局部外敷刺山柑总生物碱乳膏的系统性硬皮病小鼠皮肤胶原沉积情况观察

Observation of collagen deposition in the skin of mice with systemic scleroderma treated with capers total alkaloid cream

目的观察刺山柑总生物碱乳膏局部外敷系统性硬皮病(SSc)小鼠皮肤胶原沉积情况。方法48只小鼠随机分为刺山柑组、青霉胺组、模型组、正常组各12只,正常组皮下注射生理盐水,其余各组小鼠背部皮下注射盐酸博来霉素(30 mg/d)制备SSc模型,制模成功后,刺山柑组背部注射部位外敷500 mg/kg刺山柑总生物碱乳膏,青霉胺组灌胃给予125 mg/kg青霉胺,模型组及正常组外敷不含药物乳膏基质。采用Masson染色法观察各组小鼠胶原蛋白沉积情况,并计算胶原容积分数;采用蛋白印迹法检测小鼠皮肤组织Toll样受体4(TLR4)、髓样分化因子88(MyD88)、细胞核因子(NF-κB)蛋白,免疫组化检测小鼠皮肤组织TLR4、MyD88蛋白。结果与正常组比较,模型组小鼠胶原蛋白沉积显著增加(P<0.05);与模型组比较,刺山柑组及青霉胺组胶原蛋白沉积明显减少(P均<0.05)。与正常组比较,模型组小鼠TLR4、MyD88、NF-κB蛋白表达增高(P均<0.05);与模型组比较,刺山柑组及青霉胺组TLR4、MyD88、NF-κB蛋白表达降低(P均<0.05)。结论刺山柑总生物碱可以减少SSc小鼠皮肤胶原沉积,机制可能与抑制TLR4/MyD88通路相关。

Objective To observe the skin collagen deposition in mice with systemic scleroderma(SSc)treated with capers total alkaloid cream.Methods Forty-eight mice were randomly divided into the capers group,penicillamine group,model group,and normal group,with 12 mice in each.Mice in the normal group were injected with normal saline subcutaneously,and mice in the other groups were injected with bleomycin hydrochloride(30 mg/d)subcutaneously to make SSc models.After successful modeling,mice in the capers group were given 500 mg/kg capers total alkaloid cream at the injection site,mice in the penicillamine group were given 125 mg/kg penicillamine,and mice in the model group and normal group were given drug-free cream matrix.Masson staining was used to observe collagen deposition in each group,and collagen volume fraction was calculated.The proteins of toll-like receptor 4(TLR4),myeloid differentia⁃tion factor 88(MyD88),and nuclear factor NF-κB were detected by Western blotting,and TLR4 and MyD88 protein were detected by immunohistochemistry.Results Compared with the normal group,collagen deposition significantly in⁃creased in the model group(P<0.05).Compared with the model group,collagen deposition significantly decreased in the capers group and penicillamine group(both P<0.05).Compared with the normal group,TLR4,MyD88 and NF-κB pro⁃tein expression levels increased in the model group(all P<0.05).Compared with the model group,the expression levels of TLR4,MyD88,and NF-κB protein in the capers group and penicillamine group decreased(all P<0.05).Conclusion Caper alkaloids can reduce skin collagen deposition in SSc mice,and its mechanism may be related to the inhibition of TLR4/MyD88 pathway.