当归四逆汤调控Hedgehog信号通路抑制硬皮病皮肤纤维化的机制研究OA

Objective To investigate the mechanism of Danggui Sini Decoction(DSD)on skin fibrosis in bleomycin-induced scleroderma mouse model based on the Hedgehog signaling pathway.Methods Forty-eight C57BL/6J mice were randomly divided into a blank control group,a model group,a positive control group,and low-,medium-,and high-dose DSD groups,with 8 mice in each group.Except for the blank control group,mince in all groups received subcutaneous injections of bleomycin hydrochloride to establish the scleroderma mouse model.After successful model establishment,mice in the blank control and model groups were gavaged with an equivalent volume of saline at a dose of 0.2 mL/10 g.Mice in positive control group were gavaged with prednisone acetate at a dose of 0.45 mg/(kg∙d).Mice in low-,medium-,and high-dose DSD groups were gavaged with DSD at doses of 6,12,and 24 g/(kg∙d)respectively.The administration was performed once daily for 28 consecutive days.General condition of the mice was observed;the pathological changes of the skin tissues were observed using HE and Masson's trichrome staining;the levels of inflammatory cytokines IL-4 and IL-6 were detected using ELISA,the protein expression of α-SMA and COL1A1 were analyzed using Western blot;the mRNA expression of the Hedgehog signaling pathway(SHH,SMO,GLI1,GLI2,PTC,and GLI3)were assessed using RT-PCR.Results Compared with the blank control group,mice in the model group displayed significant sclerotic changes in the skin;skin lesions showed improvement with various degrees in the positive control group,as well as the low-,medium-,and high-dose DSD groups.The ELISA results showed that,compared with the blank control group,serum levels of IL-4 and IL-6 were significantly elevated in the model group(P<0.05);compared with the model group,the levels of both IL-4 and IL-6 were significantly reduced in the positive control group and all the DSD groups(P<0.05).Western blot showed that,compared with the blank control group,the protein expression levels of α-SMA and COL1A1 were significantly higher in the model group(P<0.05);compared with the model group,the protein expression levels of α-SMA and COL1A1 were significantly reduced in the positive control group,as well as the medium-,and high-dose DSD groups(P<0.05);while there is no statistically significant difference in low-dose DSD group(P>0.05).RT-PCR results showed that,compared with the blank control group,the mRNA expression levels of SHH,SMO,GLI1,and GLI2 in the model group were significantly elevated(P<0.05),while the mRNA expression levels of PTC and GLI3 in the model group were reduced(P<0.05);Compared with the model group,the mRNA expression levels of SHH,SMO,GLI1,and GLI2 in the positive control group and all the DSD groups were significantly reduced(P<0.05);the mRNA expression levels of PTC and GLI3 were significantly increased in the positive control group,as well as the medium-,and high-dose DSD groups(P<0.05);while the low-dose DSD group showed no statistically significant difference in the mRNA expression of PTC and GLI3(P>0.05).Conclusion DSD can effectively alleviate skin fibrosis in scleroderma of mice and inflammation.Its mechanism may involve the inhibition of the Hedgehog signaling pathway,the downregulation of the expressions of α-SMA and COL1A1,and the suppression of inflammatory cytokine release,such as IL-4 and IL-6,ultimately exerting its anti-fibrotic action on the skin.