天麻钩藤饮对自发性高血压大鼠p38 MAPK/HSP27通路影响的研究
Study on the effect of
Tianma Gouteng Decoction on the p38 MAPK/HSP27 pathway in spontaneously hypertensive rats- 北京中医药大学学报 2023年46卷第12期 页码：1694-1705
- 作者机构：
1.中南大学湘雅医院国际医疗部 长沙 410008
2.中南大学湘雅医院老年医学科
3.中南大学湘雅医院中西医结合科
- 作者简介：
秦钦，女，在读硕士生
#钟广伟，男，博士，副教授，硕士生导师，主要研究方向：中医药衰老理论及高血压防治，Email：zgw7512@sina.com
- 基金信息：国家自然科学基金项目(81673858);湖南省自然科学基金项目(2023JJ30979;2021JJ31118);湖南省中医药科研计划项目(C2022032)
DOI：10.3969/j.issn.1006-2157.2023.12.010
中图分类号：
扫 描 看 全 文
秦钦, 时拥月, 姚佳梅, 等. 天麻钩藤饮对自发性高血压大鼠p38 MAPK/HSP27通路影响的研究[J]. 北京中医药大学学报, 2023,46(12):1694-1705.
QIN Qin, SHI Yongyue, YAO Jiamei, et al. Study on the effect of
Tianma Gouteng Decoction on the p38 MAPK/HSP27 pathway in spontaneously hypertensive rats[J]. Journal of Beijing University of Traditional Chinese Medicine, 2023,46(12):1694-1705.秦钦, 时拥月, 姚佳梅, 等. 天麻钩藤饮对自发性高血压大鼠p38 MAPK/HSP27通路影响的研究[J]. 北京中医药大学学报, 2023,46(12):1694-1705. DOI： 10.3969/j.issn.1006-2157.2023.12.010.
QIN Qin, SHI Yongyue, YAO Jiamei, et al. Study on the effect of
Tianma Gouteng Decoction on the p38 MAPK/HSP27 pathway in spontaneously hypertensive rats[J]. Journal of Beijing University of Traditional Chinese Medicine, 2023,46(12):1694-1705. DOI： 10.3969/j.issn.1006-2157.2023.12.010.
摘要
目的,2,探讨p38丝裂原活化蛋白激酶(p38 MAPK)/热休克蛋白27(HSP27)通路在天麻钩藤饮干预自发性高血压大鼠(SHR)中的作用机制。,方法,2,将30只SHR按随机数字表法分为模型组、缬沙坦组(0.03 g/kg)和天麻钩藤饮组(1 g/kg)，每组各10只，WKY大鼠10只为正常组，缬沙坦组和天麻钩藤饮组大鼠分别给予相应药物灌胃，正常组与模型组大鼠灌胃等量生理盐水，连续8周。测量第0、14、28、42、56天各组大鼠的血压；给药结束后，观察各组大鼠的行为学变化；采用苏木精－伊红染色法观察血管形态结构变化，天狼星红染色法观察血管纤维化程度；采用电子显微镜观察细胞自噬超微结构；采用免疫荧光法检测血管组织中磷酸化p38 MAPK(p-p38 MAPK)和磷酸化HSP27(p-HSP27)的平均荧光强度；采用分光光度法检测血浆丙二醛(MDA)、超氧化物歧化酶(SOD)和血管组织抗超氧阴离子的含量；采用β-半乳糖苷酶(SA-β-gal)染色法观察血管衰老改变；采用实时荧光PCR法检测血管组织中p38 MAPK、MAP激酶活化蛋白激酶－2(MAPKAPK-2)、HSP27、自噬相关蛋白5(ATG5)及p62 mRNA表达；采用蛋白质印迹法检测血管组织中p-p38 MAPK、磷酸化MAPKAPK-2(p-MAPKAPK-2)、p-HSP27、ATG5及p62蛋白表达。,结果,2,与正常组比较，模型组大鼠血压和易激惹程度升高，旋转耐受时间缩短；血管中膜厚度(MT)和MT/血管内径(LD)的值增加，自噬小体数量减少，SA-β-gal阳性细胞率升高，MDA含量升高、SOD含量和抗超氧阴离子含量降低；p-p38 MAPK平均荧光强度升高，p-p38 MAPK、p-MAPKAPK-2蛋白及p62 mRNA和蛋白表达升高，p-HSP27平均荧光强度减少，HSP27、ATG5 mRNA和p-HSP27、ATG5蛋白表达下降(均,P,<,0.05)。与模型组比较，缬沙坦组和天麻钩藤饮组大鼠血压和易激惹程度下降、旋转耐受时间延长；MT、MT/LD降低，自噬小体数量增多，SA-β-gal阳性细胞率减少，MDA含量降低、SOD含量和抗超氧阴离子含量升高；p-p38 MAPK平均荧光强度下降，p-p38 MAPK、p-MAPKAPK-2蛋白及p62 mRNA和蛋白表达降低，p-HSPA27平均荧光强度升高，HSP27、ATG5 mRNA和p-HSP27、ATG5蛋白表达升高(均,P,<,0.05)。,结论,2,天麻钩藤饮能够延缓SHR的血管衰老，其机制可能是通过p38 MAPK/HSP27通路调节细胞自噬实现的。
Abstract
Objective,2,We aimed to investigate the mechanism of action of ,Tianma Gouteng, Decoction in the treatment of spontaneously hypertensive rats(SHR) by studying the p38 mitogen-activated protein kinase (p38 MAPK)/heat shock protein 27 (HSP27) pathway.,Methods,2,Using the random number table method, 30 SHR were divided into the model group, the Valsartan group (0.03 g/kg), and the ,Tianma Gouteng, Decoction group (1 g/kg), 10 rats per group. Additionally, 10 Wistar-Kyoto (WKY) rats were assigned to the normal group. Rats in the Valsartan group and the ,Tianma Gouteng, Decoction group were given corresponding drugs, while rats in the normal group and the model group were administered an equivalent amount