基于“心受气于脾”探讨心痛泰调控p38 MAPK/AP-1对动脉粥样硬化兔的中医证候和VSMC胶原纤维的影响  

Effect of Xintongtai Regulating p38 MAPK/AP-1 on Traditional Chinese Medicine Syndrome Score and Collagen Fibers in VSMCs of Rabbits with Atherosclerosis:An Exploration Based on Theory of Heart Receiving Qi From Spleen

作　　者：易琼[1,2] 李雅 郭志华[2] 唐云[1] 申思[1] 齐婧[1] 魏佳明[2] YI Qiong;LI Ya;GUO Zhi-hua;TANG Yun;SHEN Si;QI Jing;WEI Jia-ming(The First Affiliate Hospital,Hunan University of Chinese Medicine,Changsha 410007,China;Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学第一附属医院,长沙410007 [2]湖南中医药大学,长沙410208

出　　处：《中国实验方剂学杂志》2021年第19期56-65,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：湖南省中医药科研项目重点课题(2021029);湖南省教育厅科学研究重点项目(20A384);湖南省自然科学基金面上项目(2021JJ30492);湖南省自然科学基金青年项目(2021JJ40426);湖南中医药大学中医学一流学科开放基金一般项目(2021ZYX19);湖南中医药大学中医学一流学科开放基金重点项目(2021ZYX14);湖南中医药大学中医学国内一流建设学科项目(湘教通[2018]469号)。

摘　　要：目的:基于"心受气于脾"探讨心痛泰调控p38丝裂原活化蛋白激酶(p38 MAPK)/转录激活因子-1(AP-1)对动脉粥样硬化兔的中医证候积分和血管平滑肌细胞(VSMC)胶原纤维的影响。方法:120只清洁级兔随机分为假手术组,痰瘀互结模型组,心痛泰低剂量组、中剂量组、高剂量组和瑞舒伐他汀组。采用高脂喂养+球囊损伤法,造成痰瘀互结病证结合动脉粥样硬化兔模型,造模后予以相应药物灌胃8周(心痛泰低、中、高剂量组和瑞舒伐他汀组的给药量分别为2.3,4.6,9.2 g·kg^(-1)和0.55 mg·kg^(-1))。给药周期结束时取腹主动脉,苏木素-伊红(HE)染色观察易损斑块的情况;免疫组化法(IHC)测定基质金属蛋白酶-9(MMP-9),组织基质金属蛋白酶抑制剂-1(TIMP-1);马松(Masson)染色观察平滑肌细胞胶原纤维分解情况;蛋白免疫印迹法(Western blot)测定主动脉组织p38 MAPK,AP-1蛋白的表达;采用中医证候评分表评价痰瘀互结证中医证候积分。结果:与模型组比较,心痛泰各剂量组、瑞舒伐他汀组MMP-9含量显著降低,TIMP-1含量显著升高,p38 MAPK蛋白的表达,AP-1的核转位显著降低(P<0.01);与心痛泰低剂量组比较,心痛泰中、高剂量组,瑞舒伐他汀组MMP-9明显降低,TIMP-1明显升高,p38 MAPK蛋白的表达,AP-1的核转位明显降低(P<0.05,P<0.01)。与模型组比较,心痛泰各剂量组、瑞舒伐他汀组的中医证候积分均明显改善(P<0.05,P<0.01);与心痛泰低剂量组比较,心痛泰中、高剂量组,瑞舒伐他汀组的中医证候积分均显著改善(P<0.01)。Masson染色显示,模型组平滑肌纤维排列紊乱,胶原分解增多,纤维帽变薄,斑块易损性增加;与模型组比较,心痛泰各组和瑞舒伐他汀组平滑肌细胞排列较为整齐,胶原纤维分解减少,斑块稳定性增加。结论:心痛泰可下调p38 MAPK,MMP-9的表达,提高TIMP-1的水平,减少AP-1核转位,减少平滑肌细胞胶原纤维的分解,改善痰瘀互结证的中医证�Objective: To explore the effects of Xintongtai(XTT)on traditional Chinese medicine(TCM) syndrome score and collagen fibers in vascular smooth muscle cells(VSMCs) of rabbits with atherosclerosis in the regulation of p38 mitogen-activated protein kinase(p38 MAPK)/activator protien-1(AP-1)signaling pathway. Method:A total of 120 rabbits of SPF grade were randomly divided into the sham operation group,combined phlegm and blood stasis model group,rosuvastatin group,and low-,middle-,and high-dose XTT groups. The rabbit model of atherosclerosis due to combined phlegm and blood stasis was established by exposing them to high-fat diet and balloon injury. Following modeling, the corresponding drugs were administered by gavage for eight weeks(2.3,4.6,9.2 g·kg^(-1) for low-,middle-,and high-dose XTT groups and0.55 mg·kg^(-1) for rosuvastatin group). At the end of medication,the abdominal aorta was isolated and stained with htoxylin-eosin(HE)for observing the vulnerable plaque. Matrix metalloproteinase-9(MMP-9)and tissue inhibitor of metalloproteinase-1(TIMP-1)were detected by immunohistochemistry(IHC). The collagen fiber decomposition in VSMCs was observed after Masson staining. The protein expression levels of p38 MAPK and AP-1 in aorta was assayed by Western blotting. The combined phlegm and blood stasis syndrome was scored based on TCM syndrome scoring scale. Result: Compared with the model group,XTT at each dose and rosuvastatin significantly decreased MMP-9 content,increased TIMP-1,down-regulated p38 MAPK protein expression,and weakened the nuclear translocation of AP-1(P<0.01). Compared with the low-dose XTT group,the middle-and high-dose XTT groups and rosuvastatin group exhibited obviously lowered MMP-9,elevated TIMP-1, down-regulated p38 MAPK protein expression, and diminished AP-1 nuclear translocation(P<0.05,P<0.01). The TCM syndrome scores of the middle-and high-dose XTT groups and rosuvastatin group were significantly improved as compared with that in the model group(P<0.05,P<0.01). The comparison with the lo

关 键 词：心受气于脾 心痛泰 动脉粥样硬化 p38丝裂原活化蛋白激酶(p38 MAPK)/转录激活因子-1(AP-1)信号通路 中医证候积分 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学]