Protective functions of Lycium barbarum polysaccharides in H2O2‑injured vascular endothelial cells through anti‑oxidation and anti‑apoptosis effects

Xue,Shujing; Hu,Xiaohui; Zhu,Lingin; Nie,Lihong; Li,Guanghua

doi:10.3892/br.2019.1240

Protective functions of Lycium barbarum polysaccharides in H2O2‑injured vascular endothelial cells through anti‑oxidation and anti‑apoptosis effects

Authors:
- Shujing Xue
- Xiaohui Hu
- Lingin Zhu
- Lihong Nie
- Guanghua Li
View Affiliations

Affiliations: School of Basic Medical Science, Ningxia Medical University, Yinchuan, Ningxia 750004, P.R. China, School of Public Health and Management, Ningxia Medical University, Yinchuan, Ningxia 750004, P.R. China
Published online on: September 10, 2019 https://doi.org/10.3892/br.2019.1240
Pages: 207-214
Copyright: © Xue et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Cell injury in the cardiovascular endothelia caused by oxidative stress is among the major inducers of endothelium dysfunction and serves an important role in initiating cardiovascular diseases (CVDs). Therefore, protecting and improving the normal function of endothelial cells are considered key measures against CVDs. As a traditional Chinese medicinal component, Lycium barbarum is regarded to have high medicinal value. The present study aimed to investigate the potential anti‑apoptosis and anti‑oxidation effects of Lycium barbarum polysaccharides (LBPs) on injured rat artery endothelial cells, to demonstrate the experimental and medicinal values of LBPs. In the present study, the aortic endothelial cells of rats were cultivated and randomly divided into five groups: A control group, H2O2‑injured group (H2O2 group), H2O2+LBPs (110 µg/ml) group (low‑dose group, LT), H2O2+LBPs (220 µg/ml) group (medium‑dose group, MT) and H2O2+LBPs (440 µg/ml) group (high‑dose group, HT). Among these, the activity of superoxide dismutase (SOD), and the levels of malondialdehyde (MDA) and nitric oxide (NO) were detected by colorimetry. Additionally, the expression of B‑cell lymphoma‑2 (Bcl‑2) and Bcl‑2‑associated X protein (Bax) were detected by western blotting. It was observed that SOD activity and NO content decreased while MDA content increased significantly in the H2O2 group (P<0.05 vs. control); that SOD activity in the MT and HT group, and NO content in all three LBP groups were increased, while MDA content in the three LBP groups was decreased, compared with the H2O2 group (all P<0.05); that Bcl‑2 expression decreased significantly in the H2O2 group while the expression of Bax increased significantly compared with the control group (both P<0.05); and that Bcl‑2 expression in all three LBP groups increased, while Bax expression in the MT and HT groups decreased compared with the H2O2 group (all P<0.05), with these altered Bax levels being statistically similar to those in the control group (P>0.05). On light microscopy, the cells in the control group exhibited spindle‑shaped morphology, consistent sizes, defined boundaries, and distinct nuclei of equivalent sizes with round or oval morphology. Additionally, the chromatin in the nuclei was evenly distributed, and all cells were adhered in a paving‑stone arrangement. Notably, only few cells died. Conversely, the cells in the H2O2 group exhibited signs of damage and enlarged gaps, and focal cells died. In the HT group, the cells once again appeared adherent and exhibited similar morphological status to the normal cells. Overall, these results indicate that LBPs serve a protective role in oxidative‑injured vascular endothelial cells through anti‑apoptosis and anti‑oxidation effects.

View Figures

View References

1	Mladěnka P, Applová L, Patočka J, Costa VM, Remiao F, Pourová J, Mladěnka A, Karlíčková J, Jahodář L, Vopršalová M, et al: Comprehensive review of cardiovascular toxicity of drugs and related agents. Med Res Rev. 38:1332–1403. 2018.PubMed/NCBI View Article : Google Scholar
2	Kim KS, Song CG and Kang PM: Targeting oxidative stress using nanoparticles as atheranostic strategy for cardiovascular diseases. Antioxid Redox Signal. 30:733–746. 2019.PubMed/NCBI View Article : Google Scholar
3	World Health Organization: Comprehensive global monitoring framework, including indicators, and a set of voluntary global targets for the prevention and control of noncommunicable diseases. World Health Organization, 2012.
