基于PI3K/AKT/iNOS通路探究电针对干眼炎症反应的干预机制OA

Objective To investigate the therapeutic mechanism of electroacupuncture in dry eye disease in guinea pigs through the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/inducible nitric oxide synthase(iNOS)pathway axis.Methods Twenty-four British short-haired tricolor guinea pigs were randomly assigned to the blank group,model group,western medicine group,and electroacupuncture group,with 6 guinea pigs in each group.The blank group re-ceived no treatment,while dry eye models were induced in the other groups by subcutaneous injection of scopolamine hydrobromide for 35 consecutive days.The western medicine group was treated with fluorometholone eye drops bilaterally,and the electroacupuncture group received electroacupuncture at bilateral acupoints including Jingming,Taiyang,Cuanzhu,Tongziliao,and Sizhukong.Both the western medicine group and the electroacupuncture group were treated for 14 consecu-tive days.Tear film break-up time(BUT),corneal fluorescein staining(FL),and Schirmer I test(SIT)were measured at day 0,day 22,and day 35 after intervention.Hematoxylin-eosin staining was used to evaluate the histomorphology of the lac-rimal gland and conjunctival tissues under light microscopy.The degree of inflammatory cell infiltration in the conjunctival tissue was analyzed using the integral optical density method.Western blot was performed to detect the protein expression of PI3K,AKT,phosphorylated AKT(p-AKT),and iNOS in the lacrimal gland of guinea pigs.Quantitative real-time polymer-ase chain reaction was employed to measure the relative mRNA expression of iNOS,interleukin-6(IL-6),and tumor necro-sis factor-α(TNF-α).Results At day 35 after intervention,the model group exhibited shortened BUT,elevated FL score,and reduced SIT compared with the blank group,and the differences were statistically significant(all P＜0.05).The west-ern medicine group and the electroacupuncture group showed prolonged BUT,decreased FL score,and increased SIT rela-tive to the model group,and the differences were statistically significant(all P＜0.05).Under light microscopy,the model group displayed marked histopathological changes in the lacrimal gland and conjunctiva,compared with the blank group.Compared with the model group,the western medicine group and the electroacupuncture group demonstrated alleviated tis-sue injury in the lacrimal gland and conjunctiva.The results of integral optical density analysis showed that compared with the blank group,the inflammatory cell infiltration in the conjunctival tissue of guinea pigs in the model group was aggrava-ted,and the difference was statistically significant(P＜0.05).Compared with the model group,the inflammatory cell infil-tration in the conjunctival tissue of guinea pigs in the electroacupuncture group was alleviated,and the difference was statis-tically significant(P＜0.05).According to Western blot results,lacrimal gland tissues from the model group showed signifi-cantly upregulated protein expression of PI3K,p-AKT/AKT,and iNOS,alongside elevated mRNA levels of iNOS,IL-6,and TNF-α,compared with the blank group,and the differences were statistically significant(all P＜0.05).The western medi-cine group and the electroacupuncture group exhibited downregulated PI3K,p-AKT/AKT,and iNOS protein levels,as well as reduced mRNA expression of iNOS,IL-6,and TNF-α,relative to the model group,and the differences were statistically signif-icant(all P＜0.05).Conclusion Electroacupuncture mitigates lacrimal gland inflammation in guinea pigs with dry eye,suppresses the overactivation of the PI3K/AKT pathway,and lowers the transcriptional levels of downstream inflammatory markers,including iNOS,IL-6,and TNF-α.