逍遥散治疗大鼠糖尿病性勃起功能障碍的分析及实验  

Treatment of Diabetic Rats Mellitus-induced Erectile Dysfunction by Xiaoyaosan Based on Experiments

作　　者：卯银辉 王卓 孙军涛 魏志涛 王明星[2] 杨勇 MAO Yinhui;WANG Zhuo;SUN Juntao;WEI Zhitao;WANG Mingxing;YANG Yong(Changchun University of Chinese Medicine,Changchun 130117,China;The Affiliated Hospital of Changchun University of Chinese Medicine,Changchun 130021,China)

机构地区：[1]长春中医药大学,长春130117 [2]长春中医药大学附属医院,长春130021

出　　处：《中国实验方剂学杂志》2024年第17期122-130,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：吉林省自然科学基金面上项目(YDZJ202301ZYTS138);吉林省卫生健康科技能力提升项目(2022JC048)。

摘　　要：目的:探讨逍遥散治疗大鼠糖尿病性勃起功能障碍(DMED)的作用机制及其主要的活性成分。方法:采用链脲佐菌素(STZ)诱导糖尿病大鼠模型,通过海绵体内压/平均动脉压(ICP/MAP)的测量及马松染色对逍遥散进行疗效评价。然后,通过网络药理学和分子对接预测逍遥散治疗DMED的主要活性成分、关键靶点和潜在的信号通路。最后,对预测的结果进行体内外实验验证。结果:ICP/MAP和马松染色的检测结果表明,与正常组比较,模型组大鼠的勃起功能显著降低(P<0.01),平滑肌/胶原纤维显著降低(P<0.01);经过逍遥散治疗后,与模型组比较,糖尿病大鼠的ICP/MAP显著升高(P<0.01),平滑肌/胶原纤维显著升高(P<0.01)。网络药理学结果表明,逍遥散通过槲皮素、山柰酚、β-谷甾醇和豆甾醇等主要活性成分作用于白蛋白(ALB)、蛋白激酶B1(Akt1)、白细胞介素-6(IL-6)和肿瘤坏死因子(TNF)等核心靶点,参与糖尿病并发症中的晚期糖基化终末产物(AGE)/晚期糖基化终末产物受体(RAGE)信号通路和膦脂酰肌醇3-激酶(PI3K)/Akt信号通路的调节。分子对接结果显示,逍遥散的关键成分与核心靶点具有较好的结合能力,其中β-谷甾醇与ALB的结合能力最强。体内实验表明,逍遥散能够显著提高ALB和Akt1在血清中及mRNA的表达,抑制IL-6和TNF-α的表达;显著提高磷酸化(p)-PI3K和p-Akt的蛋白及mRNA的表达,抑制RAGE的表达。细胞热迁移实验(CETSA)结果表明,β-谷甾醇可以显著抑制ALB蛋白的降解。结论:逍遥散可能通过调节ALB、Akt1、IL-6、TNF等靶点及RAGE/PI3K/Akt信号通路恢复糖尿病大鼠的勃起功能,其主要的活性成分可能是β-谷甾醇。Objective:To investigate the mechanism of action and main active components of Xiaoyaosan in the treatment of diabetic mellitus-induced erectile dysfunction(DMED).Method:Streptozotocin(STZ)was used to induce a diabetic rat model.The therapeutic efficacy of Xiaoyaosan was evaluated by measuring intracavernous pressure/mean arterial pressure(ICP/MAP)and using Masson's trichrome staining.The main active components,key targets,and potential signaling pathways of Xiaoyaosan for the treatment of DMED were predicted by network pharmacology and molecular docking.The predicted results were then validated by in vitro and in vivo experiments.Result:The ICP/MAP measurements and Masson's staining results showed that compared with the results in the control group,the erectile function of rats in the model group was significantly reduced(P<0.01),and the ratio of smooth muscle/collagen fibers was significantly reduced(P<0.01).After treatment with Xiaoyaosan,compared with the results in the model group,the ICP/MAP value of the diabetic rats was significantly elevated(P<0.01),and the ratio of smooth muscle/collagen fibers was significantly higher(P<0.01).The results of network pharmacology showed that Xiaoyaosan acted on key targets such as albumin(ALB),protein kinase B1(Akt1),interleukin-6(IL-6),and tumor necrosis factor(TNF)through its main active components,including quercetin,kaempferol,β-sitosterol,and stigmasterol.These components were involved in the regulation of the advanced glycation end-products/receptor for advanced glycation end-products(AGE/RAGE)signaling pathway and the phosphoinositide 3-kinases(PI3K)/Akt signaling pathway in diabetic complications.The results of molecular docking showed that the key components of Xiaoyaosan had good binding capabilities with core targets,withβ-sitosterol showing the strongest binding affinity with ALB.In vivo experiments demonstrated that Xiaoyaosan could significantly increase the protein and mRNA expression of ALB and Akt1 in serum,and inhibit the expression of IL-6 and TNF-α

关 键 词：逍遥散 糖尿病性勃起功能障碍 网络药理学 分子对接 细胞热迁移 

分 类 号：R284[医药卫生—中药学] R285[医药卫生—中医学] R289R287R22R2-031R33R24