4	Zeng Z: Internal secretion of vascular endothelial cells. Chin J Int Med. 37:77. 1998.(In Chinese)http://www.cqvip.com/Main/Detail.aspx?id=2964468.
5	Blatter LA: Tissue specificity: SOCE: Implications for Ca²⁺ handling in endothelial cells. Adv Exp Med Biol. 993:343–361. 2017.PubMed/NCBI View Article : Google Scholar
6	Rajendran P, Rengarajan T, Thangavel J, Nishigaki Y, Sakthisekaran D, Sethi G and Nishigaki I: The vascular endothelium and human diseases. Int J Biol Sci. 9:1057–1069. 2013.PubMed/NCBI View Article : Google Scholar
7	Yamada T, Egashira N, Bando A, Nishime Y, Tonogai Y, Imuta M, Yano T and Oishi R: Activation of p38 MAPK by oxidative stress underlying epirubicin-induced vascular endothelial cell injury. Free Radic Boil Med. 52:1285–1293. 2012.PubMed/NCBI View Article : Google Scholar
8	Closhen D, Bender B, Luhmann HJ and Kuhlmann CR: CRP-induced levels of oxidative stress are higher in brain than aortic endothelial cells. Cytokine. 50:117–120. 2010.PubMed/NCBI View Article : Google Scholar
9	Higashi Y: Mechanisms of impairment of endothelial cell. Nihon Rinsho. 70:1519–1523. 2012.(In Japanese). PubMed/NCBI
10	Bei Y, Das S, Rodosthenous RS, Holvoet P, Vanhaverbeke M, Monteiro MC, Monteiro VVS, Radosinska J, Bartekova M, Jansen F, et al: Extracellular vesicles in cardiovascular theranostics. Theranostics. 7:4168–4182. 2017.PubMed/NCBI View Article : Google Scholar
11	Rodrigo R, Libuy M, Feliú F and Hasson D: Oxidative stress-related biomarkers in essential hypertension and ischemia-reperfusion myocardial damage. Dis Markers. 35:773–790. 2013.PubMed/NCBI View Article : Google Scholar
12	Xu J and Li Y: Effects of salidroside on exhaustive exercise-induced oxidative stress in rats. Mol Med Rep. 6:1195–1198. 2012.PubMed/NCBI View Article : Google Scholar
13	Luo Q, Li Z, Huang X, Yan J, Zhang S and Cai YZ: Lycium barbarum polysaccharides: Protective effects against heat-induced damage of rat testes and H₂O₂-induced DNA damage in mouse testicular cells and beneficial effect on sexual behavior and reproductive function of hemicastrated rats. Life Sci. 79:613–621. 2006.PubMed/NCBI View Article : Google Scholar
14	Ho YS, Yu MS, Yik SY, So KF, Yuen WH and Chang RC: Polysaccharides from wolfberry antagonizes glutamate excitotoxicity in rat cortical neurons. Cell Mol Neurobiol. 29:1233–1244. 2009.PubMed/NCBI View Article : Google Scholar
15	Wu HT, He XJ, Hong YK, Ma T, Xu YP and Li HH: Chemical characterization of Lycium barbarum polysaccharides and its inhibition against liver oxidative injury of high-fat mice. Int J Biol Macromol. 46:540–543. 2010.PubMed/NCBI View Article : Google Scholar
16	Tang WM, Chan E, Kwok CY, Lee YK, Wu JH, Wan CW, Chan RY, Yu PH and Chan SW: A review of the anticancer and immunomodulatory effects of Lycium barbarum fruit. Inflammopharmacology. 20:307–314. 2012.PubMed/NCBI View Article : Google Scholar
17	Chang HM But PPH (eds): Pharmacology and Applications of Chinese Materia Medica, Vol. II. World Scientific Publishing Company Incorporated, Singapore, 1986. https://www.worldscientific.com/worldscibooks/10.1142/0377.
