黄芪多糖对野百合碱诱导的肺动脉高压大鼠肺动脉血管的保护作用及其机制

doi:10.13481/j.1671-587X.20210605

Abstract

Abstract: Objective

To investigate the intervention effect of astragalus polysaccharide （APS） on the pulmonary vascular inflammation of the rats with pulmonary hypertension （PAH） induced by monocrotaline （MCT）， and to clarify its mechanism.

Methods

Sixty male SD rats were randomly divided into control group， PAH group， Calpain-1 inhibitor group， NOD like receptor family protein 3 （NLRP3） inhibitor group， low dose of APS group（400 mg·kg^－1APS），and high dose of APS group（800 mg·kg^－1 APS）.The PAH rat model was established by intraperitoneal injection of 60 mg·kg^－1MCT.The serum levels of interleukin-18 （IL-18） and interleukin-1 β（IL-1 β）of the rats in various groups were detected by ELISA method. HE staining was used to observe the pathomorphology of lung tissue of the rats in various groups.The right ventricular systolic pressure （RVSP）， right ventricular hypertrophy index （RVHI），the percentage of pulmonary arteriole wall thickness in total vessel diameter （WT%） and the percentage of pulmonary arteriole wall area in total vessel area （WA%）of the rats in various groups were measured.Immunohistochemistry staining was used to detect the expressions of Calpain-1 in lung tissue of the rats in various groups.The expression levels of B-cell lymphoma-2（Bcl-2），Bcl-2 assaciated X protein（Bax）， Caspase-3， NLRP3， apoptosis related particulate protein （ASC），Caspase-1， and Calpain-1 in lung tissue of the rats in various groups were detected by Western blotting method.

Results

Compared with PAH group， the serum levels of IL-18 and IL-1β in low and high doses of APS groups and Calpain-1 inhibitor group were decreased （P<0.05）. The HE staining results showed that the morphology of lung tissue of the rats in control group was normal， and the cells did not fall off and proliferate， and there was no inflammatory cell infiltration； in PAH group， the endothelium of pulmonary arterioles fell off， the smooth muscle cells proliferated seriously， and a large number of inflammatory cells infiltrated in the alveolar cavity；the endothelium of pulmonary arterioles in low and high doses of APS groups， Calpain-1 inhibitor group and NLRP3 inhibitor group fell off slightly， the proliferation of smooth muscle cells and inflammatory cell infiltration were decreased. Compared with PAH group， the RVSP and RVHI of the rats in low and high doses of APS groups， Calpain-1 inhibitor group and NLRP3 inhibitor group were decreased significantly （P<0.05）.The immunohistochemistry staining results showed that compared with PAH group， the expression amounts of Calpain-1 in lung tissue of the rats in low and high doses of APS groups and Calpain-1 inhibitor group were decreased.Compared with PAH group，the Calpain-1 activities in lung tissue of the rats in low and high doses of APS groups and Calpain-1 inhibitor group were decreased（P<0.05）. The Western blotting results showed that compared with PAH group， the expression levels of Bax，Caspase-3，NLRP3， ASC，Caspase-1，and Calpain-1 proteins in lung tissue of the rats in low and high doses of APS groups， Calpain-1 inhibitor group， and NLRP3 inhibitor group were decreased （P<0.05）， and the expression levels of Bcl-2 protein were increased （P<0.05）.

Conclusion

APS has a protective effect on the pulmonary arteries of the PAH rats induced by MCT， and its mechanism may be related to the effect of APS on the expression of Calpain-1 protein in lung tissue of the PAH rats.

Key words: astragalus polysaccharides, pulmonary arteries hypertension, monocrotaline, Calpain-1, NOD like receptor family protein 3, inflammasome

CLC Number:

R543.2

Cong LI,Kun ZHAO,Jingliang ZHANG,Yingjie ZHANG,Hongxin WNAG. Protective effect of astragalus polysaccharides on pulmonary artery vessels in rats with MCT-induced pulmonary hypertension and its mechanism[J].Journal of Jilin University(Medicine Edition), 2021, 47(6): 1371-1379.

