白藜芦醇调控Th17/Treg抑制血吸虫病肝脏肉芽肿
|
摘要
目的探讨白藜芦醇对血吸虫病肝脏肉芽肿作用,分析其对Th17、Treg应答的影响。方法 45只C57BL/6小鼠感染日本血吸虫3周后,随机分为感染组、灌胃白藜芦醇治疗组、灌胃吡喹酮治疗组,另取15只正常小鼠为健康对照组。感染第9周,取肝脏HE染色观察虫卵肉芽肿,计数中性粒细胞;RT-PCR检测肝脏中IL-17A、Foxp3、CXCL1、CXCL2水平;流式细胞术检测脾脏中Th17、Treg应答。用PBS、SEA刺激正常小鼠脾脏细胞,分别加入白藜芦醇和吡喹酮,流式细胞术检测Th17、Treg应答。结果与感染组相比,白藜芦醇治疗组,肝脏肉芽肿减轻,肉芽肿中性粒细胞数量减少(P<0.01),Th17应答减弱(P<0.05),Treg应答增强(P<0.05),肝脏IL-17A、CXCL1、CXCL2表达均降低(P<0.05),Foxp3表达增高(P<0.01)。白藜芦醇促使SEA刺激的T细胞往Th17细胞分化减少(P<0.05),往Treg细胞分化增多(P<0.05)。结论白藜芦醇通过降低Th17应答,增强Treg应答,抑制血吸虫病肝脏肉芽肿。
Aim To observe the anti-granuloma of resveratrol in mice with schistosomiasis japonica and its effect on Th17 and Treg responses. Methods Forty-five mice infected with S. japonicum cercariae for three weeks were randomly divided into three groups named as infected group(A), resveratrol group(B) and praziquantel group(C). Fifteen normal mice were taken as normal control group(D). At the 9 th week post-infection, the granuloma of the liver tissues was observed using HE staining, and the number of neutrophils was calculated. IL-17 A, CXCL1, CXCL2, Foxp3 levels in liver tissues were detected by RT-PCR. Th17 and Treg responses in spleen were determined by FCM respectively. Splenocytes from healthy mice were stimulated by PBS and SEA respectively. After SEA stimulation, resveratrol and praziquantel were added respectively, and Th17 and Treg responses were detected by FCM. Results Compared to group A, the granuloma was alleviated(P<0.01), the number of neutrophils in single granuloma was reduced(P<0.01), Th17 response significantly decreased(P<0.05) and Treg response increased markedly in group B(P<0.05). The hepatic expression of IL-17 A, CXCL1, CXCL2 levels in group B was lower than that of group A(P<0.05), and Foxp3 level in group B were higher than in group A(P<0.01). In vitro, resveratrol inhibited the differentiation of Th17 cells(P<0.05) and enhanced that of Treg cells after SEA treatment(P<0.05). Conclusion Resveratrol inhibits the heptic granulomatous response through down-regulating Th17 response and up-regulating Treg response in mice with schistosomiasis.
Aim To observe the anti-granuloma of resveratrol in mice with schistosomiasis japonica and its effect on Th17 and Treg responses. Methods Forty-five mice infected with S. japonicum cercariae for three weeks were randomly divided into three groups named as infected group(A), resveratrol group(B) and praziquantel group(C). Fifteen normal mice were taken as normal control group(D). At the 9 th week post-infection, the granuloma of the liver tissues was observed using HE staining, and the number of neutrophils was calculated. IL-17 A, CXCL1, CXCL2, Foxp3 levels in liver tissues were detected by RT-PCR. Th17 and Treg responses in spleen were determined by FCM respectively. Splenocytes from healthy mice were stimulated by PBS and SEA respectively. After SEA stimulation, resveratrol and praziquantel were added respectively, and Th17 and Treg responses were detected by FCM. Results Compared to group A, the granuloma was alleviated(P<0.01), the number of neutrophils in single granuloma was reduced(P<0.01), Th17 response significantly decreased(P<0.05) and Treg response increased markedly in group B(P<0.05). The hepatic expression of IL-17 A, CXCL1, CXCL2 levels in group B was lower than that of group A(P<0.05), and Foxp3 level in group B were higher than in group A(P<0.01). In vitro, resveratrol inhibited the differentiation of Th17 cells(P<0.05) and enhanced that of Treg cells after SEA treatment(P<0.05). Conclusion Resveratrol inhibits the heptic granulomatous response through down-regulating Th17 response and up-regulating Treg response in mice with schistosomiasis.
