茯苓多糖对LPS诱导猫肾细胞炎症的保护作用研究

doi:10.16431/j.cnki.1671-7236.2023.11.041

Abstract

Abstract: 【Objective】 This study was aimed to investigate the protective effect of Poria cocos polysaccharide (PCP)on lipopolysaccharide (LPS) induced inflammation in crandell rees feline kidney (CRFK) and its anti-inflammatory mechanism.【Method】 MTT method was used to detect the changes of cell activity by different concentrations of PCP (5, 10, 15, 25, 35 and 45 μg/mL) and LPS (20, 40, 60, 80, 100, 125 and 150 μg/mL) on CRFK.The LPS-induced CRFK inflammation model was treated with different concentrations of PCP to observe the morphological changes of cells.Nitric oxide (NO) release and cell apoptosis in CRFK were detected by nitric reductase method and flow cytometry, respectively.Real-time quantitative PCR and Western blotting were used to detect the expression levels of genes and proteins related to cell inflammation and apoptosis.【Result】 5-25 μg/mL PCP was non-toxic to CRFK and could reverse the cell morphological changes of CRFK after LPS stimulation.25 μg/mL PCP significantly decreased NO release and apoptosis rate in CRFK after LPS stimulation (P<0.05).Real-time quantitative PCR results showed that different concentrations of PCP could significantly reduce the mRNA expressions of IL-6, TNF-α, Caspase3, Caspase8, Caspase9, Bid and Bax in LPS-induced CRFK, and significantly up-regulate the mRNA expression of Bcl-2 (P<0.05).Western blotting results showed that the expression levels of TLR4, NF-κB, Caspase3 and Caspase9 proteins in LPS-induced CRFK were significantly increased (P<0.05), but after PCP treatment, the expression levels of TLR4, NF-κB, Caspase3 and Caspase9 proteins were significantly decreased (P<0.05).【Conclusion】 PCP could inhibit LPS-induced cell inflammatory damage of CRFK by regulating apoptosis, and its mechanism might be related to the inhibition of TLR4-NF-κB signaling pathway.The results provided a new target and experimental basis for the treatment of CRFK inflammation by PCP.

Key words: Poria cocos polysaccharide; crandell reese feline kidney; lipopolysaccharide; inflammatory; apoptosis

CLC Number:

S859.7

LIU Jiali, LIN Bing, LIU Xin, YAN Pupu, XIA Jinjin, HUANG Yongxi, BAI Ruonan, GUO Liwei, LIU Guoping. Protective Effect of Poria cocos Polysaccharides on LPS-induced CRFK Cell Inflammation[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(11): 4747-4758.