Figures/Tables 12

Tab.1

Fig. 1

Tab.2

Fig. 2

Tab.3

Fig.3

Tab.4

Tab.5

Fig. 4

Fig.5

Tab.6

Fig. 6

References 22

1	LEOPOLD J， MARON B. Molecular mechanisms of pulmonary vascular remodeling in pulmonary arterial hypertension［J］. Int J Mol Sci， 2016， 17（5）： 761.
2	KIM D， GEORGE M P. Pulmonary hypertension［J］. Med Clin N Am， 2019， 103（3）： 413-423.
3	MATHEW R. Pathogenesis of pulmonary hypertension： a case for caveolin-1 and cell membrane integrity［J］. Am J Physiol Heart Circ Physiol， 2014， 306（1）： H15-H25.
4	KELLEY N， JELTEMA D， DUAN Y H， et al. The NLRP3 inflammasome： an overview of mechanisms of activation and regulation［J］.Int J Mol Sci，2019，20（13）： 3328.
5	KANG Y， ZHANG G Y， HUANG E C， et al. Sulforaphane prevents right ventricular injury and reduces pulmonary vascular remodeling in pulmonary arterial hypertension［J］. Am J Physiol Heart Circ Physiol， 2020， 318（4）： H853-H866.
6	PASQUA T， PAGLIARO P， ROCCA C， et al. Role of NLRP-3 inflammasome in hypertension： a potential therapeutic target［J］. Curr Pharm Biotechnol， 2018， 19（9）： 708-714.
7	CHEN J X， WU Y Z， ZHANG L M， et al. Evidence for Calpains in cancer metastasis［J］. J Cell Physiol， 2019， 234（6）： 8233-8240.
8	YU L， YIN M H， YANG X Y， et al. Calpain inhibitor Ⅰ attenuates atherosclerosis and inflammation in atherosclerotic rats through eNOS/NO/NF-κB pathway［J］. Can J Physiol Pharmacol， 2018， 96（1）： 60-67.
9	YU Y， SHI H， YU Y， et al. Inhibition of Calpain alleviates coxsackievirus B3-induced myocarditis through suppressing the canonical NLRP3 inflammasome/caspase-1-mediated and noncanonical caspase-11-mediated pyroptosis pathways［J］. Am J Transl Res， 2020， 12（5）： 1954-1964.
10	YUAN L B， HUA C Y， GAO S， et al. Astragalus polysaccharides attenuate monocrotaline-induced pulmonary arterial hypertension in rats［J］. Am J Chin Med， 2017， 45（4）： 773-789.
11	LI W， HU X， WANG S， et al. Characterization and anti-tumor bioactivity of astragalus polysaccharides by immunomodulation［J］. Int J Biol Macromol， 2020， 145： 985-997.
12	VÄLIMÄKI E， CYPRYK W， VIRKANEN J， et al. Calpain activity is essential for ATP-driven unconventional vesicle-mediated protein secretion and inflammasome activation in human macrophages［J］. J Immunol， 2016， 197（8）： 3315-3325.
13	TAPIA V S， DANIELS M J D， PALAZÓN-RIQUELME P， et al. The three cytokines IL-1β， IL-18， and IL-1α share related but distinct secretory routes［J］. J Biol Chem， 2019， 294（21）： 8325-8335.
14	DORFMÜLLER P， HUMBERT M， PERROS F，et al. Fibrous remodeling of the pulmonary venous system in pulmonary arterial hypertension associated with connective tissue diseases［J］.Hum Pathol，2007，38（6）：893-902.
15	XIAO R， ZHU L P， SU Y， et al. Monocrotaline pyrrole induces pulmonary endothelial damage through binding to and release from erythrocytes in lung during venous blood reoxygenation［J］. Am J Physiol Lung Cell Mol Physiol， 2019， 316（5）： L798-L809.
16	ZHANG D， WANG X L， CHEN S Y， et al. Endogenous hydrogen sulfide sulfhydrates IKKβ at cysteine 179 to control pulmonary artery endothelial cell inflammation［J］. Clin Sci， 2019， 133（20）： 2045-2059.
17	TANG B， CHEN G X， LIANG M Y， et al. Ellagic acid prevents monocrotaline-induced pulmonary artery hypertension via inhibiting NLRP3 inflammasome activation in rats［J］. Int J Cardiol， 2015， 180： 134-141.
18	ZHANG M， XIN W， YU Y， et al. Programmed death-ligand 1 triggers PASMCs pyroptosis and pulmonary vascular fibrosis in pulmonary hypertension［J］. J Mol Cell Cardiol， 2020， 138： 23-33.
19	CAO T， FAN S， ZHENG D， et al. Increased Calpain-1 in mitochondria induces dilated heart failure in mice： role of mitochondrial superoxide anion［J］. Basic Res Cardiol， 2019， 114（3）： 1-15.
20	CHEN K， HE L， LI Y， et al. Inhibition of GPR35 preserves mitochondrial function after myocardial infarction by targeting Calpain 1/2［J］. J Cardiovasc Pharmacol， 2020， 75（6）： 556-563.
21	YUE R C， LU S Z， LUO Y， et al. Calpain silencing alleviates myocardial ischemia-reperfusion injury through the NLRP3/ASC/Caspase-1 axis in mice［J］. Life Sci， 2019， 233： 116631.
22	SUN Y， LU M， SUN T， et al. Astragaloside Ⅳ attenuates inflammatory response mediated by NLRP-3/Calpain-1 is involved in the development of pulmonary hypertension［J］. J Cell Mol Med， 2021， 25（1）： 586-590.