18	Potterat O: Goji (Lycium barbarum and L. chinense): Phytochemistry, pharmacology and safety in the perspective of traditional uses and recent popularity. Planta Med. 76:7–19. 2010.PubMed/NCBI View Article : Google Scholar
19	Yu N, Song N, Liu CY and Yang GL: The estrogen-like protective effect of Lycium barbarum polysaccharides in reducing oxidative stress on myocardial cells from ovariectomized rats. Molecular Medicine Reports. 19:2271–2278. 2019.PubMed/NCBI View Article : Google Scholar
20	Li J, Zou Y, Ge J, Zhang D, Guan A, Wu J and Li L: The effects of G-CSF on proliferation of mouse myocardial microvascular endothelial cells. Int J Mol Sci. 12:1306–1315. 2011.PubMed/NCBI View Article : Google Scholar
21	Younus A, Aneni EC, Spatz ES, Osondu CU, Roberson L, Ogunmoroti O, Malik R, Ali SS, Aziz M, Feldman T, et al: A systematic review of the prevalence and outcomes of ideal cardiovascular health in US and Non-US populations. Mayo Clin Proc. 91:649–670. 2016.PubMed/NCBI View Article : Google Scholar
22	Fang N, Jiang M and Fan Y: Ideal cardiovascular health metrics and risk of cardiovascular disease or mortality: A meta-analysis. Int J Cardiol. 214:279–283. 2016.PubMed/NCBI View Article : Google Scholar
23	Guo L and Zhang S: Association between ideal cardiovascular health metrics and risk of cardiovascular events or mortality: A meta-analysis of prospective studies. Clin Cardiol. 40:1339–1346. 2017.PubMed/NCBI View Article : Google Scholar
24	Yang D, Li SY, Yeung CM, Chang RC, So KF, Wong D and Lo AC: Lycium barbarum extracts protect the brain from blood-brain barrier disruption and cerebral edema in experimental stroke. PLoS One. 7(e33596)2012.PubMed/NCBI View Article : Google Scholar
25	Shan X, Zhou J, Ma T and Chai Q: Lycium barbarum polysaccharides reduce exercise-induced oxidative stress. Int J Mol Sci. 12:1081–1088. 2011.PubMed/NCBI View Article : Google Scholar
26	Teng P, Li Y, Cheng W, Zhou L, Shen Y and Wang Y: Neuro-protective effects of lycium barbarum polysaccharides in lipopolysaccharide-induced BV2 microglial cells. Mol Med Rep. 7:1977–1981. 2013.PubMed/NCBI View Article : Google Scholar
27	Chang RC and So KF: Use of anti-aging herbal medicine, Lycium barbarum, against aging-associated diseases. What do we know so far? Cell Mol Neurobiol. 28:643–652. 2008.PubMed/NCBI View Article : Google Scholar
28	Liu Y and Zhang Y: Lycium barbarum polysaccharides alleviate hydrogen peroxide-induced injury by up-regulation of miR-4295 in human trabecular meshwork cells. Exp Mol Pathol. 106:109–115. 2019.PubMed/NCBI View Article : Google Scholar
29	Niu T, Jin L, Niu S, Gong C and Wang H: Lycium barbarum polysaccharides alleviates oxidative damage induced by H₂O₂ through down-regulating MicroRNA-194 in PC-12 and SH-SY5Y cells. Cell Physiol Biochem. 50:460–472. 2018.PubMed/NCBI View Article : Google Scholar
30	Qi B, Ji Q, Wen Y, Liu L, Guo X, Hou G, Wang G and Zhong J: Lycium barbarum polysaccharides protect human lens epithelial cells against oxidative stress-induced apoptosis and senescence. PLoS One. 9(e110275)2014.PubMed/NCBI View Article : Google Scholar