of saline orally for 8 consecutive weeks. Blood pressure was recorded on days 0, 14, 28, 42, and 56. Following treatment completion, behavioral changes were observed. Hematoxylin-eosin staining was used to observe the changes in vascular morphology and structure. Masson staining was used to observe the degree of vascular fibrosis. The autophagy ultrastrucure were examined by electron microscopy. An immunofluorescence assay was conducted and the average fluorescence intensities of phosphorylated p38 MAPK (p-p38 MAPK) and phosphorylated HSP27 (p-HSP27) in vascular tissue were calculated. Plasma malondialdehyde (MDA) content, plasma superoxide dismutase (SOD) content, and vascular tissue anti-superoxide anion content were measured by spectrophotometry. Vascular aging was evaluated through a senescence-associated β-galactosidase (SA-β-gal) staining method. The mRNA expression levels of p38 MAPK, MAPK-activated protein kinase-2 (MAPKAPK-2), HSP27, autophagy-associated protein 5 (ATG5), and p62 in vascular tissue were determined by real-time PCR. The protein expression levels of p-p38 MAPK, p-MAPKAPK-2, p-HSP27, ATG5, and p62 in vascular tissue were determined by Western blotting.,Results,2,Compared with the normal group, the blood pressure and irritability were increased, the rotational tolerance time was decreased, the thickness of vascular media (MT) and the ratio of vascular media thickness to vascular inner diameter (MT/LD) were significantly increased, the number of autophagosomes was decreased, the proportion of SA-β-gal positive cells was increased, the content of MDA was increased, the content of SOD and anti-superoxide anion were decreased, the average fluorescence intensity of p-p38 MAPK was increased, the protein expression levels of p-p38 MAPK and p-MAPKAPK-2 were increased, the mRNA and protein levels of p62 were increased, the average fluorescence intensity of p-HSP27 was decreased, and the mRNA expression level of HSP27 and the protein expression levels of p-HSP27, and the mRNA and protein levels of ATG5 were decreased in the model group (,P,<,0.05). Compared with the model group, the blood pressure and irritability were decreased, the rotational tolerance time was increased, MT and MT/LD were decreased, the number of autophagosomes was increased, the proportion of SA-β-gal positive cells was decreased, the content of MDA was decreased, the content of SOD and anti-superoxide anion were increased, the average fluorescence intensity of p-p38 MAPK was decreased, the protein levels of p-p38 MAPK and p-MAPKAPK-2 were decreased, the mRNA and protein levels of p62 were decreased, and the average fluorescence intensity of p-HSP27, the mRNA expression level of HSP27 and the protein expression levels of p-HSP27, and the mRNA and protein expression levels of ATG5 were increased in the Valsartan group and the ,Tianma Gouteng, Decoction group (,P,<,0.05).,Conclusion,2,Tianma Gouteng, Decoction can delay vascular aging in a spontaneous hypertensive rat model, possibly by regulating autophagy through the p38 MAPK/HSP27 pathway.