References

[1] 王路瑶, 胡继划, 唐焓嫣, 等.中药抑菌作用特点及其开发优势研究[J].国外医药(抗生素分册), 2023, 44(2):91-95. WANG L Y, HU J H, TANG H Y, et al.Study on the characteristics of antibacterial effect of traditional Chinese medicine and its development advantages[J] World Notes on Antibiotics, 2023, 44(2):91-95.(in Chinese)
[2] SHIMODE S, SAKUMA T, YAMAMOTO T, et al.Establishment of CRFK cells for vaccine production by inactivating endogenous Retrovirus with TALEN technology[J].Scientific Reports, 2022, 12(1):6641.
[3] QIU J, CHENG F, BURGER L R, et al.The transcription profile of Aleutian mink disease virus in CRFK cells is generated by alternative processing of pre-mRNAs produced from a single promoter[J].Journal of Virology, 2006, 80(2):654-662.
[4] LIU Y, SHI W D, XIE Q Q, et al.Induction of COX-2 by Feline calicivirus via activation of the MEK1-ERK1/2 pathway, and attenuation of feline lung inflammation and injury by MEK1 inhibitor AZD6244(selumetinib)[J].Biochemical and Biophysical Research Communications, 2022, 604:8-13.
[5] DRECHSLER Y, VASCONCELOS E J R, GRIGGS L M, et al.CRFK and primary macrophages transcriptomes in response to Feline coronavirus infection differ significantly[J].Frontiers in Genetics, 2020, 11:584744.
[6] 王宝刚, 杨立军, 曹鹏亮, 等.不同处理方法对中草药体外抑菌及抗氧化活性的影响[J].中国饲料, 2023, 719(3):71-76. WANG B G, YANG L J, CAO P L, et al.Effects of treatment methods on bacteriostatic and antioxidant activities of Chinese herbal medicine in vitro[J] China Feed, 2023, 719(3):71-76.(in Chinese)
[7] 李瑞国, 杨洁, 石柏灌, 等.咀嚼食品中草药添加对宠物犬血液生化指标及肠道菌群的影响[J].中国饲料, 2022, 697(5):100-106. LI R G, YANG J, SHI B G, et al.Effects of Chinese herbal medicine on blood biochemical indexes and intestinal microflora in pet dogs[J] China Feed, 2022, 697(5):100-106.(in Chinese)
[8] 孙秀红.中草药及中草药饲料添加剂的应用前景分析[J].畜禽业, 2015, 3:24-26. SUN X H.Application prospect analysis of Chinese herbal medicines and Chinese herbal feed additives[J] Livestock and Poultry Industry, 2015, 3:24-26.(in Chinese)
[9] 陈成.食源性真菌多糖在功能性食品中的应用[J].江苏调味副食品, 2016, 145(2):6-8. CHEN C.The application of food-borne fungus polysaccharose in functional food[J] Jiangsu Condiment and Subsidiary Food, 2016, 145(2):6-8.(in Chinese)
[10] 赵书远, 高雪琴.食源性真菌多糖在功能性食品中的应用[J].食品安全导刊, 2020, 276(17):45. ZHAO S Y, GAO X Q.The application of food-borne fungus polysaccharose in functional food[J] China Food Safety Magazine, 2020, 276(17):45.(in Chinese)
[11] KWON E B, OH Y C, HWANG Y H, et al.A herbal mixture formula of OCD20015-V009 prophylactic administration to enhance interferon-mediated antiviral activity against Influenza A virus[J].Frontiers in Pharmacology, 2021, 12:764297.
[12] YI Y, HUA H, SUN X, et al.Rapid determination of polysaccharides and antioxidant activity of Poria cocos using near-infrared spectroscopy combined with chemometrics[J].Spectrochimica Acta Part A, Molecular and Biomolecular Spectroscopy, 2020, 240:118623.
