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The inflammation was marked and suppressed in rats treated with YKF, prednisone, or pirfenidone from 7 to 70 days after BLM was challenged, as well as the decrease in Ashcroft scores for fibrosis (Figures 2(a), 2(b), and 3(a)).
Analyses of HYP contents were conducted to evaluate the effect of YKF. HYP was elevated in BLM challenged rats from day 28 up to day 70.
Compared with BLM group, lymphocyte level of YKF group decreased significantly on days 7,14, 28, and 70 (P < 0.05 or P < 0.01); lymphocyte population decreased significantly in YKF group (days 7, 14, and 42, P < 0.05 or P < 0.01), prednisone group (days 7,14, and 28, P < 0.05 or P < 0.01), and pirfenidone treated group (day 14, P < 0.01).
Compared with BLM group, the neutrophils level significantly decreased in YKF (on days 7, 14, and 28, P < 0.01), prednisone (on days 14 and 28, P < 0.05 or P < 0.01), and pirfenidone group (on days 7 and 28, P < 0.01).
Monocytes increased on days 7 and 14 in bleomycin challenged rats (P < 0.01) compared with BLM group; they were significantly decreased in YKF, prednisone, and pirfenidone groups on days 7 and 14 (P < 0.05 or P < 0.01) (Figure 4(d)).
From day 7 to day 70, the expression of TNF-[alpha] protein and mRNA markedly increased in BLM challenged rats (P < 0.01) and significantly decreased in YKF, prednisone, and pirfenidone treated rats (Figures 5(a)~5(c)).
To further clarify the role of YKF on BLM-induced pulmonary injury, activated proteins related to NF-[kappa]B signal pathway were detected by Western blotting.
In clinical treatment of IPF patients, we found that YKF had beneficial effects on alleviating the clinical symptoms of IPF patients.
The administration of YKF has many beneficial effects, such as improving FVC, alleviating pulmonary injuries, and reducing fibrosis degree, and it still has long-term effect after withdrawal.
Our results have shown that TNF-[alpha] was decreased in YKF treated rats lungs, and IFN-[gamma] was increased.
In this study, we focused on the phosphorylations of cytoplasm IKK[beta] and I[kappa]B[alpha] and nuclear NF-[kappa]B  and found that the levels of p- IKK[beta], p-I[kappa]B[alpha], and p-NF-[kappa]B were upregulated in bleomycin rats at each time point and were downregulated by the administration of YKF. The results above suggest that anti-inflammatory response and alleviating fibrosis effects of YKF contribute to its protective effect against pulmonary fibrosis.
Taken together, YKF has beneficial protective and long-term effects on pulmonary fibrosis by anti-inflammatory response and alleviating fibrosis.
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