关键词
天麻钩藤饮高血压p38丝裂原活化蛋白激酶/热休克蛋白27信号通路细胞自噬大鼠
Keywords
Tianma Gouteng Decoctionhypertensionp38 mitogen-activated protein kinase/heat shock protein 27 pathwayautophagyrats
references
MOTURI S，GHOSH-CHOUDHARY SK，FINKEL T. Cardiovascular disease and the biology of aging[J]. J Mol Cell Cardiol，2022，167：109－117.
CAO QY，LI M，WANG TG，et al. Association of early and supernormal vascular aging categories with cardiovascular disease in the Chinese population[J/OL]. Front Cardiovasc Med，2022，9：895792[2023－03－31]. https://www.frontiersin.org/articles/10.3389/fcvm.2022.895792https://www.frontiersin.org/articles/10.3389/fcvm.2022.895792.
JOHNSON GL，LAPADAT R. Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases[J]. Science，2002，298(54)：1911－1912.
PENG Y，PAN J，OU FT，et al. Fenbendazole and its synthetic analog interfere with HeLa cells' proliferation and energy metabolism via inducing oxidative stress and modulating MEK3/6－p38－MAPK pathway[J/OL]. Chem Biol Interact，2022，361：109983[2023－03－31]. https://www.sciencedirect.com/science/article/abs/pii/109983https://www.sciencedirect.com/science/article/abs/pii/109983.
WANG ZY，LI A，HUANG X，et al. HSP27 protects skin from ultraviolet B －induced photodamage by regulating autophagy and reactive oxygen species production[J/OL]. Front Cell Dev Biol，2022，10：852244[2023－03－31]. https://www.frontiersin.org/articles/10.3389/fcell.2022.852244https://www.frontiersin.org/articles/10.3389/fcell.2022.852244.
姚佳梅，张翠，杨玉书，等. 平肝潜阳法对高血压病患者血管衰老及miRNAs-HSP27表达的影响[J]. 中华中医药杂志，2021, 36(11)：6866－6873.
LI T, WU YN, WANG H, et al. Dapk1 improves inflammation, oxidative stress and autophagy in LPS-induced acute lung injury via p38 MAPK/NF－κB signaling pathway[J/OL]. Mol Immunol, 2020, 120：13－22[2023－03－31].https://www.sciencedirect.com/science/article/abs/pii/S0161589019307825https://www.sciencedirect.com/science/article/abs/pii/S0161589019307825.
李湘平，罗艳红，钟广伟，等. 平肝潜阳方含药血清中天麻素水平测定及其对血管平滑肌细胞活性的影响[J]. 中国组织工程研究与临床康复，2010，14(15)：2735－2739.
陈奇. 中药药理研究方法学[M].3版．北京：人民卫生出版社，2011：1261－1263.
孙明祎. 天麻钩藤饮对原发性高血压患者血压变异性及血管内皮保护机制作用研究[J]. 长春中医药大学学报，2019，35(2)：261－263.
李云攀，刘美云. 天麻钩藤饮对自发性高血压大鼠血管内皮细胞自噬相关信号通路Ca2＋/AMPK/mTOR的影响[J]. 中国现代医学杂志，2018，28(22)：38－43.
BRUNO RM，DURANTI E，IPPOLITO C，et al. Different impact of essential hypertension on structural and functional age-related vascular changes[J]. Hypertension，2017，69(1)：71－78.