[13] LI X, MA L, ZHANG L.Molecular basis for Poria cocos mushroom polysaccharide used as an antitumor drug in China[J]. Progress in Molecular Biology and Translational Ccience, 2019, 163:263-296.
[14] KIM J H, SIM H A, JUNG D Y, et al.Poria cocus wolf extract ameliorates hepatic steatosis through regulation of lipid metabolism, inhibition of ER stress, and activation of autophagy via AMPK activation[J]. International Journal of Molecular Sciences, 2019, 20(19):4801.
[15] LEE S R, LEE S, MOON E, et al.Bioactivity-guided isolation of anti-inflammatory triterpenoids from the sclerotia of Poria cocos using LPS-stimulated RAW264.7 cells[J].Bioorganic Chemistry, 2017, 70:94-99.
[16] ZHAO J, NIU X, YU J, et al.Poria cocos polysaccharides attenuated ox-LDL-induced inflammation and oxidative stress via ERK activated Nrf2/HO-1 signaling pathway and inhibited foam cell formation in VSMCs[J]. International Immunopharmacology, 2020, 80:106173.
[17] 陈凯琴, 魏科, 叶纯, 等.茯苓多糖对环孢菌素A所致的肺损伤小鼠的免疫调节作用[J].数学中医药, 2022, 5(2):222-232. CHEN K Q, WEI K, YE C, et al.Immunomodulatory effect of pachymaran on cyclosporine A (CsA)-induced lung injury in mice[J].Digital Chinese Medicine, 2022, 5(2):222-232.(in Chinese)
[18] JIN L, GUO Y, MAO W, et al.Total glucosides of paeony inhibit breast cancer growth by inhibiting TAMs infiltration through NF-κB/CCL2 signaling[J].Phytomedicine, 2022, 104:154307.
[19] 刘玉龙, 孙敏, 刘科, 等.丹皮酚通过TLR4/MAPK/NF-κB通路对LPS诱导的巨噬细胞RAW264.7损伤的保护作用[J].海南医学院学报:2022, 29(3):183-189. LIU Y L, SUN M, LIU K, et al Protective effects of paeonol on LPS-induced macrophage RAW264.7 injury through TLR4/MAPK/NF-κB pathway[J]. Journal of Hainan Medical College, 2022, 29(3):183-189.(in Chinese)
[20] YE H, MA S, QIU Z, et al.Poria cocos polysaccharides rescue pyroptosis-driven gut vascular barrier disruption in order to alleviates non-alcoholic steatohepatitis[J].Journal of Ethnopharmacology, 2022, 296:115457.
[21] XU H, WANG S, JIANG Y, et al.Poria cocos polysaccharide aameliorated antibiotic-associated diarrhea in mice via regulating the homeostasis of the gut mmicrobiota and intestinal mucosal barrier[J].International Journal of Molecular Sciences, 2023, 24(2):1423.
[22] RAO K M.Molecular mechanisms regulating iNOS expression in various cell types[J].Journal of Toxicology and Environmental Health-Part B:Critical Reviews, 2000, 3(1):27-58.
[23] GUZIK T J, KORBUT R, ADAMEK-GUZIK T.Nitric oxide and superoxide in inflammation and immune regulation[J].Journal of Physiology and Pharmacology, 2003, 54(4):469-487.
[24] RAWLINGSON A.Nitric oxide, inflammation and acute burn injury[J].Burns, 2003, 29(7):631-640.
[25] ZHU Y, ZHAO Q, HUANG Q, et al.Nuciferine regulates immune function and gut microbiota in DSS-induced ulcerative colitis[J].Frontiers in Veterinary Science, 2022, 9:939377.
[26] JEONG J W, LEE H H, HAN M H, et al.Ethanol extract of Poria cocos reduces the production of inflammatory mediators by suppressing the NF-kappaB signaling pathway in lipopolysaccharide-stimulated RAW264.7 macrophages[J]. BMC Complementary Medicine and Therapies, 2014, 14:101.