31	Schulz C, Farkas L, Wolf K, Kratzel K, Eissner G and Pfeifer M: Differences in LPS-induced activation of bronchial epithelial cells (BEAS-2B) and type II-like pneumocytes (A-549). Scand J Immunol. 56:294–302. 2002.PubMed/NCBI View Article : Google Scholar
32	Zhao W, Pan X, Li T, Zhang C and Shi N: Lycium barbarum polysaccharides protect against trimethyltin chloride-induced apoptosis via sonic hedgehog and PI3K/Akt signaling pathways in mouse neuro-2a cells. Oxid Med Cell Longev. 2016(9826726)2016.PubMed/NCBI View Article : Google Scholar
33	Zhang Y, Wang G, Wang T, Cao W, Zhang L and Chen X: Nrf2-Keap1 pathway-mediated effects of resveratrol on oxidative stress and apoptosis in hydrogen peroxide-treated rheumatoid arthritis fibroblast-like synoviocytes. Ann N Y Acad Sci. 6:2019.https://doi.org/10.1111/nyas.14196. PubMed/NCBI View Article : Google Scholar
34	Maritim AC, Sanders RA and Watkins JB III: Diabetes, oxidative stress and antioxidants: A review. J Biochem Mol Toxicol. 17:24–38. 2003.PubMed/NCBI View Article : Google Scholar
35	Wang Z, Su G, Zhang Z, Dong H, Wang Y, Zhao H, Zhao Y and Sun Q: 25-Hydroxyl-protopanaxatriol protects against H₂O₂-induced H9c2 cardiomyocytes injury via PI3K/Akt pathway and apoptotic protein down-regulation. Biomed Pharmacother. 99:33–42. 2018.PubMed/NCBI View Article : Google Scholar
36	Zhao Z, Luo Y, Li G, Zhu L, Wang Y and Zhang X: Thoracic aorta vasoreactivity in rats under exhaustive exercise: Effects of Lycium barbarum polysaccharides supplementation. J Int Soc Sports Nutr. 10(47)2013.PubMed/NCBI View Article : Google Scholar
37	Yang CH, Xu JH, Ren QC, Duan T, Mo F and Zhang W: Melatonin promotes secondary hair follicle development of early post-natal cashmere goat and improves cashmere quantity and quality by enhancing antioxidant capacity and suppressing apoptosis. J Pineal Res. 67(e12569)2019.PubMed/NCBI View Article : Google Scholar
38	Majumdar A and Kar RK: Orchestration of Cu-Zn SOD and class III peroxidase with upstream interplay between NADPH oxidase and PM H⁺-ATPase mediates root growth in Vigna radiata (L.) Wilczek. J Plant Physiol. 232:248–256. 2019.PubMed/NCBI View Article : Google Scholar
39	Liochev SI: Reactive oxygen species and the free radical theory of aging. Free Radic Biol Med. 60:1–4. 2013.PubMed/NCBI View Article : Google Scholar
40	Kumar M, Chand R and Shah K: Evidences for growth-promoting and fungicidal effects of low doses of tricyclazole in barley. Plant Physiol Biochem. 103:176–182. 2016.PubMed/NCBI View Article : Google Scholar
41	Lu Z, Xu X, Hu X, Zhu G, Zhang P, van Deel ED, French JP, Fassett JT, Oury TD, Bache RJ and Chen Y: Extracellular superoxide dismutase deficiency exacerbates pressure overload-induced left ventricular hypertrophy and dysfunction. Hypertension. 51:19–25. 2008.PubMed/NCBI View Article : Google Scholar
42	Song QQ, Niu JP, Zhang SY, Liang TT, Zhou J and Feng SS: Effects of simulated heat wave and ozone on high fat diet ApoE deficient mice. Biomed Environ Sci. 31:757–768. 2018.PubMed/NCBI View Article : Google Scholar
43	Shatoor AS, Al Humayed S, Alkhateeb MA, Shatoor KA, Aldera H, Alassiri M and Shati AA: Crataegus Aronia protects and reverses vascular inflammation in a high fat diet rat model by an antioxidant mechanism and modulating serum levels of oxidized low-density lipoprotein. Pharm Biol. 57:38–48. 2019.PubMed/NCBI View Article : Google Scholar
