37 2 Vol37 No2 2018 6 Journal of South-Central University for NationalitiesNatSciEdition Jun2018 1 1 2 1 1 1 1 4300742 430074 MTT 200 μg ml -1 ConA 700% ConA IL-6 TNF-α 150 mg kg -1 790%ConA 433% 1483% ConA 485% Q956 A 1672-4321201802-0049-05 Effects of Polygonatum sibiricum Polysaccharide on Immunosuppressive Action Induced by Cyclophosphamide in Mice Deng Xukun 1 Duan Huan 1 Liu Zhao 2 Shu Guangwen 1 Jiang Shanqing 1 Yu Huifan 1 1 College of Pharmaceutical SciencesSouth-Central University for NationalitiesWuhan 430074China 2 Editorial Department of University Journal South-Central University for Nationalities Wuhan 430074 China Abstract ObjectiveTo investigate the effects of Polygonatum sibiricum polysaccharidepspon the immunosuppression activities induced by cyclophosphamide in mice MethodsMTT assay was used to evaluate the proliferation activity of splenocytes of normal mice Mice immunosuppression model was established with cyclophosphamide The mice spleen thymus index and spleen cellthymocyte proliferation activity in vitromacrophage phagocytosisand the ability of macrophages to secrete inflammatory cytokines were respectively determined ResultPSP could evidently increase the proliferation activity of spleen cell of normal mice The synergistic proliferation promotion rate with ConA was more than 700% with PSP dose of 200 μg ml -1 PSP could also improve the spleen and thymus index of immunosuppressed mice promote the proliferation of spleen and thymus cells stimulated by ConAand enhance the phagocytosis of macrophage and the secretion of IL-6 and TNF-α Compared with the control group the spleen index increased by 790%433% in high PSP dose 150 mg kg -1 for ConA synergized groupand the thymus indexes increased by 1483%485% respectively ConclusionPSP can significantly alleviate the immunosuppression of mice induced by cyclophosphamide Keywords Polygonatum sibiricum polysaccharidecyclophosphamideimmunosuppression Polygonatum 1 kingianum Coll et Hemsl Polygonatum sibiricum Red Polygonatum cyrtonema Hua 2017-12-25 2 1972- E-maildxk711208@ 163com 81073147 2012BAI27B06
50 37 22 3 50 5 50100150 mg kg -1 IL-2 4 10 001 ml g -1 2 mg 3 d 4 d 1 1 23 5 ml RPMI-1640 11 15 min 7 ~ 9 SPF 18 ~ 22 g 2 RPMI-1640 SCXK2016-0003 3 d 95% 12 6 Polygonatum = mg/ g sibiricum Red RPMI-1640 RPMI-1640 Genom Tissue Culture Biologicals MTT ConA LPSSigma IL- 1β IL-6 TNF-α 25 13 Spectra Max M3 Dynex CO 2 BB15 Thermo BL- 600 4 h 3 Suprafuge22 Heraeus 200 μl - 1 1 RE52CS-1 24 / 200 083% NH 4 Cl ph 72 96 2 10 5 /37 5% CO 2 4 h PBS 100 μl 0 04% 12 h OD 570 2 26 ConA 21 100 g 95% 96 5 10 5 /24 h 60 3 h 80% 60 2 h 100 μl ConA2 μg ml -1 50 RPMI-1640 72 h 10 μl / 1 25 80 3 MTT 4 h 150 μl 3 h 80% DMSO OD 570 ConA 4 = OD / 48 h OD 7-27 IL-1βIL-6TNF-α 969% 6 24 1 10 6 /
2 51 20 μg ml -1 LPS24 h t x± 珋 s P<005 ELISA IL-1βIL-6 TNF-α 28 96 2 10 5 / 6 ConA 02550100 ConA 200400 μg /ml ConA 1 1 50 μg ml -1 ConA 10 μl 02550100200400 OD 570 OD 150 mg kg -1 29 SPSS 190 P<001 3 31 391% ConA 309% P < 005 μg ml -1 48 h 10 μl MTT 4 h 150 μl DMSO ConA 790% P <001ConA 433%P<005 Tab1 1 ConA x±sn 珋 = 10 Effects of PSP on spleen index and proliferation ofsplenocyte stimulated by ConA in immunosuppression mice /mg kg -1 / mg g -1 ConA 399±017 646±049 243±037 446±019 50 258±013 541±029 100 414±029 * 582±068 150 435±026 ** 639±081 * P<005 * ** P<001 32 ConA ConA 150 mg kg -1 2 2 1483%P<001ConA ConA 485%P<005 522%345%P<005 Tab2 2 ConA x±sn 珋 = 10 Effects of PSP on thymus index and proliferation ofthymocyte stimulated by ConA in immunosuppression mice /mg kg -1 / mg g -1 ConA 299±026 664±049 143±032 435±029 50 218±023 445±019 100 314±027 * 528±056 150 355±039 ** 646±083 * P<005 * ** P<001
