Effect of Jianghu Decoction on immune inflammatory injury in connective tissue disease associated interstitial lung disease model mice
[Objective]To investigate the effect of Jianghu Decoction on pulmonary interstitial lesions in bleomycin(BLM)-induced connective tissue disease associated interstitial lung disease,CTD-ILD model mice.[Methods]The mice model of ILD was established in SPF male C57BL/6J mice by intratracheal dripping of BLM(2 mg/kg).The 30 mice with successful modeling were randomly divided into model group,positive drug group,high,medium and low dose groups of Jianghu Decoction,6 mice in each group.Six male C57BL/6J mice were set as a normal group.Mice in the normal group and the model group were given deionized water[10 mL/(kg·d)].Mice in the positive group was given prednisone[7.5 mg/(kg·d)].Mice in the Jianghu Decoction groups were given Jianghu Decoction 26.66,13.33 and 6.67 g/(kg·d),respectively.All mice in each group were given intragastric administration once a day,but the Jianghu Decoction high dose group was given twice a day.Lung tissue samples were collected after 4 weeks of continuous intervention.Hematoxylin-eosin(HE)staining method was used to observe the pathological morphology of the lung tissue in each group of mice.The protein expression levels of high mobility group box1(HMGB1),transforming growth factor beta1(TGF-β1),nuclear factor kappa-Bp65(NF-κB p65),Adenosine monophosphate AMP-activated protein kinaseα1(AMPKα1),pAMPKα1 in lung tissue were detected by Western Blot.[Results]Compared with the normal control group,the mice in the model group had obvious pathological changes(P＜0.01),mainly manifested as inflammatory cell infiltration and interstitial fibrosis in lung tissue.The inflammatory lesions of lung tissue in the positive drug group were improved compared with the model group(P＜0.05).The infiltration of inflammatory cells and interstitial fibrosis in the lung tissue of mice in each treatment group of Jianghu Decoction were improved compared with the model group(P＜0.05).Western Blot results showed that compared with the normal group;the expression levels of HMGB1 and NF-κB p65 protein in the lung tissue of the model group were significantly increased(P＜0.01);the expression level of TGF-β1 protein was significantly increased(P＜0.05),and the expression levels of AMPKα1 and pAMPKα1 protein were significantly decreased(P＜0.05,P＜0.01).Compared with the model group,the expression levels of HMGB1 and NF-κB p65 protein in the lung tissue of mice in the middle and low dose groups of Jianghu Decoction were significantly decreased(P＜0.05).The expression level of TGF-[31 protein in the lung tissue of mice in the high dose group of Jianghu Decoction was significantly decreased(P＜0.05).The expression level of AMPKα1 protein in the lung tissue of mice in the positive drug group was significantly up-regulated(P＜0.05),and the expression level of pAMPKα1 protein in the lung tissue of mice in the high dose group of Jianghu Decoction was significantly up-regulated(P＜0.05).Compared with the positive drug group,the expression level of TGF-β1 protein in the lung tissue of mice in the high-dose Jianghu Decoction group was significantly down-regulated(P＜0.05).The expression level of TGF-β1 protein in the high dose group of Jianghu Decoction was significantly different from that in the middle dose group of Jianghu Decoction(P＜0.05).[Conclusion]The middle and low dose groups of Jianghu Decoction have more advantages in improving the inflammatory pathological injury of lung tissue in mice and inhibiting the high expression of immune inflammatory factors.The high-dose group of Jianghu Decoction has a stronger effect on improving the interstitial fibrosis of lung tissue in mice,inhibiting the high expression of fibrogenic factors and up-regulating the immune inflammatory inhibitory factors.The molecular mechanism may be related to the activation of AMPK,thereby inhibiting HMGB1/NF-κB/TGF-β1 immune inflammatory signaling pathway.
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