腺苷A_(2A)受体拮抗剂通过PI3K途径抑制小鼠瘢痕增生的实验研究

Experimental Study on Adenosine A_(2A) Receptor Antagonist Inhibiting Scar Hyperplasia in Mice Through PI3K Pathway

目的:探究腺苷A_(2A)受体(A_(2A)R)拮抗剂SCH58261对小鼠瘢痕增生的影响,并进一步分析该影响作用与PI3K途径的相关性。方法:实验1:将32只BALB/c雄性小鼠随机分为4组(n=8)。各组均制备瘢痕增生模型,第5~14天时,对照组和模型组均用生理盐水局部注射、高低剂量组分别给2 mg/kg、5 mg/kgA;R拮抗剂SCH58261局部注射,每日1次、连续10 d。实验2:将32只BALB/c雄性小鼠随机分为4组(n=8)。各组制备瘢痕增生模型,第5~14天,Ad-NC组局部注射Ad-NC及生理盐水,AdNC+模型组局部注射Ad-NC及生理盐水、Ad-NC+高剂量组局部注射Ad-NC及5 mg/kgSCH58261局部注射、Ad-PI3K+高剂量组局部注射Ad-PI3K及5 mg/kgSCH58261局部注射,每日1次,连续10 d。采用HE染色观察增生瘢痕的组织学特征,采用Western blot检测转化生长因子-β_(1)(TGF-β_(1))、Ⅰ型胶原(Col-Ⅰ)、Ⅲ型胶原(Col-Ⅲ)、p-PI3K、p-AKT的表达水平。结果:实验1:模型组出现了典型的瘢痕增生,TGF-β_(1)、Col-I、Col-III、p-PI3K、p-AKT的表达水平均高于对照组;低剂量组和高剂量组瘢痕增生明显改善,TGF-β_(1)、Col-I、Col-III、p-PI3K、p-AKT的表达水平均低于模型组,差异有统计学意义(P<0.05);实验2:过表达PI3K后,高剂量SCH58261改善瘢痕增生及抑制TGF-β_(1)、Col-Ⅰ、Col-Ⅲ表达的作用发生逆转(P<0.05)。结论:腺苷A;R受体拮抗剂SCH58261能够抑制小鼠瘢痕增生,且这一作用与抑制PI3K途径有关。

Objective To investigate the effect of adenosine A_(2A)receptor(A_(2A)R) antagonist SCH58261 on scar hyperplasia in mice, and furtherly analyze the correlation between this effect and PI3K pathway. Methods Experiment 1: 32 BALB/C male mice were randomly divided into four groups(n=8). The scar hyperplasia model was prepared in each group. At 5~14 d, the control group and the model group were injected with normal saline, and the high and low dose groups were injected with 2mg/kg and 5mg/kg A;R antagonist SCH58261 respectively, once a day for 10 days. Experiment 2: 32 BALB/C male mice were randomly divided into four groups(n=8). The scar hyperplasia model was prepared in each group. At 5~14d, Ad-NC group was injected with Ad-NC and normal saline, ad Ad-NC+model group was injected with Ad-NC and normal saline, ad Ad-NC+high dose group was injected with Ad-NC and 5 mg/kg SCH58261, Ad-PI3K + high dose group was injected with Ad-PI3K and 5mg/kg SCH58261, once a day for 10 days.Then HE staining was used to observe the histological characteristics of hypertrophic scars,and western blot was used to detect transforming growth factor-β_(1)(TGF-β_(1)), type I collagen(Col-I), type III collagen(Col-III),p-PI3K, p-The expression level of AKT. Results Experiment 1: The model group had typical scar hyperplasia, the expression levels of TGF-β_(1), Col-I, Col-III, p-PI3K, and p-AKT were higher than those of the control group. scar hyperplasia was obvious in the low-dose and high-dose groups improved, the expression levels of TGF-β_(1), Col-I, Col-III, p-PI3K, and p-AKT were lower than the model group, the difference was statistically significant(P<0.05). Experiment 2: After overexpression of PI3K, high The effect of SCH58261 on improving scar hyperplasia and inhibiting the expression of TGF-β_(1), Col-I and Col-III was reversed(P<0.05). Conclusion The adenosine A;R receptor antagonist SCH58261 can inhibit scar hyperplasia in mice, and this effect is related to the inhibition of PI3K pathway.