芪参益气滴丸通过TRPC1/STIM1通路调节Ca~(2+)稳态发挥抗动脉粥样硬化作用
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摘要
目的:探究芪参益气滴丸(Qishen-Yiqi dropping pills,QS)治疗动脉粥样硬化(artherosclerosis,AS)的作用机制。方法:利用高脂饮食建立SD大鼠AS模型,随机分为:正常对照组,模型组,芪参益气滴丸低、中、高剂量组,阳性对照组,每组6只。处理12周后收集血清检测各组大鼠血脂及Ca~(2+)水平;HE染色观察动脉组织形态学变化;ELISA法检测血清炎症因子白细胞介素1β(interleukin-1β,IL-1β)、IL-6和肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)水平;硝酸还原酶法检测动脉组织中一氧化氮(nitric oxide,NO)水平;Western blot检测动脉组织中瞬时受体电位通道蛋白1(transient receptor potential channel protein 1,TRPC1)、基质交互分子1(stromal interaction molecule 1,STIM1)和内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)蛋白表达水平。结果:芪参益气滴丸能减轻AS大鼠动脉内膜增厚及血管狭窄,抑制AS斑块形成;与模型组相比,芪参益气滴丸能显著降低大鼠血液中总胆固醇(TC)、甘油三酯(TG)和低密度脂蛋白胆固醇(LDL-C)水平,提高高密度脂蛋白胆固醇(HDL-C)水平(P<0.05);芪参益气滴丸治疗组大鼠血清炎症因子IL-1β、IL-6和TNF-α水平与AS大鼠相比显著降低(P<0.05);芪参益气滴丸治疗组大鼠血清Ca~(2+)水平显著低于且动脉组织中NO水平显著高于AS大鼠(P<0.05);与AS大鼠相比,芪参益气滴丸治疗组大鼠动脉组织中TRPC1和STIM1蛋白水平显著降低,且eNOS蛋白表达水平显著升高(P<0.05)。结论:芪参益气滴丸可以通过TRPC1/STIM1通路调节钙稳态,促进血管舒张因子NO的合成释放,抑制炎症反应,从而发挥抗AS作用。
AIM: To explore the therapeutic effect of Qishen-Yiqi dripping pills(QS) on atherosclerosis(AS) and the mechanism. METHODS: AS rat model was established by high-fat diet, and SD rats were randomly divided into normal control group, AS model group, low-dose, middle-dose and high-dose QS groups, and positive group(n=6 each). After administration for 12 weeks, serum samples were collected to detect the serum lipid and Ca~(2+) levels. HE staining was used evaluated the histopathological changes of arterial tissue. The serum levels of interleukin-1β(IL-1β), IL-6 and tumor necrosis factor-α(TNF-α) were measured by ELISA. The nitric oxide(NO) level was detected by nitrate reductase method. The protein levels of transient receptor potential channel protein 1(TRPC1), stromal interaction molecule 1(STIM1) and endothelial NO synthase(eNOS) were determined by Western blot. RESULTS: QS significantly reduced the arterial damage via inhibiting the formation of atherosclerotic plaque and attenuated intimal thickening and vascular stenosis. Compared with AS group, the serum levels of total cholesterol(TC), triglyceride(TG) and low-density lipoprotein cholesterol(LDL-C) were decreased significantly and the levels of high-density lipoprotein cholesterol(HDL-C) were increased significantly in high-dose QS group(P<0.05). The serum levels of IL-1β, IL-6 and TNF-α in high-dose QS group were lower than those in AS group(P<0.05). Compared with AS group, the serum Ca~(2+) level was lowered and the arterial tissue NO level was elevated in QS groups(P<0.05). Compared with AS rats, the protein levels of TRPC1 and STIM1 were decreased significantly and the protein level of eNOS was increased significantly in the rats treated with QS(P<0.05). CONCLUSION: QS regulate calcium homeostasis via TRPC1/STIM1 pathway, increase the production of NO and inhibit the inflammatory responses, thus exerting anti-AS effect.
AIM: To explore the therapeutic effect of Qishen-Yiqi dripping pills(QS) on atherosclerosis(AS) and the mechanism. METHODS: AS rat model was established by high-fat diet, and SD rats were randomly divided into normal control group, AS model group, low-dose, middle-dose and high-dose QS groups, and positive group(n=6 each). After administration for 12 weeks, serum samples were collected to detect the serum lipid and Ca~(2+) levels. HE staining was used evaluated the histopathological changes of arterial tissue. The serum levels of interleukin-1β(IL-1β), IL-6 and tumor necrosis factor-α(TNF-α) were measured by ELISA. The nitric oxide(NO) level was detected by nitrate reductase method. The protein levels of transient receptor potential channel protein 1(TRPC1), stromal interaction molecule 1(STIM1) and endothelial NO synthase(eNOS) were determined by Western blot. RESULTS: QS significantly reduced the arterial damage via inhibiting the formation of atherosclerotic plaque and attenuated intimal thickening and vascular stenosis. Compared with AS group, the serum levels of total cholesterol(TC), triglyceride(TG) and low-density lipoprotein cholesterol(LDL-C) were decreased significantly and the levels of high-density lipoprotein cholesterol(HDL-C) were increased significantly in high-dose QS group(P<0.05). The serum levels of IL-1β, IL-6 and TNF-α in high-dose QS group were lower than those in AS group(P<0.05). Compared with AS group, the serum Ca~(2+) level was lowered and the arterial tissue NO level was elevated in QS groups(P<0.05). Compared with AS rats, the protein levels of TRPC1 and STIM1 were decreased significantly and the protein level of eNOS was increased significantly in the rats treated with QS(P<0.05). CONCLUSION: QS regulate calcium homeostasis via TRPC1/STIM1 pathway, increase the production of NO and inhibit the inflammatory responses, thus exerting anti-AS effect.