引文
[1] Pearce E J, MacDonald A S. The immunobiology of schistosomiasis[J]. Nat Rev Immunol, 2002,2(7):499-511.
[2] Kaplan M H, Whitfield J R, Boros D L,et al. Th2 cells are required for the Schistosoma mansoni egg-induced granulomatous response[J]. J Immunol, 1998,160(7): 1850-6.
[3] Stadecker M J, Asahi H, Finger E. The immunobiology of Th1 polarization in high-pathology schistosomiasis[J]. Immunol Rev, 2004, 201(1):168-79.
[4] Chen D, Luo X, Xie H, et al. Characteristics of IL-17 induction by Schistosoma japonicum infection in C57BL/6 mouse liver[J]. Immunology, 2013,139(4): 523-32.
[5] Turner J D, Jenkins G R, Hogg K G, et al. CD4+CD25+ regulatory cells contribute to the regulation of colonic Th2 granulomatous pathology caused by schistosome infection[J]. PLoS Negl Trop Dis, 2011,5(8): e1269.
[6] 王坦,张艳群,曾永联,等.白藜芦醇抑制人肝癌SMMC-7721细胞增殖并降低mTOR蛋白磷酸化水平[J].中国药理学通报, 2017,33(9):1309-14.
[6] Wang T, Zhang Y Q, Zeng Y L,et al. Effect of resveratrol on proliferation of liver cancer SMMC-7721 cells and lowering levels of mTOR protein phosphorylation[J].Chin Pharmacol Bull, 2017,33(9):1309-14.
[7] 陈淳,林建伟,李国平,等.白藜芦醇调控大鼠急性肺栓塞后肺动脉高压及MCP-1表达的机制研究[J].中国药理学通报, 2017,33(10):1436-41.
[7] Chen C,Lin J W,Li G P, et al. Resveratrol down-regulates acute pulmonary thromboembolism-induced pulmonary artery hypertension and monocyte chemoattractant protein-1 in rats[J].Chin Pharmacol Bull, 2017,33(10):1436-41.
[8] 张伟伟,朱继峰,王任,等.白藜芦醇调控Th1和Th2应答抑制小鼠日本血吸虫病肝脏纤维化的研究[J]. 中国药理学通报, 2016,32(8):1091-7.
[8] Zhang W W,Zhu J F,Wang R, et al. The study on resveratrol inhibited hepatic fibrosis in mice with schistosomiasis japonica by modulating Th1 and Th2 responses[J].Chin Pharmacol Bull, 2016,32(8):1091-7.
[9] Yao J, Wei C, Wang J Y, et al. Effect of resveratrol on Treg/Th17 signaling and ulcerative colitis treatment in mice[J]. World J Gastroenterol, 2015,21(21):5672-81.
[10] 钟淼,郭颖,邓建云,等.异丹叶大黄素与白藜芦醇对兔外周血中性粒细胞功能的影响[J]. 药学学报, 1998,33(11):13-6.
[10] Zhong M,Guo Y,Deng J Y,et al. Effects of isorhapotigenin and resveratrol on function of peripheral blood polymorphonuclear leukocytes from rabbits [J].Acta Pharm Sin, 1998,33(11):13-6.
[11] Zhou L, Lopes J E, Chong M M, et al. TGF-beta-induced Foxp3 inhibits Th17 cell differentiation by antagonizing RORgamma t function[J]. Nature, 2008,453(7192): 236-40.
[12] Yasumi Y, Takikawa Y, Endo R, et al.Interleukin-17 as anew marker of severity of acute hepatic injury[J]. Hepatol Res, 2007,37(4): 248-54.
[13] Cong Y, Konrad A, Iqbal N, et al. Generation of antigen-specific,Foxp3-expressing CD4+ regulatory T cells by inhibition of APC proteosome function[J]. J Immunol, 2005,174(4): 2787-95.
[14] Shi G, Lovaas J D, Tan C, et al. Cell-cell interaction with APC, not IL-23, is required for naive CD4 cells to acquire pathogenicity during Th17 lineage commitment[J]. J Immunol, 2012,189(3): 1220-7.
[15] Imler T J Jr,Petro T M. Decreased severity of experimental autoimmune encephalomyelitis during resveratrol administration is associated with increased IL-17+IL-10+ T cells, CD4(-) IFN-gamma+ cells, and decreased macrophage IL-6 expression[J]. Int Immunopharmacol, 2009,9(1):134-43.
