J Ethnopharmacol:丹灯通脑胶囊对脑微血管内皮细胞的保护作用

2022-09-21 August MedSci原创

丹灯通脑胶囊为中成药,在我国长期用于治疗脑缺血性卒中,保护脑微血管内皮细胞是主要治疗策略。但 DDTNC 保护 BMECs 的整体化学信息和潜在生物活性成分及其潜在机制仍不清楚。

脑卒中是一种常见的脑血管疾病,是导致人类死亡或瘫痪的第二大原因,脑缺血性脑卒中(IS)占脑卒中发病率的 85% 以上,在世界范围内发病率很高。在IS的发生过程中,血脑屏障(BBB)的破坏被认为是IS的主要发病机制。BMECs是血脑屏障的重要组成部分之一,在维护血脑屏障的结构和功能方面发挥着至关重要的作用。缺血再灌注引起的氧化应激会导致活性氧(ROS)过表达和细胞内钙超载。过量的 ROS 和细胞内钙会引起线粒体的损伤和 DNA 的裂解。Wnt/β-catenin 是脑血管的基本通路,与神经元和脑血管细胞的再生有关,激活 Wnt/β-catenin 通路有助于维持 BBB 的完整性。

中药 (TCM) 具有多成分、多靶点的优势。丹灯通脑胶囊(DDTNC)是一种中成药,因其活血化瘀的优点被广泛用于治疗IS。DDTNC由4种中药组成,包括丹参、灯盏花、川芎和葛根。既往研究表明,DDTNC 可预防 MCAO 大鼠脑缺血再灌注损伤,DDTNC-CSF 可提高 OGD/R 损伤的脑微血管内皮细胞(BMECs)的活力。但DDTNC的物质基础和药理作用机制尚不清楚。

研究目的鉴定DDTNC的活性成分,探讨DDTNC对Wnt/β-catenin通路相关BMECs的保护作用。

材料和方法采用高效液相色谱结合二极管阵列检测器(HPLC-DAD)和超高效液相色谱结合四极杆飞行时间质谱法检测滴滴涕成分和含有滴滴涕成分的脑脊液(DDTNC-CSF)。 UPLC-Q-TOF-MS),分别。建立大脑中动脉闭塞实验大鼠模型(MCAO),通过 Longa 法和 TTC 染色评估 DDTNC 的治疗效果。通过免疫荧光染色测定脑微血管密度。使用氧-葡萄糖剥夺再灌注(OGD/R)引起的损伤BMECs来评价含有DDTNC组合物的脑脊液(DDTNC-CSF)的保护作用。细胞存活率采用CCK-8法检测,细胞内Ca 2+和活性氧(ROS)采用Fluo-3法估算。此外,通过Western blotting检测了Bax、Bcl-2、Wnt、β-catenin、GSK-3β的蛋白。

结果所有方法学研究的 RSD 值均小于 3.0%。在优化的 HPLC-DAD 色谱条件下共检测到20 种化合物。在UPLC-Q-TOF-MS阴性模式下,DDTNC-CSF中检测到峰1和峰2,分别鉴定为丹参素和葛根素。在UPLC-Q-TOF-MS正模式下,DDTNC-CSF中检测到峰1和峰3,分别鉴定为丹参素和灯盏花乙素。DDTNC显着降低了MCAO大鼠的Longa值和梗死体积,显着增加了脑微血管密度。细胞内Ca2+DDTNC-CSF组OGD/R损伤的BMECs中ROS显着降低。DDTNC-CSF可上调Bcl-2、β-catenin、wnt-1的表达,DDTNC-CSF可下调Bax和GSK3β的表达。

 

综上所述,该研究建立了DDTNC的HPLC-DAD指纹图谱,然后收集DDTNC给药7天的大鼠脑脊液,并采用UPLC-Q-TOF-MS进行分析。DDTNC-CSF中检测到丹参素、葛根素和灯盏花乙素,这可能是DDTNC核心疗效的物质基础。此研究为揭示DDTNC治疗IS的作用机制提供了科学依据,DDTNC有望成为治疗IS的有效药物。

 

原文:Huang Q, Qi J, Gao Z, Li L, Wang N, Seto S, Yao M, Zhang Q, Wang L, Tong R, Chen Y, Chen X, Hou J. Chemical composition and protective effect of cerebrospinal fluid of Dan-Deng-Tong-Nao capsules on brain microvascular endothelial cells injured by OGD/R. J Ethnopharmacol. 2022 Jan 30;283:114705. doi: 10.1016/j.jep.2021.114705. Epub 2021 Oct 14. PMID: 34655669.