[27] BRVNE B.Nitric oxide:NO apoptosis or turning it ON?[J].Cell Death Differ, 2003, 10(8):864-869.
[28] VAN OPDENBOSCH N, LAMKANFI M.Caspases in cell death, inflammation, and disease[J].Immunity, 2019, 50(6):1352-1364.
[29] PARK J, KIM H D, LEE S H, et al.Ascochlorin induces caspase-independent necroptosis in LPS-stimulated RAW264.7 macrophages[J].Journal of Ethnopharmacology, 2019, 239:111898.
[30] VAN LOO G, BERTRAND M J M.Death by TNF:A road to inflammation[J].Nature Reviews Immunology, 2023, 23(5):289-303.
[31] 邵永斌, 郑一闯, 张竞怡, 等.益母草碱对LPS诱导小鼠子宫内膜炎的抗炎作用研究[J].中国畜牧兽医, 2023, 50(5):2149-2155. SHAO Y B, ZHENG Y C, ZHANG J Y, et al.Anti-inflammatory effect of leonuridine on endometritis induced by LPS in mice[J].China Animal Husbandry & Veterinary Medicine, 2023, 50(5):2149-2155.(in Chinese)
[32] WEN L, SUN W, XIA D, et al.The m⁶A methyltransferase METTL3 promotes LPS-induced microglia inflammation through TRAF6/NF-κB pathway[J]. Neuroreport, 2022, 33(6):243-251.
[33] 王巍, 李春洙, 王沙沙, 等.中药联合益生菌对大肠杆菌致小鼠腹泻保护机制研究[J].中国畜牧兽医, 2023, 50(2):779-788. WANG W, LI C Z, WANG S S, et al.Study on the protective mechanism of traditional Chinese medicine combined with probiotics against Escherichia coli diarrhea in mice[J].China Animal Husbandry & Veterinary Medicine, 2023, 50(2):779-788.(in Chinese)
[34] 刘若楠, 邹东敏, 刘茂军, 等.茯苓多糖对磷酸铵镁致羊肾小管上皮细胞损伤的保护作用[J].中国兽医学报, 2022, 42(2):294-298. LIU R N, ZOU D M, LIU M J, et al.Protective effects of Poria cocos polysaccharide extract on injury of sheep renal tubular epithelial cells induced by magnesium ammonium phosphate[J].Chinese Journal of Veterinary Medicine, 2022, 42(2):294-298.(in Chinese)
[35] BAKER S J, REDDY E P.Modulation of life and death by the TNF receptor superfamily[J].Oncogene, 1998, 17(25):3261-3270.
[36] ZHU Z, GAO S, CHEN C, et al.The natural product salicin alleviates osteoarthritis progression by binding to IRE1α and inhibiting endoplasmic reticulum stress through the IRE1α-IκBα-p65 signaling pathway[J].Experimental & Molecular Medicine, 2022, 54(11):1927-1939.
[37] MENG C, WANG S, WANG X, et al.Amphiregulin inhibits TNF-α-induced alveolar epithelial cell death through EGFR signaling pathway[J].Biomedicine & Pharmacotherapy, 2020, 125:109995.
[38] FALEIRO L, LAZEBNIK Y.Caspases disrupt the nuclear-cytoplasmic barrier[J].Journal of Cell Biology, 2000, 151(5):951-959.
[39] 钟粤韵, 杨舒展, 陈非玥, 等.LPS通过ROS/p38/MAPK信号通路对鹅胚肝细胞凋亡的影响[J].中国畜牧兽医, 2023, 50(6):2224-2232. ZHONG Y Y, YANG S Z, CHEN F Y, et al.Effects of LPS on apoptosis of goose embryo hepatocytes through ROS/p38/MAPK signaling pathway[J].China Animal Husbandry & Veterinary Medicine, 2023, 50(6):2224-2232.(in Chinese)
[40] ASHKENAZI A, FAIRBROTHER W J, LEVERSON J D, et al.From basic apoptosis discoveries to advanced selective BCL-2 family inhibitors[J].Nature Reviews Drug discovery, 2017, 16(4):273-284.
[41] TEIJIDO O, DEJEAN L.Upregulation of Bcl-2 inhibits apoptosis-driven Bax insertion but favors BAX relocalization in mitochondria[J].FEBS Letters, 2010, 584(15):3305-3310.
