滋阴补肾方联合糖皮质激素对系统性红斑狼疮小鼠T淋巴细胞亚群和炎症细胞因子的影响
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摘要
目的:探讨滋阴补肾方联合糖皮质激素对系统性红斑狼疮(SLE)小鼠T淋巴细胞亚群和炎症细胞因子的影响,阐明中、西药联合对SLE的作用机制。方法:将50只MRL/lpr小鼠随机分为模型组,西药组,中药组,低和高剂量联合组(n=10);另选10只C57BL/6小鼠作为对照组。对照组和模型组小鼠灌胃给予0.5mL·d~(-1)生理盐水,西药组小鼠灌胃给予12.5mg·kg~(-1)醋酸泼尼松(糖皮质激素),中药组小鼠灌胃给予25.0g·kg~(-1)滋阴补肾方,低和高剂量联合组小鼠分别灌胃给予12.5和25.0g·kg~(-1)滋阴补肾方和6.25mg·kg~(-1)醋酸泼尼松。连续给药21d后,计算各组小鼠脾脏指数和胸腺指数。采用流式细胞术检测各组小鼠外周血中CD3~+、CD4~+和CD8~+T淋巴细胞亚群水平。采用酶联免疫吸附试验(ELISA)法检测小鼠血清白细胞介素4 (IL-4)、白细胞介素6 (IL-6)和γ-干扰素(IFN-γ)水平。结果:与模型组比较,西药组和低及高剂量联合组小鼠脾脏指数和胸腺指数明显降低(P<0.05);与模型组比较,中药组,低和高剂量联合组CD3~+和CD4~+T淋巴细胞水平明显升高(P<0.05);与模型组比较,西药组、中药组和低及高剂量联合组CD8+T淋巴细胞水平明显降低(P<0.05),血清中IL-4和IL-6水平明显降低(P<0.05),IFN-γ水平明显升高(P<0.05),且系膜细胞与系膜基质增生明显改善,肾间质炎性细胞浸润减轻;与低剂量联合组比较,高剂量联合组小鼠CD4~+T淋巴细胞水平、CD4~+/CD8~+比值和IFN-γ水平明显升高(P<0.05),CD8~+T淋巴细胞水平明显降低(P<0.05),其他指标比较差异均无统计学意义(P>0.05)。结论:滋阴补肾方联合糖皮质激素可降低SLE小鼠炎性细胞因子水平,调节T淋巴细胞亚群平衡,弥补单用糖皮质激素免疫抑制治疗后不能扶正机体内环境的不足,有效调节机体免疫平衡。
Objective:To investigate the effects of Nourishing Kidney-Yin Prescription combined with glucocorticoid on the T lymphocyte subsets and inflammatory cytokines in the mice with systemic lupus erythematosus(SLE),and to elucidate the mechanism of Chinese and Western medicine combination in the treatment of SLE.Methods:Fifty MRL/lpr mice were randomly divided into model group,Western medicine group,Chinese medicine group,low dose and high dose of combintion groups(n=10).Ten C57 BL/6 mice were selected as control group.The mice in control group and model group were given 0.5 mL·d-1 normal saline by gavage;the mice in Western medicine group were given 12.5 mg·kg~(-1) prednisone acetate(glucocorticoid)by gavage;the mice in Chinese medicine group were given 25.0 g·kg~(-1) Nourishing Kindey-Yin Prescription by gavage;the mice in low and high doses of combination groups were given 12.5 and 25.0 g·kg~(-1) Nourishing Kidney-Yin prescription by gavage and 6.25 mg·kg~(-1) prednisone acetate,respectively.After 21 dof continuous administration,the spleen indexes and thymus indexes of the mice in various groups were calculated.The levels of CD3+,CD4+and CD8+T lymphocyte subsets in the periphreal blood of the mice in various groups were determined by flow cytometry.The serum levels of interleukin-4(IL-4),interleukin-6(IL-6)andγ-interferon(IFN-γ)were measured by enzyme-linked immunosorbent assay(ELISA)method.Results:Compared with model group,the spleen indexes and thymus indexes of the mice in Western medicine,low dose of combination group and high dose of combination group were decreased obviously(P <0.05).Compared with model group,the levels of CD3+lymphocytes and CD4+T lymphocytes in Chinese medicine group,low dose of combintion group and high dose of combination group were significantly increased(P<0.05).Compared with model group,the CD8+T lymphocyte levels in Western medicine group,Chinese medicine group,low dose combination group and high dose combination group were significantly decreased(P<0.05),the serum IL-4 and IL-6 levels were obviously decreased(P<0.05),and the IFN-γlevels were significantly increased(P<0.05);the mesagial cell and mesangial matrix hyperplasia were improved,and the renal intestitial inflammatory cell infiltration was relieved.Conclusion:Nourishing KidneyYin Prescription combined with glucocorticoid can effectively regulate the body's immune balance by reducing the inflammatory cytokines,regulating T lymphocyte subgroup equilibrium,and recovering the deficiency of the immune-suppression therapy by monosaccharide corticosteroids.