Group	n	RVSP（P/mmHg）	RVHI
Control	6	13.1±0.55	0.21±0.02
PAH	7	28.9±1.35^*	0.53±0.03^*
Calpain-1 inhibitor	7	20.2±0.37^△	0.38±0.01^△
NLRP3 inhibitor	7	18.3±1.02^△	0.31±0.02^△
APS Low dose	7	21.2±0.15^△	0.41±0.01^△
High dose	8	17.9±0.75^△	0.32±0.02^△

Group	WT%	WA%
Control	18.32±1.56	22.45±1.67
PAH	55.15±2.23^*	60.45±3.32^*
Calpain-1 inhibitor	36.34±2.41^△	41.34±2.34^△
NLRP3 inhibitor	25.45±2.65^△	28.46±2.56^△
APS Low dose	34.98±2.12^△	42.78±2.78^△
High dose	26.96±2.24^△	33.12±1.23^△

Group	Bcl-2	Bax	Caspase-3
Control	0.78±0.05	0.38±0.04	0.31±0.06
PAH	0.37±0.03^*	0.84±0.05^*	0.88±0.08^*
Calpain-1 inhibitor	0.57±0.04^△	0.67±0.04^△	0.53±0.05^△
NLRP3inhibitor	0.62±0.03^△	0.58±0.03^△	0.44±0.04^△
APS Low dose	0.63±0.03^△	0.62±0.05^△	0.58±0.03^△
High dose	0.71±0.05^△	0.54±0.04^△	0.49±0.03^△

Group	NLRP3	ASC	Caspase-1
Control	0.39±0.05	0.31±0.04	0.39±0.04
PAH	0.82±0.06^*	0.87±0.04^*	0.92±0.06^*
Calpain-1 inhibitor	0.59±0.06^△	0.54±0.04^△	0.62±0.06^△
NLRP3 inhibitor	0.48±0.04^△	0.43±0.02^△	0.48±0.03^△
APS Low dose	0.58±0.05^△	0.68±0.05^△	0.63±0.03^△
High dose	0.53±0.02^△	0.61±0.02^△	0.44±0.04^△

Group	IL-1β	IL-18
Control	48.04±2.53	16.89±2.13
PAH	138.34±5.32^*	96.39±3.78^*
Calpain-1 inhibitor	95.89±6.67^△	69.23±3.19^△
NLRP3 inhibitor	72.47±5.65^△	55.81±3.71^△
APS Low dose	99.82±5.34^△	68.49±3.32^△
High dose	75.28±4.56^△	52.78±2.12^△

Protective effect of astragalus polysaccharides on pulmonary artery vessels in rats with MCT-induced pulmonary hypertension and its mechanism

RichHTML

PDF (PC)

Like

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 12

References 22

Related Articles 5

Metrics

Comments

Recommended 10

Group	Calpain-1 protein
Control	0.29±0.08
PAH	0.74±0.04^*
NLRP3 inhibitor	0.69±0.06
Calpain-1 inhibitor	0.42±0.11^△
APS
Low dose	0.48±0.07^△
High dose	0.38±0.05^△

[1]	WANG Lihong, ZHANG Ying, LAN Kun, ZHANG Haiyu, CAO Xiaobei, LI Shanyu. Inhibitory effect of astragalus polysaccharides on pulmonary inflammation in asthma model mice and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2019, 45(02): 313-318.
[2]	HU Bo, WANG Xiaowen, CAO Jianhui, SUN Xiaomin, CUI Yajie, SHI Congcong. Changes of expressions of NLRP3 inflammasome in peripheral blood mononuclear cells and IL-1β and IL-18 in serum in children with asthma and their significances [J]. Journal of Jilin University(Medicine Edition), 2019, 45(01): 111-116.
[3]	SONG Chenxue, WANG Yubo, LIU Chuangui, XIE Jingshu, LU Yanjiao, WANG Ting, WANG Guoqiang, WANG Yawei, WANG Fang, ZHENG Jingtong. Treatment of Helicobacter pylori induced chronic atrophic gastritiswith traditional Chinese medicine combined standard triple therapyand its mechanisms [J]. Journal of Jilin University Medicine Edition, 2016, 42(04): 789-792.
[4]	ZHAO Ke-Xing, SUN Jiang-Bin, LIU Ke-Xiang, WU Hui-Ying, LI Bo. Comparison of pathological changes of lung tissue in rat pulmonary arterial hypertension model induced by two different doses of monocrotaline [J]. J4, 2009, 35(5): 870-873.
[5]	ZHANG Hai-Yu, PIAO Hui-Shun, JIN Zai-Jiu, ZHANG Shan-Yu. Effects of mixture from ginsebosides and astragalus polysaccharides on immune and antioxidative function in immunosuppressive mice induced with CTX [J]. J4, 2009, 35(5): 857-861.