[42] 江文康, 罗冬章, 赵明明, 等.猫脂肪间充质干细胞及其条件培养基对庆大霉素致损猫肾细胞增殖和凋亡的影响[J].中国畜牧兽医, 2021, 48(7):2644-2652. JIANG W K, LUO D Z, ZHAO M M, et al.Effects of cat AD-MSCs and its conditioned medium on apoptosis and proliferation of CRFK damaged by gentamicin[J].China Animal Husbandry & Veterinary Medicine, 2021, 48(7):2644-2652.(in Chinese)
[43] 刘毅.Icariin通过下调NF-κB信号通路降低TNF-α诱导的软骨细胞凋亡和自噬抑制[D].武汉:华中科技大学, 2019. LIU Y.Icariin attenuated the TNF-α induced chondrocyte apoptosis and autophagy inhibition through down regulating NF-κB signal pathway[D].Wuhan:Huazhong University of Science and Technology, 2019.(in Chinese)
[44] 李琪琪, 王雪娇, 初英杰, 等.中国被毛孢醇提物通过MAPK/NF-κB通路双向调控巨噬细胞的免疫功能研究[J].中草药, 2023, 54(11):3559-3567. LI Q Q, WANG X J, CHU Y J, et al.Hirsutella sinensis alcohol extract regulates immune function of macrophages through MAPK/NF-κB pathway pathway[J].Chinese Traditional and Herbal Drugs, 2023, 54(11):3559-3567.(in Chinese)
[45] 聂佳, 赵家宁, 吴甜甜, 等.和厚朴酚对铜绿假单胞菌肺炎小鼠炎症因子及TLR4/MyD88/NF-κB通路的影响[J].广州中医药大学学报, 2023, 40(6):1476-1481. NIE J, ZHAO J N, WU T T, et al.Effects of honokiol on inflammatory factors and TLR4/MyD88/NF-κB pathway in mice with Pseudomonas aeruginosa pneumonia[J].Journal of Guangzhou University of Traditional Chinese Medicine, 2023, 40(6):1476-1481.(in Chinese)
[46] 李宏生, 刘雅林, 孙志敏, 等.IL-1β及NF-κB信号通路在颅内感染大鼠氧化应激反应中的作用及意义[J].中华医院感染学杂志, 2023, 33(7):971-975. LI H S, LIU Y L, SUN Z M et al.Role and significance of IL-1β and NF-κB signaling pathways in oxidative stress in rats with intracranial infection[J].Chinese Journal of Nosocomiology, 2023, 33(7):971-975.(in Chinese)
[47] 邹闻书, 尹朋, 金娜, 等.α-倒捻子素通过抑制TLR4/NF-κB信号通路缓解LPS诱导IEC-6细胞的炎症[J].畜牧兽医学报, 2019, 50(2):431-438. ZOU W S, YIN P, JIN N et al.α-mangostin alleviates LPS-induced inflammation by inhibiting TLR4/NF-κB signaling[J].Journal of Animal Husbandry and Veterinary Medicine, 2019, 50(2):431-438.(in Chinese)
[48] 迟晓晨, 曹瀛心, 包翠芬, 等.人参皂苷Rg1通过TLR4/MyD88/NF-κB p65通路调控小鼠急性肾损伤诱导的急性肝损伤的机制研究[J].中国病理生理杂志, 2023, 39(2):287-296. CHI X C, CAO Y X, BAO C F, et al.Mechanism of ginsenoside Rg1 regulating acute hepatic damage induced by acute kidney injury in mice through TLR4/MyD88/NF-κB p65 signaling pathway[J] Chinese Journal of Pathophysiology, 2023, 39(2):287-296.(in Chinese)
[49] 彭海萍, 吴广礼.山莨菪碱对顺铂诱导的急性肾损伤大鼠TLR4/NF-κB信号通路的调节作用研究[J].中国中西医结合肾病杂志, 2021, 22(9):769-773. PENG H P, WU G L.Regulatory effect of anisodamine on TLR4/NF-κB signal pathway in cisplatin-induced acute renal injury in rats[J].Chinese Journal of Integrative Medicine Nephrology, 2021, 22(9):769-773.(in Chinese)
[50] JHA A K, GAIROLA S, KUNDU S, et al.Toll-like receptor 4:An attractive therapeutic target for acute kidney injury[J].Life Sciences, 2021, 271:119155.
[51] BADRICHANI A Z, STROKA D M, BILBAO G, et al.Bcl-2 and Bcl-XL serve an anti-inflammatory function in endothelial cells through inhibition of NF-kappaB[J].Journal of Clinical Investigation, 1999, 103(4):543-553.

