托里消毒散加减通过PI3K/Akt/mTOR通路调节炎症反应改善肉芽肿性乳腺炎的作用机制研究OA

Objective:To investigate the mechanism by which modified Tuoli Xiaodu San ameliorates granuloma-tous mastitis(GM)through regulation of inflammatory responses via the PI3K/Akt/mTOR signaling pathway.Methods:A rat GM model was established in SD rats by subcutaneous injection of a mixture of homogenized human GM lesion tissue and Freund's complete adjuvant.An additional 12 healthy rats were assigned to a normal control group(n=6)and an adjuvant control group(n=6,injected with Freund's adjuvant alone).Sixty rats with successful-ly induced GM were randomized into five groups(initial n=12 each):a model group,a methotrexate positive control group(0.35 mg/kg,once weekly),and low-,medium-,and high-dose Tuoli Xiaodu San groups(4.5,8.9,and 17.8 g/kg,respectively).Two rats died from procedural stress and were excluded according to pre-specified criteria,yielding a fi-nal sample size of n=10 per group.After 8 weeks of treatment,the mammary inflammation index was scored;his-topathological changes were examined by HE staining;Bax and Bcl-2 expression was assessed by immunohistochem-istry(×400);PI3K,p-PI3K,Akt,p-Akt,mTOR,and p-mTOR protein levels were measured by Western blot;corre-sponding mRNA levels were quantified by RT-qPCR;and serum IL-6,TNF-α,and IL-1β were detected by ELISA.Results:No significant differences were observed between the adjuvant control and normal groups for any measured index(P＞0.05),indicating that Freund's adjuvant alone did not elicit non-specific immune interference.Compared with the model group,all Tuoli Xiaodu San groups showed significantly reduced inflammation index scores(P＜0.05),with the greatest reduction in the high-dose group(P＜0.01).HE staining revealed attenuated inflammatory cell infiltration.Immunohistochemistry showed that,compared with the model group,the high-dose group had signifi-cantly lower Bax expression(P＜0.01)and higher Bcl-2 expression(P＜0.01),with the Bax/Bcl-2 ratio decreasing from 2.90 to 0.52.Western blot analysis revealed significant reductions in the p-PI3K/PI3K,p-Akt/Akt,and p-mTOR/mTOR ratios(P＜0.01),and ELISA showed marked decreases in serum IL-6,TNF-α,and IL-1β(P＜0.01).Conclusion:Modified Tuoli Xiaodu San may ameliorate GM by inhibiting activation of the PI3K/Akt/mTOR signaling pathway,down-regulating pro-inflammatory cytokines(IL-6,TNF-α,IL-1β),and modulating the Bax/Bcl-2 apoptotic balance,thereby improving the inflammatory microenvironment of mammary tissue.