基于MCP-1/MMP-2/TGF-β1局部促炎信号环探讨温和灸对老年大鼠血管衰老的干预作用  

Effect of mild moxibustion on vascular aging in senescence rats based on local MCP-1/MMP-2/TGF-β1 pro-inflammatory signal loop

作　　者：李怡 李学智[1] 廖冬梅 庞芳 姚太万 何昕璐 LI Yi;LI Xue-zhi;LIAO Dong-mei;PANG Fang;YAO Tai-wan;HE Xin-lu(College of Traditional Chinese Medicine,Chongqing Medical University,Chongqing Key Laboratory of Traditional Chinese Medicine for Prevention and Cure of Metabolic Diseases,Chongqing 400016,China;Department of Traditional Chinese Medicine,Chongqing General Hospital,Chongqing 400014)

机构地区：[1]重庆医科大学中医药学院中医药防治代谢性疾病重庆市重点实验室,重庆400016 [2]重庆市人民医院中医科,重庆400014

出　　处：《针刺研究》2022年第4期290-297,共8页Acupuncture Research

基　　金：重庆市基础科学与前沿技术专项项目(No.cstc2017jcyjAX0213)。

摘　　要：目的:观察温和灸对老年大鼠血管的抗老化作用,探讨温和灸通过调控单核细胞趋化蛋白1(MCP-1)/基质金属蛋白酶2(MMP-2)/转化生长因子β1(TGF-β1)局部促炎信号环对抗血管老化的机制。方法:50只8月龄雄性SD大鼠腹腔注射D-半乳糖300 mg·kg^(-1)·d^(-1),连续4周,制备衰老大鼠模型。24只造模成功的大鼠随机分为老年对照组、药物组和温和灸组,2月龄青年大鼠为青年对照组,每组8只。温和灸组选择“关元”及双侧“肾俞”进行温和灸治疗,1次/d, 20 min/次,治疗5 d休息2 d,持续治疗8周;药物组予丙酸睾酮注射液[7 mg·kg^(-1)·(3 d)^(-1)]腹腔注射,连续8周;老年对照组、青年对照组予0.9%氯化钠溶液腹腔注射,剂量与疗程同药物组。观察各组大鼠治疗前后游泳力竭时间;HE和Masson染色法观察各组大鼠胸主动脉形态和胶原纤维含量的变化;酶联免疫吸附测定法检测各组大鼠血清总睾酮(TT)、游离睾酮(FT)、血管紧张素2(AngⅡ)含量;免疫组织化学法检测大鼠胸主动脉中MCP-1阳性表达水平;Wes-tern blot法检测大鼠胸主动脉中MMP-2、TGF-β1、MCP-1蛋白表达量。结果:与青年对照组比较,老年对照组大鼠游泳力竭时间明显缩短(P<0.01),血清TT、FT含量明显降低(P<0.01),血清AngⅡ含量和胸主动脉胶原纤维含量、MCP-1阳性表达及MMP-2、TGF-β1、MCP-1的蛋白表达量明显升高(P<0.01)。治疗后与老年对照组比较,药物组、温和灸组游泳力竭时间明显延长(P<0.01),血清TT、FT含量明显升高(P<0.01),血清AngⅡ含量、胸主动脉胶原纤维含量、MCP-1阳性表达及MMP-2、TGF-β1、MCP-1的蛋白表达量明显降低(P<0.01,P<0.05)。与药物组比较,温和灸组血清AngⅡ含量、胸主动脉胶原纤维含量、MCP-1阳性表达均降低(P<0.05)。青年对照组大鼠主动脉内膜较薄,平滑肌细胞排列整齐;老年对照组内膜明显增厚,且平滑肌细胞排列紊乱;与老年对照组比较,药物组�Objective To observe the effect of mild moxibustion on monocyte chemotaxis protein 1(MCP-1)/matrix metalloproteinase 2(MMP-2)/transforming growth factor β1(TGF-β1) pro-inflammatory signal loop in senile rats, so as to explore its mechanisms underlying improving vascular aging(VA). Methods Twenty-four male VA SD rats were randomized into senium(VA) control, medication and moxibustion groups, and other 8 young SD rats(aged 2 months) were used as the young control group. The VA model was established by intraperitoneal injection of D-galactose(300 mg·kg^(-1)·d^(-1)) once daily for 4 weeks, and verified by serum total testosterone(TT) and free testosterone(FT) levels. For rats of the moxibustion group, mild moxibustion was applied to bilateral “Shenshu”(BL23) and “Guanyuan”(CV4) for 20 min, once a day, 5 days a week for 8 weeks. Rats of the medication group were treated by intraperitoneal injection of testosterone propionate(7 mg·kg^(-1)·[3 d];) once daily for 8 weeks except weekends, and rats of the senium control and young control groups treated by intraperitoneal injection of the same dose of normal saline, once daily for 8 weeks except weekends. The duration of exhausted swimming(DES) before and after the treatment was recorded. H.E. staining and Masson staining were used to observe histopathological changes and collagen fiber content of the thoracic aortic tissue, respectively. The contents of serum TT, FT and angiotensin 2(Ang Ⅱ) were determined by ELISA. The immunoactivity of aortic MCP-1 was detected by immunohistochemistry, and the expression levels of aortic MCP-1, MMP-2 and TGF-β1 were detected by Western blot. Results Compared with the young control group, the levels of DES, serum TT and FT contents were significantly decreased(P<0.01), while those of serum AngⅡ and collagen fiber contents, aortic MCP-1 immunoactivity and MMP-2, TGF-β1 and MCP-1 protein expression considerably increased in the senium control group(P<0.01). After the interventions, the decreased levels of DES, serum TT a

关 键 词：温和灸 睾酮 血管老化 衰老 

分 类 号：R245.81[医药卫生—针灸推拿学]