot:The phosphorylated Smad2 protein level of traditional medicine's low,medium,and high group is lower than normal serum group(P<0.05),traditional medicine's low,medium,and high group with increasing concentration of phosphorylated Smad2 protein levels were reduced(P<0.05).The phosphorylated Smad3 protein level of traditional medicine's low,medium,and high group is lower than normal serum group(P<0.05),the phosphorylated Smad3 protein level of traditional medicine's medium,and high group is lower than traditional medicine's low group(P<0.05),but these is no significant difference between them;The non-phosphorylated Smad6 protein level of traditional medicine's medium,and high group is higher than normal serum group(P<0.05),NO significant difference between medicine's low group and normal serum group.Traditional medicine's low,medium,and high group with increasing concentration of phosphorylated Smad6 protein levels were raise(P<0.05).The non-phosphorylated Smad7 protein level of traditional medicine's medium,and high group is higher than normal serum group(P<0.05),The non-phosphorylated Smad7 protein level of traditional medicine's low group is lower than normal serum group(P<0.05),Traditional medicine's low,medium,and high group with increasing concentration of phosphorylated Smad7 protein levels were raise(P<0.05).Conclusion:traditional medicine can Significantly inhibit the Proliferation of radiation injuryed AEC ?,promote the apoptosis of it,and inhibit the level of TGF-?1 Smad2 mRNA,promote the level of smad6 and smad7 mRNA.down the expression protein of smad2 and smad3,Elevated the expression protein of smad6 and smad7. |
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