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基于局部MCP-1/MMP-2/TGF-β1促炎信号环探讨温和灸对衰老大鼠血管衰老的影响

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

作者信息

Li Yi, Li Xue-Zhi, Liao Dong-Mei, Pang Fang, Yao Tai-Wan, He Xin-Lu

机构信息

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.

出版信息

Zhen Ci Yan Jiu. 2022 Apr 25;47(4):290-7. doi: 10.13702/j.1000-0607.20210472.

Abstract

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·d) 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·[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 (<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 (<0.01). After the interventions, the decreased levels of DES, serum TT and FT contents and the increased levels of serum AngⅡ, collagen fiber contents, aortic MCP-1 immunoactivity and MMP-2, TGF-β1 and MCP-1 protein expression were reversed in both medication and moxibustion groups (<0.01, <0.05). The therapeutic effect of mild moxibustion was significantly superior to that of medication in down-regulating the aortic collagen fiber, serum AngⅡ contents and MCP-1 immunoactivity and protein expression (<0.05). H.E. staining showed thickened endometrium and disordered arrangement of vascular smooth muscles of the aorta in the senium group, and thinner endometrium and regular and ordered arrangement of aortic vascular smooth muscles in both moxibustion and medication groups.

CONCLUSION

Mild moxibustion may improve vascular aging in senescence rats, which is possibly by suppressing vascular MCP-1/MMP-2/TGF-β1 pro-inflammatory signal loop.

摘要

目的

观察温和灸对衰老大鼠单核细胞趋化蛋白1(MCP-1)/基质金属蛋白酶2(MMP-2)/转化生长因子β1(TGF-β1)促炎信号环的影响,以探讨其改善血管衰老(VA)的机制。

方法

将24只雄性VA SD大鼠随机分为衰老(VA)对照组、药物组和艾灸组,另取8只2月龄的年轻SD大鼠作为年轻对照组。通过腹腔注射D-半乳糖(300 mg·kg·d),每日1次,连续4周建立VA模型,并通过血清总睾酮(TT)和游离睾酮(FT)水平进行验证。对艾灸组大鼠双侧“肾俞”(BL23)和“关元”(CV4)进行温和灸,每次20分钟,每天1次,每周5天,共8周。药物组大鼠除周末外,每日腹腔注射丙酸睾酮(7 mg·kg·[3 d]),连续8周;衰老对照组和年轻对照组大鼠除周末外,每日腹腔注射相同剂量的生理盐水,连续8周。记录治疗前后的力竭游泳时间(DES)。分别采用苏木精-伊红(H.E.)染色和Masson染色观察胸主动脉组织的组织病理学变化和胶原纤维含量。采用酶联免疫吸附测定(ELISA)法测定血清TT、FT和血管紧张素2(AngⅡ)的含量。采用免疫组织化学法检测主动脉MCP-1的免疫活性,采用蛋白质印迹法检测主动脉MCP-1、MMP-2和TGF-β1的表达水平。

结果

与年轻对照组相比,衰老对照组的DES、血清TT和FT含量显著降低(<0.01),而血清AngⅡ、胶原纤维含量、主动脉MCP-1免疫活性以及MMP-2、TGF-β1和MCP-1蛋白表达显著升高(<0.01)。干预后,药物组和艾灸组DES、血清TT和FT含量降低以及血清AngⅡ、胶原纤维含量、主动脉MCP-1免疫活性以及MMP-2、TGF-β1和MCP-1蛋白表达升高的情况均得到逆转(<0.01,<0.05)。在下调主动脉胶原纤维、血清AngⅡ含量以及MCP-1免疫活性和蛋白表达方面,温和灸的治疗效果显著优于药物组(<0.05)。H.E.染色显示,衰老组大鼠的主动脉内膜增厚,血管平滑肌排列紊乱;艾灸组和药物组大鼠的主动脉内膜变薄,血管平滑肌排列规则有序。

结论

温和灸可能通过抑制血管MCP-1/MMP-2/TGF-β1促炎信号环改善衰老大鼠的血管衰老。

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