Objective:To investigate the effects of Nourishing Kidney-Yin Prescription combined with glucocorticoid on the T lymphocyte subsets and inflammatory cytokines in the mice with systemic lupus erythematosus(SLE),and to elucidate the mechanism of Chinese and Western medicine combination in the treatment of SLE.Methods:Fifty MRL/lpr mice were randomly divided into model group,Western medicine group,Chinese medicine group,low dose and high dose of combintion groups(n=10).Ten C57 BL/6 mice were selected as control group.The mice in control group and model group were given 0.5 mL·d-1 normal saline by gavage;the mice in Western medicine group were given 12.5 mg·kg~(-1) prednisone acetate(glucocorticoid)by gavage;the mice in Chinese medicine group were given 25.0 g·kg~(-1) Nourishing Kindey-Yin Prescription by gavage;the mice in low and high doses of combination groups were given 12.5 and 25.0 g·kg~(-1) Nourishing Kidney-Yin prescription by gavage and 6.25 mg·kg~(-1) prednisone acetate,respectively.After 21 dof continuous administration,the spleen indexes and thymus indexes of the mice in various groups were calculated.The levels of CD3+,CD4+and CD8+T lymphocyte subsets in the periphreal blood of the mice in various groups were determined by flow cytometry.The serum levels of interleukin-4(IL-4),interleukin-6(IL-6)andγ-interferon(IFN-γ)were measured by enzyme-linked immunosorbent assay(ELISA)method.Results:Compared with model group,the spleen indexes and thymus indexes of the mice in Western medicine,low dose of combination group and high dose of combination group were decreased obviously(P <0.05).Compared with model group,the levels of CD3+lymphocytes and CD4+T lymphocytes in Chinese medicine group,low dose of combintion group and high dose of combination group were significantly increased(P<0.05).Compared with model group,the CD8+T lymphocyte levels in Western medicine group,Chinese medicine group,low dose combination group and high dose combination group were significantly decreased(P<0.05),the serum IL-4 and IL-6 levels were obviously decreased(P<0.05),and the IFN-γlevels were significantly increased(P<0.05);the mesagial cell and mesangial matrix hyperplasia were improved,and the renal intestitial inflammatory cell infiltration was relieved.Conclusion:Nourishing KidneyYin Prescription combined with glucocorticoid can effectively regulate the body's immune balance by reducing the inflammatory cytokines,regulating T lymphocyte subgroup equilibrium,and recovering the deficiency of the immune-suppression therapy by monosaccharide corticosteroids.
引文
[1]游弋.系统性红斑狼疮2017年研究进展[J].皮肤病与性病,2018,40(2):176-177.
[2]ALI M,FIROZ C K,JABIR N R,et al.An insight on the pathogenesis and treatment of systemic lupus erythematosus[J].Endocr Metab Immune Disord Drug Targets,2018,18(2):110-123.
[3]王大维,汪梅姣,谷焕鹏,等.从雌激素受体探讨解毒祛瘀滋阴药对SLE的治疗机制[J].中国中医急症,2017,26(9):1505-1508,1513.
[4]韩曼,姜泉.中医治疗系统性红斑狼疮的思路与实践[J].中华中医药杂志,2017,32(10):4537-4539.
[5]郑虹.益气养阴法联合糖皮质激素对系统性红斑狼疮患者免疫功能的影响[J].时珍国医国药,2014,25(4):894-896.
