Study on the mechanism of regulating Th17/Treg balance by Fuzheng Touxie Jiedu Decoction to improve lung injury in rats with Pseudomonas aeruginosa pneumonia
Objective To investigate the effect of Fuzheng Touxie Jiedu Decoction on lung injury in Pseudomonas aeruginosa pneumonia(PAP)rats and explore its possible mechanism.Methods The suspension of Pseudomonas aeruginosa(3×109 CFU/mL)0.1 mL was injected by orotracheal cannula to replicate the PAP rat model.A total of 36 SD rats were divided into the normal group,the model group,the Fuzheng Touxie Jiedu Decoction low-(3.18 g/kg),medium-(6.36 g/kg),high-(12.72 g/kg)dose groups and the piperacillin group(0.27g/kg)according to random number table method,with six rats in each group.Rats in each drug administration group received the corresponding drugs,while rats in the normal and model groups received an equal volume of distilled water by gavage once a day for seven consecutive days.Nucleic acid values of Pseudomonas aeruginosa were detected in the lung tissues of rats in each group by real-time fluorescence PCR.Hematoxylin-eosin staining was used to observe pathological morphological changes in lung tissues in rats in each group.Flow cytometry assay was used to detect the differentiation of T helper 17(Th17)cells and regulatory T(Treg)cells in the peripheral blood of rats in each group.Serum tumor necrosis factor-α(TNF-α),transforming growth factor-β(TGF-β),interleukin-6(IL-6),interleukin-10(IL-10),and interleukin-17(IL-17)levels were detected in rats of each group with an enzyme-linked immunosorbent assay.The mRNA level of retinoid-related orphan receptor γt(RORγt),forkhead box protein 3(FoxP3),and related inflammatory factors were detected by real-time fluorescence PCR.The protein expressions of RORγt,FoxP3,and related inflammatory factors were detected by Western blotting.Results The positive detection of Pseudomonas aeruginosa in the serum of model rats and the obvious inflammatory lesions observed in the pathological tissue sections indicated successful modeling.A large number of inflammatory cell infiltration,structural damage and epithelial cell shedding were observed in the lung issue of the model group rats.Compared to the normal group,the proportion of Th17 in CD4 T+cells of peripheral blood of rats in the model group increased,the proportion of Treg decreased and the ratio of Th17/Treg increased(P<0.01).The contents of TNF-α,TGF-β,IL-6,and IL-17 in serum were increased,while IL-10 was decreased(P<0.01).The mRNA expression levels of RORγt,TNF-α,TGF-β,IL-6,and IL-17 increased,while the mRNA expression levels of FoxP3 and IL-10 decreased in the model group(P<0.01).The protein expression levels of RORγt,TNF-α,TGF-β,IL-6,and IL-17 were increased,while those of FoxP3 and IL-10 were decreased(P<0.05,P<0.01).Compared to the model group,damage to lung structure,epithelial exfoliation condition,and inflammatory cell infiltration improved in the Fuzheng Touxie Jiedu Decoction low-,medium-,and high-dose groups and the piperacillin group.The proportion of Th17 decreased,the proportion of Treg increased,and the ratio of Th17/Treg decreased(P<0.05,P<0.01)in the Fuzheng Touxie Jiedu Decoction medium-and high-dose groups and the piperacillin group.Serum levels of TNF-α,TGF-β,IL-6,and IL-17 decreased,while anti-inflammatory factor(IL-10)levels increased in rats in all drug-administered groups(P<0.05,P<0.01).The mRNA expression levels of RORγt,TNF-α,TGF-β,IL-6,and IL-17 were decreased,the mRNA expression levels of FoxP3 and IL-10 were increased in the Fuzheng Touxie Jiedu Decoction medium-,high-dose groups and the piperacillin group(P<0.05,P<0.01).The protein expression levels of RORγt,TNF-α,TGF-β,IL-6,and IL-17 were decreased,the protein expression levels of FoxP3 and IL-10 were increased in the Fuzheng Touxie Jiedu Decoction medium-,high-dose groups and the piperacillin groups(P<0.01).Conclusion Fuzheng Touxie Jiedu Decoction showed a remarkable capability to alleviate lung inflammatory injury in PAP rat models,reduce the levels of proinflammatory factors,and enhance anti-inflammatory factors.Therapeutic efficacy is believed to operate through regulation of Th17/Treg immune balance.
Fuzheng Touxie Jiedu DecoctionPseudomonas aeruginosabacterial pneumoniahelper T cell 17/regulatory T cellinflammatory lung injuryrats
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