44	Saran M, Michel C and Bors W: Radical functions in vivo: A critical review of current concepts and hypotheses. Z Naturforsch C. 53:210–227. 1998.PubMed/NCBI View Article : Google Scholar
45	Bai YX, Fang F, Jiang JL and Xu F: Extrinsic calcitonin gene-related peptide inhibits hyperoxia-induced alveolar epithelial type II cells apoptosis, oxidative stress, and reactive oxygen species (ROS) production by enhancing notch 1 and homocysteine-induced endoplasmic reticulum protein (HERP) expression. Med Sci Monit. 23:5774–5782. 2017.PubMed/NCBI View Article : Google Scholar
46	Omer FAA, Hashim NBM, Ibrahim MY, Dehghan F, Yahayu M, Karimian H, Salim LZA and Mohan S: Beta-mangostin from Cratoxylum arborescens activates the intrinsic apoptosis pathway through reactive oxygen species with downregulation of the HSP70 gene in the HL60 cells associated with a G₀/G₁ cell-cycle arrest. Tumour Biol. 39(1010428317731451)2017.PubMed/NCBI View Article : Google Scholar
47	Radović N, Cucić S and Altarac S: Molecular aspects of apoptosis. Acta Med Croatica. 62:249–256. 2008.(In Croatian). PubMed/NCBI
48	Qi B, Ji Q, Wen Y, Liu L, Guo X, Hou G, Wang G and Zhong J: Lycium barbarum polysaccharides protect human lens epithelial cells against oxidative stress-induced apoptosis and senescence. PLoS One. 9(10)(e110275)2014.PubMed/NCBI View Article : Google Scholar
49	Edlich F: BCL-2 proteins and apoptosis: Recent insights and unknowns. Biochem Biophys Res Commun. 500:26–34. 2018.PubMed/NCBI View Article : Google Scholar
50	Xi H, Fan X, Zhang Z, Liang Y, Li Q and He J: Bax and Bcl-2 are involved in the apoptosis induced by local testicular heating in the boar testis. Reprod Domest Anim. 52:359–365. 2017.PubMed/NCBI View Article : Google Scholar
51	Gross A: BCL-2 family proteins as regulators of mitochondria metabolism. Biochim Biophys Acta. 1857:1243–1246. 2016.PubMed/NCBI View Article : Google Scholar
52	Wang X, Li Z, Bai J, Song W and Zhang F: miR-17-5p regulates the proliferation and apoptosis of human trabecular meshwork cells by targeting phosphatase and tensin homolog. Mol Med Rep. 19:3132–3138. 2019.PubMed/NCBI View Article : Google Scholar
53	Goping IS, Gross A, Lavoie JN, Nguyen M, Jemmerson R, Roth K, Korsmeyer SJ and Shore GC: Regulated targeting of BAX to mitochondria. J Cell Biol. 143:207–215. 1998.PubMed/NCBI View Article : Google Scholar
54	Cheng EH, Wei MC, Weiler S, Flavell RA, Mak TW, Lindsten T and Korsmeyer SJ: Bcl-2, Bcl-X_L sequester BH3 domain-only molecules preventing Bax-and Bak-mediated mitochondrial apoptosis. Mol Cell. 8:705–711. 2001.PubMed/NCBI View Article : Google Scholar
55	Yang J, Liu X, Bhalla K, Kim CN, Ibrado AM, Cai J, Peng TI, Jones DP and Wang X: Prevention of apoptosis by Bcl-2: Release of cytochrome C from mitochondria blocked. Science. 275:1129–1132. 1997.PubMed/NCBI View Article : Google Scholar
56	Hsu HC, Liu YS, Tseng KC, Tan BC Chen SJ and Chen HC: Lgr5 regulates survival through mitochondria-mediated apoptosis and by targeting the Wnt/β-catenin signaling pathway in colorectal cancer cells. Cellular Signalling. 26:2333–2342. 2014.PubMed/NCBI View Article : Google Scholar