Protective Effect of Poria cocos Polysaccharides on LPS-induced CRFK Cell Inflammation

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	CHEN Yuxiao, FU Haining, CHEN Ling, LUAN Mingna, ZHANG Huimei, YAO Deyong, WANG Yanling, CAI Qingxiu, ZHANG Anyuan, YIN Lingling, SUN Yalei. Study on the Anti-inflammatory,Analgesic and Antipyretic Effects of Erding Extract Powder [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(9): 3861-3868.
[2]	YU Ziyu, ZHANG Bingqi, CAI Yuanrong, HONG Longsheng, LI Wanyan, CAO Nan, HUANG Yunmao, XU Danning, TIAN Yunbo, CHEN Wenbin, LI Bingxin. Screening of Genes Related to Bursa of Fabricius Injury Induced by LPS in Magang Geese [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(7): 2639-2650.
[3]	WU Kaihui, ZHOU Chengli, ZHAO Lingjun, XU Shiyuan, LIU Songqi, DONG Zhihao, YUAN Kaimin, WANG Dong. Research Progress on the Mechanism and Regulatory Pathway of Testicular Injury Induced by Busulfan in Mice [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(7): 2811-2819.
[4]	SUN Weixiang, ZONG Yingying, LIU Bo, ZHUANG Yuan, QIN Feng, CHEN Yu, ZHU Shanyuan, XU Huiqin. Catalpol Alleviates the Damaging Effect of AGEs on the Endothelial Cells in the Hepatic Sinus by Disrupting the Interaction Between Galectin-3 and CD146 [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(7): 2820-2831.
[5]	FAN Qiuyu, WU Jianwen, LI Chunxiao, WANG Jinquan, LI Huanrong, WANG Xiumin. Network Pharmacology Analysis of Anti-inflammatory Mechanism of Sophora alopecuroides and Experimental Validation [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(7): 2951-2965.
[6]	ZHONG Yueyun, YANG Shuzhan, CHEN Feiyue, LU Zhier, LI Bingxin, LI Wanyan, TIAN Yunbo, HUANG Yunmao, XU Danning, CAO Nan. Effects of LPS on Apoptosis of Goose Embryo-derived Hepatocytes ROS/p38/MAPK Signaling Pathway [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2224-2232.
[7]	LI Can, LI Chuchu, ZENG Wei, ZHANG Ning, JI Chunxiao, CHEN Tao. Recombinant Expression and Function Study of RegⅢγ Protein in Porcine [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2414-2426.
[8]	ZHANG Jinwu, WEI Zi, JI Yutong, WU Changxu, LI Zhenmin, WEI Yingyi, HU Tingjun, YU Meiling. Study on Oxidative Stress and Inflammatory Injury in Intestinal Tract of Yellow-feathered Chickens Induced by Clostridium perfringens Type A [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2069-2081.
[9]	SHAO Yongbin, ZHENG Yichuang, ZHANG Jingyi, WEI Linying, TIAN Yong, SUN Guojie, LIU Yuran, LUO Yan, GU Xinli. Study on the Anti-inflammatory Effect of Leonurine on LPS-induced Endometritis in Mice [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2149-2155.
[10]	HU Honglian, LI Chao, CHEN Liqing, SONG Liwen, ZHAO Meng, ZHANG Jianxia, WANG Dian, GAO Min. Effects of Dietary β-carotene Supplementation on Lactation Performance, Blood Routine,Serum Biochemical and Immune Indexes of Dairy Cows during the Lactating Peak Period [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1375-1384.
[11]	GUO Yaru, LI Kexin, WANG Li, LIU Jinyu, SHI Lei, REN Youshe. Effects of Selenium on Lead-Induced Apoptosis of Mouse Leydig Cells in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1404-1411.
[12]	HUANG Xiaoqian, YUAN Junzhao, LIU Jinhe, ZOU Hui, GU Jianhong, YUAN Yan, LIU Xuezhong, LIU Zongping, BIAN Jianchun. Protective Effect of Honokiol on Cadmium-induced Renal Injury in Chickens [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1007-1015.
[13]	FU Jingcheng, HE Junhui, TONG Jiang, WU Wei, GUO Shuang, YANG Yanbin, HAN Yingqian, WANG Yueying, LI Heping. Anti-inflammatory and Antioxidant Roles of HO-1 in Macrophages [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 807-816.
[14]	TAO Yanzi, WANG Qiuju, YUAN Ling, CUI Yizhe, JIA Junfeng. Effect of LPS-induced Inflammatory Time on IAP and NF-κB Expression in Small Intestine of Mice [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(11): 4424-4432.
[15]	MA Xiaolan, DONG Zhen, YUE Meishan, CHANG Guanyu, ZENG Jianguo. Effect of Macleaya cordata Leaf Extract on Ulcerative Colitis in Mice [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(11): 4664-4680.