STABENOW LK，ZIBROVA D，ENDER C，et al. Oxidative glucose metabolism promotes senescence in vascular endothelial cells[J/OL]. Cells，2022，11：2213[2023－03－31].https://www.mdpi.com/2073-4409/11/14/2213https://www.mdpi.com/2073-4409/11/14/2213.
陈波，陆燕蓉，陈又南，等. 血管内皮衰老和氧化应激[J]. 生理科学进展，2015，46(1)：23－27.
YU N，SHEN A，CHU JF，et al. Qingda granule inhibits angiotensin Ⅱ induced VSMCs proliferation through MAPK and PI3K/AKT pathways[J/OL]. J Ethnopharmacol，2020，258：112767[2023－03－31]. https://www.sciencedirect.com/science/article/abs/pii/112767https://www.sciencedirect.com/science/article/abs/pii/112767.
TANIYAMA Y，USHIO-FUKAI M，HITOMI H，et al. Role of p38 MAPK and MAPKAPK－2 in angiotensin II-induced Akt activation in vascular smooth muscle cells[J]. Am J Physiol Cell Physiol，2004，287(2)：494－499.
姚佳梅，杨海燕，杨玉书，等．天麻钩藤饮调控MFN2表达干预自发性高血压病模型大鼠血管衰老的作用机制[J].中国中药杂志，2022，47(14)：3844－3852.
PARK AM，TSUNODA I，YOSHIE O. Heat shock protein 27 promotes cell cycle progression by down-regulating E2F transcription factor 4 and retinoblastoma family protein p130[J]. J Biol Chem，2018，293(41)：15815－15826.
CHEN SF，KANG ML，CHEN YC，et al. Autophagy-related gene 7 is downstream of heat shock protein 27 in the regulation of eye morphology, polyglutamine toxicity, and lifespan in Drosophila[J/OL]. J Biomed Sci，2012，19：52[2023－03－31]. http://www.jbiomedsci.com/content/19/1/52http://www.jbiomedsci.com/content/19/1/52.
ZHANG L，ZHOU J，JING ZP，et al. Glucocorticoids regulate the vascular remodeling of aortic dissection via the p38 MAPK-HSP27 pathway mediated by soluble TNF-RII[J]. EBioMedicine，2018，27(1)：247－257.
LEVINE B，KROEMER G. Biological functions of autophagy genes: A disease perspective[J]. Cell，2019，176(1－2)：11－42.
LUO ZL，XU WH，MA S，et al. Moderate autophagy inhibits vascular smooth muscle cell senescence to stabilize progressed atherosclerotic plaque via the mTORC1/ULK1/ATG13 signal pathway[J/OL]. Oxid Med Cell Longev，2017，2017：3018190[2023－03－31]. https://www.hindawi.com/journals/omcl/2017/3018190https://www.hindawi.com/journals/omcl/2017/3018190.
MONTEZANO AC，TOUYZ RM. Reactive oxygen species, vascular Noxs, and hypertension: focus on translational and clinical research[J].Antioxid Redox Signal，2014，20(1)：164－182.
陶丽丽，雷燕. 人参、三七、川芎提取物对血管平滑肌细胞衰老相关β半乳糖苷酶及p16－cyclinD/CDK-Rb通路的影响[J]. 中西医结合学报，2012，10(1)：76－84.
KIM EN，KIM MY，LIM JH，et al. The protective effect of resveratrol on vascular aging by modulation of the renin-angiotensin system[J].Atherosclerosis，2018，270(1)：123－131.
0
浏览量
0
下载量
0
CSCD
- BibTeX下载
- Endnote下载
- RefMan 下载
- NoteExpress下载
- NoteFirst下载
关联资源
相关文章
相关作者
相关机构
- 地址：北京市朝阳区北三环东路11号 邮编：100029
- 联系电话：(010)53911857 Email：jbutcm@bucm.edu.cn
- 技术支持由北京北大方正电子有限公司提供 京ICP备09064830号-19
京公网安备11010802024621 - 本系统建议在Chrome、 IE9+ 以上版本浏览器阅读本站内容，360浏览器请切换至极速模式
- Cookies帮助我们提供服务并提供个性化体验。使用本网站，即表示您同意我们使用Cookies