52 37 1βIL-6 TNF-α 33 IL- TNF-α P <005 IL- 1β IL-6TNF-α 3 3 IL-6 P<001 Tab3 3 IL-6 TNF-α 150 mg kg -1 IL-6 TNF-α IL-1βIL-6TNF-α x±sn 珋 = 10 Effects of PSP on the secretion of IL-1βIL-6TNF-α in macrophages of immunosuppression mice /mg kg -1 / pg ml -1 IL-1β IL-6 TNF-α 3374±338 3577±461 17809±241 3268±347 3074±410 14663±407 50 3358±345 3489±456 17421±237 100 3387±348 3657±475 * 17863±642 * 150 3454±353 * 898±501 ** 18018±343 ** P<005 P<001 * P<005 ** P<001 34 4 1 1 P<001 P<005 1 P<001 * P<005 Fig1 Effect of PSP on phagocytosis of macrophages in immunosuppression mice ConA 35 4 4 ConA ConA 200400 μg ml -1 9 P < 10 0001 ConA 760% Tab4 珋 x±sn =6 Effect of PSP on the proliferation activity of spleen cells in normal mice / μg ml -1 ConA+ 0 100±013 142±010 25 109±035 183±019 50 124±012 357±037 * 100 176±030 * 414±043 ** 200 219±057 ** 739±076 *** 400 432±087 *** 760±105 *** * P<005 ** P<001 *** P<0001 4 T B 8
2 53 TNF-αIL-1β IL-2 IL-6 J 2004 19312-13 8 11 LPS 5 J IL-6 TNFα 6 2017 363 52-56 J 201635 2141-143 7 ConA J 2016 IL-6 TNF-α 351 67-69 8 J 2012 283112-114 9 J 2016 39 1183-187 1 M 10Schepetkin I AQuinn M T Botanical polysaccharides 2015138-139 macrophage immunomodulation and therapeutic potential 2 J J Int Immunopharmacol2006 63 317-333 20143196-199 11Liu J Qi YZheng Let al Xinfeng capsule improves 3 pulmonary function in ankylosing spondylitis patients via J 2011 34 NF-κB-iNOS-NO signaling pathway J J Tradit Chin 191-94 Med2014346 657-665 4 櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖櫖 19 LiNi 1/3 Co 1/3 Mn 1/3 O 2 powders cathode material for Li-ion batteries J J Power Sources 2010 195103047-52 1Florian S Prasant K N Evan M E et al Study of cathode materials for lithium-ion batteriesrecent progress and new challenges J Inorganics 2017 5232-41 2Yuan X Xu Q J Liu X et al Excellent rate performance and high capacity of Mo doped layered cathode material Li Li 02 Mn 054 Ni 013 Co 013 O 2 derived from an improved coprecipitation approach J Electrochim Acta 2016 207 120-129 3 CeO 2 J 2016 353 17-20 4Rosedhi N D Idris N H Rahman M M et al Disordered spinel LiNi 05 Mn 15 O 4 cathode with improved rate performance for lithium-ion batteries J Electrochim Acta 2016 206374-380 5Han X Meng Q Sun T et al Preparation and electrochemical characterization of single-crystalline spherical 6Zhang S Deng C Fu B L et al Synthetic optimization of spherical LiNi 1/3 Mn 1/3 Co 1/3 O 2 prepared by a carbonate co-precipitation method J Powder Technol 2010 198 3373-380 7Wang T Liu Z H Fan L et al Synthesis optimization of Li 1+x Mn 045 Co 040 Ni 015 O 2 with different spherical sizes via co-precipitation J Powder Technol 2008 1872 124-129 8Zheng Z Guo X D Chou S L et al Uniform Ni-rich LiNi 06 Co 02 Mn 02 O 2 porous microspheresfacile designed synthesis and their improved electrochemical performance J Electrochim Acta 2016 191401-410 9Ellis B Subramanya Herle P Rho Y H et al Nanostructured materials for lithium-ion batteriessurface conductivity vs bulk ion/electron transport J Faraday Discuss 2007 134 119-141