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    2022-10-26 yb6560
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  5. [GetPortalCommentsPageByObjectIdResponse(id=1908744, encodeId=02071908e44f2, content=<a href='/topic/show?id=a8d0141605e' target=_blank style='color:#2F92EE;'>#pharma#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=47, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=14160, encryptionId=a8d0141605e, topicName=pharma)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=f634250, createdName=chentianping, createdTime=Tue Feb 07 04:37:28 CST 2023, time=2023-02-07, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1788510, encodeId=83121e885106b, content=<a href='/topic/show?id=38e014162e2' target=_blank style='color:#2F92EE;'>#Pharmacol#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=44, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=14162, encryptionId=38e014162e2, topicName=Pharmacol)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=930e243, createdName=dingxiaobo, createdTime=Tue Sep 20 16:37:28 CST 2022, time=2022-09-20, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1996518, encodeId=5ed21996518be, content=<a href='/topic/show?id=ed881120ec8' target=_blank style='color:#2F92EE;'>#MAC#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=33, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=11207, encryptionId=ed881120ec8, topicName=MAC)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4b65161, createdName=yb6560, createdTime=Wed Oct 26 05:37:28 CST 2022, time=2022-10-26, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1936610, encodeId=dc85193661075, content=<a href='/topic/show?id=47be895549f' target=_blank style='color:#2F92EE;'>#血管内皮细胞#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=38, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=89554, encryptionId=47be895549f, topicName=血管内皮细胞)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=7a5c2500147, createdName=12498bb3m90(暂无昵称), createdTime=Fri Aug 18 17:37:28 CST 2023, time=2023-08-18, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1787838, encodeId=e6e21e878389d, content=<a href='/topic/show?id=fa03141591e' target=_blank style='color:#2F92EE;'>#Pharm#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=36, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=14159, encryptionId=fa03141591e, topicName=Pharm)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=8942242, createdName=jj000001, createdTime=Mon Oct 10 22:37:28 CST 2022, time=2022-10-10, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1365455, encodeId=1ce0136545539, content=<a href='/topic/show?id=2a2150e76fd' target=_blank style='color:#2F92EE;'>#微血管#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=50776, encryptionId=2a2150e76fd, topicName=微血管)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=669d219, createdName=mgqwxj, createdTime=Sat Sep 17 14:37:28 CST 2022, time=2022-09-17, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1599195, encodeId=e8d515991955f, content=<a href='/topic/show?id=f14d2e187ac' target=_blank style='color:#2F92EE;'>#保护作用#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=33, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=27187, encryptionId=f14d2e187ac, topicName=保护作用)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=2ee918684940, createdName=photoman, createdTime=Sat Sep 17 14:37:28 CST 2022, time=2022-09-17, status=1, ipAttribution=)]
    2022-10-10 jj000001
  6. [GetPortalCommentsPageByObjectIdResponse(id=1908744, encodeId=02071908e44f2, content=<a href='/topic/show?id=a8d0141605e' target=_blank style='color:#2F92EE;'>#pharma#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=47, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=14160, encryptionId=a8d0141605e, topicName=pharma)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=f634250, createdName=chentianping, createdTime=Tue Feb 07 04:37:28 CST 2023, time=2023-02-07, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1788510, encodeId=83121e885106b, content=<a href='/topic/show?id=38e014162e2' target=_blank style='color:#2F92EE;'>#Pharmacol#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=44, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=14162, encryptionId=38e014162e2, topicName=Pharmacol)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=930e243, createdName=dingxiaobo, createdTime=Tue Sep 20 16:37:28 CST 2022, time=2022-09-20, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1996518, encodeId=5ed21996518be, content=<a href='/topic/show?id=ed881120ec8' target=_blank style='color:#2F92EE;'>#MAC#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=33, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=11207, encryptionId=ed881120ec8, topicName=MAC)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4b65161, createdName=yb6560, createdTime=Wed Oct 26 05:37:28 CST 2022, time=2022-10-26, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1936610, encodeId=dc85193661075, content=<a href='/topic/show?id=47be895549f' target=_blank