引文
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[2] Rosales C, Davidson WS, Gillard BK, et al. Speciated high-density lipoprotein biogenesis and functionality[J]. Curr Atheroscler Rep, 2016, 18(5):25.
[3] Nahrendorf M, Swirski FK. Lifestyle effects on hematopoiesis and atherosclerosis[J]. Circ Res, 2015, 116(5):884-894.
[4] Daiber A, Steven S, Weber A, et al. Targeting vascular (endothelial) dysfunction[J]. Br J Pharmacol, 2017, 174(12):1591-1619.
[5] Rattanakul C, Lenbury Y. Cellular automata simulation of signal transduction and calcium dynamics with healthy and faulty receptor trafficking[C]// IEEE. Annual Systems Conference. Orlando: IEEE, 2016:147-154.
[6] Maehara A, Stone GW. High-risk coronary atherosclerosis: is it the plaque burden, the calcium, the lipid, or something else?[J]. Circ Cardiovasc Imaging, 2017, 10(10):e007116.
[7] Lewis RS. The molecular choreography of a store-operated calcium channel[J]. Nature, 2007, 446(7133):284-287.
[8] Shoenfeld Y, Sherer Y, Harats D. Atherosclerosis as an infectious, inflammatory and autoimmune disease[J]. Trends Immunol, 2001, 22(6):293-295.
[9] Kawashima S, Yokoyama M. Dysfunction of endothelial nitric oxide synthase and atherosclerosis[J]. Arterioscler Thromb Vasc Biol, 2004, 24(6):998-1005.
[10] 李欲来, 张军平, 赵广荣, 等. 芪参益气滴丸对气虚血瘀型冠心病患者血脂影响观察研究[J]. 中华实用中西医杂志, 2005, 18(10):1421-1423.
[11] 宋郁珍, 郭利平, 商洪才, 等. 芪参益气滴丸对实验性高胆固醇血症家兔脂代谢的影响[J]. 吉林中医药, 2011, 31(1):71-73.
[12] 缪静, 陈洁, 周鑫斌, 等. 丹蒌片对动脉粥样硬化模型大鼠抵抗素和血管内皮的影响[J]. 新中医, 2016, (4):269-272.
[13] Dinh QN, Chrissobolis S, Diep H, et al. Advanced atherosclerosis is associated with inflammation, vascular dysfunction and oxidative stress, but not hypertension[J]. Pharmacol Res, 2016, 116:70-76.
[14] Nonogaki K, Fuller GM, Fuentes NL, et al. Interleukin-6 stimulates hepatic triglyceride secretion in rats[J]. Endocrinology, 1995, 136(5):2143-2149.
[15] Rizvi AA. Inflammation markers as mediators of vasculo-endothelial dysfunction and atherosclerosis in the metabo-lic syndrome and type 2 diabetes[J]. Chin Med J (Engl), 2007, 120(21):1918-1924.
[16] Popa C, Netea MG, van Riel PL, et al. The role of TNF-α in chronic inflammatory conditions, intermediary metabolism, and cardiovascular risk[J]. J Lipid Res, 2007, 48(4):751-762.
[17] Rooney MR, Pankow JS, Sibley SD, et al. Serum calcium and incident type 2 diabetes: the Atherosclerosis Risk in Communities (ARIC) study[J]. Am J Clin Nutr, 2016, 104(4):1023-1029.
[18] Liu J, Xin L, Benson VL, et al. Store-operated calcium entry and the localization of STIM1 and Orai1 proteins in isolated mouse sinoatrial node cells[J]. Front Physiol, 2015, 6:69.
[19] Wang Y, Wang Y, Li GR. TRPC1/TRPC3 channels mediate lysophosphatidylcholine-induced apoptosis in cultured human coronary artery smooth muscles cells[J]. Oncotarget, 2016, 7(32):50937-50951.
[20] 李永胜, 王照华, 梁黔生,等. 丹参酮ⅡA对血管紧张素Ⅱ所致主动脉内皮细胞游离钙离子及产生一氧化氮的影响[J]. 中华高血压杂志, 2006, 14(11):882-886.