Influence of Yiqi Shengqing recipe on neuron pyroptosis in ischemic stroke rats by regulating HIF-1α/NLRP3 signal pathway

doi:10.11958/20231141

Abstract

Abstract:

Objective To investigate the influence of Yiqi Shengqing recipe on neuronal pyroptosis in ischemic stroke rats by regulating hypoxia inducible factor-1α (HIF-1α)/NOD-like receptor thermal protein domain associated protein 3 (NLRP3) signal pathway. Methods SD rats were randomly divided into the sham operation group (S group), the model group (M group), the low, medium and high dose (3.465, 6.930 and 13.860 g/kg) Yiqi Shengqing recipe groups, and high-dose Yiqi Shengqing recipe+HIF-1α activator DMOG (13.860 g/kg Yiqi Shengqing recipe+40 mg/kg DMOG) group, 15 animals in each group. The stroke model was constructed by thread plug method. After successful modeling, neural function defects were evaluated including TTC staining for evaluating the volume of cerebral infarction, ELISA for detecting the serum interleukin (IL)-1β and IL-18 levels, HE staining for detecting pathological changes of ischemic cortex, TUNEL staining for detecting neuronal apoptosis and Western blot assay for detecting the expression of pyroprotein and HIF-1α/NLRP3 pathway protein. Results Compared with the S group, the neurological deficit score, infarct volume, IL-1β, IL-18 contents, apoptosis rate of nerve cells, GasderminD-N (GSDMD-N), Caspase-1, HIF-1α and NLRP3 protein levels were increased in the M group (P<0.05). Compared with the M group, the neurological defect score, infarct volume, IL-1β, IL-18 contents, neuronal apoptosis rate, GSDMD-N, Caspase-1, HIF-1α and NLRP3 protein levels were decreased in the low, medium and high dose Yiqi Shengqing recipe groups (P<0.05). DMOG attenuated the improvement effect of high-dose Yiqi Shengqing recipe on neuronal death in ischemic stroke rats. Conclusion Yiqi Shengqing recipe may reduce the neuronal pyroptosis in ischemic stroke rats by down-regulating HIF-1α/NLRP3 signal pathway.

Key words: stroke, hypoxia-inducible factor 1, alpha subunit, NLR family, pyrin domain-containing 3 protein, neurons, pyroptosis, Yiqi Shengqing recipe

CLC Number:

R285.5

Figures/Tables 9

References 20

[1]	樊泽新, 刘广志. 缺血性卒中免疫学研究进展[J]. 中国现代神经疾病杂志, 2022, 22(1):46-53.
	FAN Z X, LIU G Z. Advances in immunology of ischemic stroke[J]. Chin J Contemp Neurol Neurosurg, 2022, 22(1):46-53. doi:10.3969/j.issn.1672-6731.2022.01.007.
[2]	GONCALVES A, SU E J, MUTHUSAMY A, et al. Thrombolytic tPA-induced hemorrhagic transformation of ischemic stroke is mediated by PKCβ phosphorylation of occludin[J]. Blood, 2022, 140(4):388-400. doi:10.1182/blood.2021014958.
[3]	QIU L, FU F, ZHANG W, et al. Prevalence, risk factors, and clinical outcomes of remote intracerebral hemorrhage after intravenous thrombolysis in acute ischemic stroke:a systematic review and meta-analysis[J]. J Neurol, 2023, 270(2):651-661. doi:10.1007/s00415-022-11414-2.
[4]	WU X, WEI J, YI Y, et al. Epimedium aqueous extract ameliorates cerebral ischemia/reperfusion injury through inhibiting ROS/NLRP3-mediated pyroptosis[J]. Antioxidants (Basel), 2023, 12(5):999. doi:10.3390/antiox12050999.
[5]	ABDEL-LATIF R G, RIFAAI R A, AMIN E F. Empagliflozin alleviates neuronal apoptosis induced by cerebral ischemia/reperfusion injury through HIF-1α/VEGF signaling pathway[J]. Arch Pharm Res, 2020, 43(5):514-525. doi:10.1007/s12272-020-01237-y.
[6]	JIANG Q, GENG X, WARREN J, et al. Hypoxia inducible factor-1α (HIF-1α) mediates NLRP3 inflammasome-dependent-pyroptotic and apoptotic cell death following ischemic stroke[J]. Neuroscience, 2020, 448:126-139. doi:10.1016/j.neuroscience.2020.09.036.
[7]	熊艳萍. 加味化痰通络汤结合针刺辅治脑卒中风痰瘀阻证疗效观察[J]. 实用中医药杂志, 2022, 38(11):1906-1907.
	XIONG Y P. Observation on the curative effect of Supplemented Huatan Tongluo Decoction combined with acupuncture in the treatment of wind-phlegm-stasis syndrome of cerebral apoplexy[J]. Journal of Practical Chinese Medicine, 2022, 38(11):1906-1907.
[8]	胡建芳, 余志辉, 陈朝俊, 等. 针刺治疗联合益气升清方对卒中后吞咽障碍患者预后的影响[J]. 中国实用医药, 2014, 9(21):167-168.
	HU J F, YU Z H, CHEN C J, et al. Effect of acupuncture treatment combined with Yiqi Shengqing Prescription on prognosis of patients with dysphagia after stroke[J]. China Prac Med, 2014, 9(21):167-168. doi:10.14163/j.cnki.11-5547/r.2014.21.009.
[9]	江丹, 汪何, 姚楠, 等. 基于TGF-β1/p38MAPK通路探讨益气升清方对糖尿病大鼠肾组织的影响[J]. 世界中医药, 2020, 15(16):2381-2386.
	JIANG D, WANG H, YAO N, et al. The effects of Yiqi Shengqing fomula on TGF-β1/p38MAPK signaling pathway in renal tissue of diabetes rats[J]. World Chinese Medicine, 2019, 15(16):2381-2386. doi:10.3969/j.issn.1673-7202.2020.16.006.
[10]	张佩, 贺建东, 韩冲芳, 等. HIF-1α/EphrinA1信号通路在糖尿病因素削弱七氟醚心肌保护中的作用机制[J]. 山西医科大学学报, 2021, 52(8):995-1000.
	ZHANG P, HE J D, HAN C F, et al. The mechanism of HIF-1α/EphrinA1 signaling pathway in diabetic impaired myocardial protection of sevoflurane[J]. J Shanxi Med Univ, 2021, 52(8):995-1000. doi:10.13753/j.issn.1007-6611.2021.08.010.
[11]	CHEN X, YANG T, LUO Y, et al. Methodological and reporting quality evaluation of Buyang Huanwu decoction for experimental cerebral ischemia-reperfusion injury:a systematic review[J]. Naunyn Schmiedebergs Arch Pharmacol, 2023, 396(5):831-849. doi:10.1007/s00210-022-02362-9.
[12]	单萍, 姚汉云, 王莉, 等. 基于网络药理学和分子对接的苏合香丸治疗缺血性卒中的机制探究[J]. 药物评价研究, 2022, 45(7):1294-1311.
	SHAN P, YAO H Y, WANG L, et al. Mechanism of Suhexiang pill in treatment of ischemic stroke based on network pharmacology and molecular docking[J]. Drug Evaluation Research, 2022, 45(7):1294-1311. doi:10.7501/j.issn.1674-6376.2022.07.011.
[13]	WANG L, LIU C, WANG L, et al. Astragaloside IV mitigates cerebral ischaemia-reperfusion injury via inhibition of P62/Keap1/Nrf2 pathway-mediated ferroptosis[J]. Eur J Pharmacol, 2023, 944:175516. doi:10.1016/j.ejphar.2023.175516.
[14]	付劭静, 周延华, 卑红喆, 等. 升麻素苷对脑缺血/再灌注大鼠脑损伤的保护作用及其机制研究[J]. 中国临床神经科学, 2021, 29(6):617-624.
	FU S J, ZHOU Y H, BEI H Z, et al. Protective effect and mechanism of Cimicifugoside from brain injury in rats undergoing cerebral ischemia reperfusion[J]. Chin J Clin Neurosci, 2021, 29(6):617-624.
[15]	伍忠根, 刘培文, 刘晖, 等. 葛根素对大鼠脑缺血-再灌注损伤的影响[J]. 江苏医药, 2021, 47(2):109-113.
	WU Z G, LIU P W, LIU H, et al. Effects of puerarin on cerebral ischemia-reperfusion injury in rats[J]. Jiangsu Med J, 2021, 47(2):109-113. doi:10.19460/j.cnki.0253-3685.2021.02.001.
[16]	薛丽, 韩红, 张力. 桑黄素通过抑制TXNIP/NLRP3/Caspase-1信号通路对脑缺血再灌注大鼠神经元凋亡的影响[J]. 天津医药, 2023, 51(5):487-491.
	XUE L, HAN H, ZHANG L. Effects of Morin on neuronal apoptosis in cerebral ischemia-reperfusion rats by inhibiting TXNIP/NLRP3/Caspase-1 signaling pathway[J]. Tianjin Med J, 2023, 51(5):487-491. doi:10.11958/20221258.
[17]	CAI L, YAO Z Y, YANG L, et al. Mechanism of electroacupuncture against cerebral ischemia-reperfusion injury:reducing inflammatory response and cell pyroptosis by inhibiting NLRP3 and Caspase-1[J]. Front Mol Neurosci, 2022, 15:822088. doi:10.3389/fnmol.2022.822088.
[18]	COLL R C, SCHRODER K, PELEGRÍN P. NLRP3 and pyroptosis blockers for treating inflammatory diseases[J]. Trends Pharmacol Sci, 2022, 43(8):653-668. doi:10.1016/j.tips.2022.04.003.
[19]	TSAO C C, BAUMANN J, HUANG S F, et al. Pericyte hypoxia-inducible factor-1 (HIF-1)drives blood-brain barrier disruption and impacts acute ischemic stroke outcome[J]. Angiogenesis, 2021, 24(4):823-842. doi:10.1007/s10456-021-09796-4.
[20]	CHEN S F, PAN M X, TANG J C, et al. Arginine is neuroprotective through suppressing HIF-1α/LDHA-mediated inflammatory response after cerebral ischemia/reperfusion injury[J]. Mol Brain, 2020, 13(1):63. doi:10.1186/s13041-020-00601-9.

