Study on anti-hepatic fibrosis effect of Mongolian medicine Qiwei Qinggan powder based on NF-κB signaling pathway
AIM To investigate the anti-hepatic fibrosis effect of Qiwei Qinggan powder(QGS-7)through NF-κB signaling pathway in vivo and in vitro.METHODS In vivo experiment,sixty male Wistar rats were randomly divided into 5 groups:blank group,CCl4 model group,QGS-7 low,medium,high dose groups.The blank group was gavaged with 0.5％CMC-Na solution daily,and the model group and each dose groups were gavaged with 50％CCl4 peanut oil solution twice a week,while each dose groups were gavaged with corresponding dose of QGS-7(135,270,405 mg·kg-1·d-1)for 8 weeks.The histopathological changes of liver were observed by HE and Masson staining.q-PCR and Western blot were used to detect the mRNA and protein expression of fibrosis markers of alpha smooth muscle actin(α-SMA),collagen Ⅰ and NF-κB signaling pathway-related factors.In vitro experiment,rats were gavage with QGS-7 1 350 mg·kg-1·d-1,and blood was taken after 7 d to prepare drug-containing serum,which was classified into the blank group,the LPS model group,and the QGS-7 drug-containing serum low-,medium-and high-concentration groups using HSC-T6 cells.Detection of apoptosis by double staining with Annexin V-FITC and PI.The mRNA and protein expression of α-SMA,collagen Ⅰ and NF-κB signaling pathway-related factors were detected by q-PCR and Western blot.RESULTS In vivo experiment,HE and Masson results showed that fibroblasts were increased of the tissues in the CCl4 model group,with disorganized cell arrangement and significant collagen deposition compared with the blank group.Compared with the blank group,α-SMA,collagen Ⅰ and NF-KBp65 protein and mRNA expressions of the liver tissues were significantly increased in the CCl4 model group(P＜0.05),and TLR4,p-TAK1,p-IKKα/IKKα,p-IKKβ/IKKβ,p-NF-KBp65/NF-κBp65 protein and mRNA expression were significantly increased(P＜0.05).Compared with the model group,the protein and mRNA expression of the above factors were significantly decreased in each dose group of QGS-7(P＜0.05).In vitro experiments,the apoptosis rate was lower in both the blank group and the LPS model group.Compared with the model group,the apoptosis rate was significantly increased in each concentration of the QGS-7-containing serum group(P＜0.01),and the mRNA and protein expression of the α-SMA,collagen Ⅰ,and the NF-κB signalling pathway related to factors were consistent with the trend in vivo experimental expression.CONCLUSION QGS-7 has good anti-fibrotic effects,and the mechanism may through down-regulation of key targets of NF-κB signaling pathway to exert anti-fibrotic effects.
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