style='color:#2F92EE;'>#血管内皮细胞#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=38, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=89554, encryptionId=47be895549f, topicName=血管内皮细胞)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=7a5c2500147, createdName=12498bb3m90(暂无昵称), createdTime=Fri Aug 18 17:37:28 CST 2023, time=2023-08-18, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1787838, encodeId=e6e21e878389d, content=<a href='/topic/show?id=fa03141591e' target=_blank style='color:#2F92EE;'>#Pharm#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=36, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=14159, encryptionId=fa03141591e, topicName=Pharm)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=8942242, createdName=jj000001, createdTime=Mon Oct 10 22:37:28 CST 2022, time=2022-10-10, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1365455, encodeId=1ce0136545539, content=<a href='/topic/show?id=2a2150e76fd' target=_blank style='color:#2F92EE;'>#微血管#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=50776, encryptionId=2a2150e76fd, topicName=微血管)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=669d219, createdName=mgqwxj, createdTime=Sat Sep 17 14:37:28 CST 2022, time=2022-09-17, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1599195, encodeId=e8d515991955f, content=<a href='/topic/show?id=f14d2e187ac' target=_blank style='color:#2F92EE;'>#保护作用#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=33, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=27187, encryptionId=f14d2e187ac, topicName=保护作用)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=2ee918684940, createdName=photoman, createdTime=Sat Sep 17 14:37:28 CST 2022, time=2022-09-17, status=1, ipAttribution=)]
    2022-09-17 mgqwxj
  7. [GetPortalCommentsPageByObjectIdResponse(id=1908744, encodeId=02071908e44f2, content=<a href='/topic/show?id=a8d0141605e' target=_blank style='color:#2F92EE;'>#pharma#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=47, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=14160, encryptionId=a8d0141605e, topicName=pharma)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=f634250, createdName=chentianping, createdTime=Tue Feb 07 04:37:28 CST 2023, time=2023-02-07, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1788510, encodeId=83121e885106b, content=<a href='/topic/show?id=38e014162e2' target=_blank style='color:#2F92EE;'>#Pharmacol#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=44, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=14162, encryptionId=38e014162e2, topicName=Pharmacol)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=930e243, createdName=dingxiaobo, createdTime=Tue Sep 20 16:37:28 CST 2022, time=2022-09-20, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1996518, encodeId=5ed21996518be, content=<a href='/topic/show?id=ed881120ec8' target=_blank style='color:#2F92EE;'>#MAC#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=33, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=11207, encryptionId=ed881120ec8, topicName=MAC)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4b65161, createdName=yb6560, createdTime=Wed Oct 26 05:37:28 CST 2022, time=2022-10-26, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1936610, encodeId=dc85193661075, content=<a href='/topic/show?id=47be895549f' target=_blank style='color:#2F92EE;'>#血管内皮细胞#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=38, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=89554, encryptionId=47be895549f, topicName=血管内皮细胞)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=7a5c2500147, createdName=12498bb3m90(暂无昵称), createdTime=Fri Aug 18 17:37:28 CST 2023, time=2023-08-18, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1787838, encodeId=e6e21e878389d, content=<a href='/topic/show?id=fa03141591e' target=_blank style='color:#2F92EE;'>#Pharm#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=36, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=14159, encryptionId=fa03141591e, topicName=Pharm)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=8942242, createdName=jj000001, createdTime=Mon Oct 10 22:37:28 CST 2022, time=2022-10-10, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1365455, encodeId=1ce0136545539, content=<a href='/topic/show?id=2a2150e76fd' target=_blank style='color:#2F92EE;'>#微血管#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=50776, encryptionId=2a2150e76fd, topicName=微血管)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=669d219, createdName=mgqwxj, createdTime=Sat Sep 17 14:37:28 CST 2022, time=2022-09-17, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1599195, encodeId=e8d515991955f, content=<a href='/topic/show?id=f14d2e187ac' target=_blank style='color:#2F92EE;'>#保护作用#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=33, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=27187, encryptionId=f14d2e187ac, topicName=保护作用)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=2ee918684940, createdName=photoman, createdTime=Sat Sep 17 14:37:28 CST 2022, time=2022-09-17, status=1, ipAttribution=)]