组别	神经功能缺陷评分/分（n=15）	梗死体积/% （n=5）
S组	0.00±0.00	0.00±0.00
M组	2.93±0.26^a	17.03±1.70^a
L-Med组	2.13±0.35^b	12.87±1.45^b
M-Med组	1.20±0.41^bc	8.97±0.83^bc
H-Med组	0.47±0.52^bcd	5.00±0.05^bcd
H-Med+DMOG组	2.13±0.35^e	13.21±1.43^e
F	90.734^＊＊	67.926^＊＊

组别	神经功能缺陷评分/分（n=15）	梗死体积/% （n=5）
S组	0.00±0.00	0.00±0.00
M组	2.93±0.26^a	17.03±1.70^a
L-Med组	2.13±0.35^b	12.87±1.45^b
M-Med组	1.20±0.41^bc	8.97±0.83^bc
H-Med组	0.47±0.52^bcd	5.00±0.05^bcd
H-Med+DMOG组	2.13±0.35^e	13.21±1.43^e
F	90.734^＊＊	67.926^＊＊

组别	IL-1β	IL-18
S组	12.34±1.34	22.22±2.88
M组	50.76±5.56^a	57.43±5.95^a
L-Med组	42.45±4.65^b	46.31±4.62^b
M-Med组	30.34±3.26^bc	35.43±3.54^bc
H-Med组	21.23±2.17^bcd	24.00±3.13^bcd
H-Med+DMOG组	43.64±4.15^e	49.24±4.22^e
F	225.615^＊＊	174.083^＊＊

组别	IL-1β	IL-18
S组	12.34±1.34	22.22±2.88
M组	50.76±5.56^a	57.43±5.95^a
L-Med组	42.45±4.65^b	46.31±4.62^b
M-Med组	30.34±3.26^bc	35.43±3.54^bc
H-Med组	21.23±2.17^bcd	24.00±3.13^bcd
H-Med+DMOG组	43.64±4.15^e	49.24±4.22^e
F	225.615^＊＊	174.083^＊＊

组别	GSDMD-N/GAPDH	Caspase-1/GAPDH
S组	0.53±0.05	0.93±0.06
M组	1.35±0.16^a	2.65±0.28^a
L-Med组	1.13±0.13^b	2.19±0.24^b
M-Med组	0.91±0.10^bc	1.73±0.21^bc
H-Med组	0.68±0.06^bcd	1.21±0.13^bcd
H-Med+DMOG组	1.15±0.12^e	2.25±0.22^e
F	39.689^＊＊	52.596^＊＊