[2] Kaplan M H, Whitfield J R, Boros D L,et al. Th2 cells are required for the Schistosoma mansoni egg-induced granulomatous response[J]. J Immunol, 1998,160(7): 1850-6.
[3] Stadecker M J, Asahi H, Finger E. The immunobiology of Th1 polarization in high-pathology schistosomiasis[J]. Immunol Rev, 2004, 201(1):168-79.
[4] Chen D, Luo X, Xie H, et al. Characteristics of IL-17 induction by Schistosoma japonicum infection in C57BL/6 mouse liver[J]. Immunology, 2013,139(4): 523-32.
[5] Turner J D, Jenkins G R, Hogg K G, et al. CD4+CD25+ regulatory cells contribute to the regulation of colonic Th2 granulomatous pathology caused by schistosome infection[J]. PLoS Negl Trop Dis, 2011,5(8): e1269.
[6] 王坦,张艳群,曾永联,等.白藜芦醇抑制人肝癌SMMC-7721细胞增殖并降低mTOR蛋白磷酸化水平[J].中国药理学通报, 2017,33(9):1309-14.
[6] Wang T, Zhang Y Q, Zeng Y L,et al. Effect of resveratrol on proliferation of liver cancer SMMC-7721 cells and lowering levels of mTOR protein phosphorylation[J].Chin Pharmacol Bull, 2017,33(9):1309-14.
[7] 陈淳,林建伟,李国平,等.白藜芦醇调控大鼠急性肺栓塞后肺动脉高压及MCP-1表达的机制研究[J].中国药理学通报, 2017,33(10):1436-41.
[7] Chen C,Lin J W,Li G P, et al. Resveratrol down-regulates acute pulmonary thromboembolism-induced pulmonary artery hypertension and monocyte chemoattractant protein-1 in rats[J].Chin Pharmacol Bull, 2017,33(10):1436-41.
[8] 张伟伟,朱继峰,王任,等.白藜芦醇调控Th1和Th2应答抑制小鼠日本血吸虫病肝脏纤维化的研究[J]. 中国药理学通报, 2016,32(8):1091-7.
[8] Zhang W W,Zhu J F,Wang R, et al. The study on resveratrol inhibited hepatic fibrosis in mice with schistosomiasis japonica by modulating Th1 and Th2 responses[J].Chin Pharmacol Bull, 2016,32(8):1091-7.
[9] Yao J, Wei C, Wang J Y, et al. Effect of resveratrol on Treg/Th17 signaling and ulcerative colitis treatment in mice[J]. World J Gastroenterol, 2015,21(21):5672-81.
[10] 钟淼,郭颖,邓建云,等.异丹叶大黄素与白藜芦醇对兔外周血中性粒细胞功能的影响[J]. 药学学报, 1998,33(11):13-6.
[10] Zhong M,Guo Y,Deng J Y,et al. Effects of isorhapotigenin and resveratrol on function of peripheral blood polymorphonuclear leukocytes from rabbits [J].Acta Pharm Sin, 1998,33(11):13-6.
[11] Zhou L, Lopes J E, Chong M M, et al. TGF-beta-induced Foxp3 inhibits Th17 cell differentiation by antagonizing RORgamma t function[J]. Nature, 2008,453(7192): 236-40.
[12] Yasumi Y, Takikawa Y, Endo R, et al.Interleukin-17 as anew marker of severity of acute hepatic injury[J]. Hepatol Res, 2007,37(4): 248-54.
[13] Cong Y, Konrad A, Iqbal N, et al. Generation of antigen-specific,Foxp3-expressing CD4+ regulatory T cells by inhibition of APC proteosome function[J]. J Immunol, 2005,174(4): 2787-95.
[14] Shi G, Lovaas J D, Tan C, et al. Cell-cell interaction with APC, not IL-23, is required for naive CD4 cells to acquire pathogenicity during Th17 lineage commitment[J]. J Immunol, 2012,189(3): 1220-7.
[15] Imler T J Jr,Petro T M. Decreased severity of experimental autoimmune encephalomyelitis during resveratrol administration is associated with increased IL-17+IL-10+ T cells, CD4(-) IFN-gamma+ cells, and decreased macrophage IL-6 expression[J]. Int Immunopharmacol, 2009,9(1):134-43.