[6]林昌松,李宁,徐强,等.“知柏养阴汤”联合糖皮质激素治疗系统性红斑狼疮60例临床研究[J].江苏中医药,2011,43(11):13-15.
[7]林宁,钟嘉熙,邱斌,等.青蒿鳖甲汤加减对MRL/lpr狼疮鼠Th17细胞及肾脏病理的影响[J].广州中医药大学学报,2014,31(5):776-779,785.
[8]沈家敏,吴艮艮,梁涛.具有SLE疾病症状的MRL/lpr小鼠体内自噬水平检测[J].免疫学杂志,2017,33(10):867-871.
[9]张青华,李南,陈嵘祎,等.T细胞亚群和白细胞介素36在系统性红斑狼疮发病机制中的研究进展[J].国际皮肤性病学杂志,2016,42(3):178-180.
[10]林丽,万春平,李小丝,等.益气养阴补肾方联合激素对自发狼疮转基因小鼠增效减毒作用的研究[J].时珍国医国药,2017,28(5):1044-1046.
[11]张颖,李珍.狼疮性肾炎的中西医护理研究[J].内蒙古中医药,2014,33(16):156-157.
[12]胡笛,程丑夫.程丑夫从胃热论治疑难杂症验案4则[J].湖南中医杂志,2017,33(3):96-98.
[13]KARRAR S,CUNNINGHAME GRAHAM D S.Abnormal B-cell development in systemic lupus erythematosus:what the genetics tell us[J].Arthritis Rheumatol,2018,70(4):496-507.
[14]MIZUI M,TSOKOS G C.Targeting regulatory T cells to treat patients with systemic lupus erythematosus.[J].Front Immunol,2018,9(4):1-9.
[15]ZHOU H Y,HU B L,HUANG N W,et al.Aberrant T cell subsets and cytokines expression profile in systemic lupus erythematosus[J].Clin Rheumatol,2018,37(9):2405-2413.
[16]顾敏,谢雁鸣.中药过敏反应中细胞因子的表达[J].国际检验医学杂志,2016,37(19):2740-2742.
[17]甄洪花,王金良,单虎,等.猪IL-2与IL-6的融合表达及其产物的促淋巴细胞增殖活性[J].畜牧兽医学报,2010,41(6):741-745.
[18]FLEISCHER S,RIES S,SHEN P,et al.Anti-interleukin-6signalling therapy rebalances the disrupted cytokine production of B cells from patients with active rheumatoid arthritis[J].Eur J Immunol,2018,48(1):194-203.
[19]CHASSET F,ARNAUD L.Targeting interferons and their pathways in systemic lupus erythematosus[J].Autoimmun Rev,2018,17(1):44-52.
[20]YAO Y,WANG J B,XIN M M,et al.Balance between inflammatory and regulatory cytokines in systemic lupus erythematosus[J].Genet Mole Res,2016,15(2):1-8.
[21]徐毅芳,寿张飞.调节性T细胞在移植耐受中的作用[J].国际移植与血液净化杂志,2008,6(2):40-43.
[22]葛宽雪,高伟华,向晓星.辅助性T淋巴细胞17和调节性T淋巴细胞及其平衡改变与自身免疫性肝炎的关系[J].临床肝胆病杂志,2018,34(7):1573-1576.
[2]ALI M,FIROZ C K,JABIR N R,et al.An insight on the pathogenesis and treatment of systemic lupus erythematosus[J].Endocr Metab Immune Disord Drug Targets,2018,18(2):110-123.
[3]王大维,汪梅姣,谷焕鹏,等.从雌激素受体探讨解毒祛瘀滋阴药对SLE的治疗机制[J].中国中医急症,2017,26(9):1505-1508,1513.
[4]韩曼,姜泉.中医治疗系统性红斑狼疮的思路与实践[J].中华中医药杂志,2017,32(10):4537-4539.
[5]郑虹.益气养阴法联合糖皮质激素对系统性红斑狼疮患者免疫功能的影响[J].时珍国医国药,2014,25(4):894-896.
[6]林昌松,李宁,徐强,等.“知柏养阴汤”联合糖皮质激素治疗系统性红斑狼疮60例临床研究[J].江苏中医药,2011,43(11):13-15.
[7]林宁,钟嘉熙,邱斌,等.青蒿鳖甲汤加减对MRL/lpr狼疮鼠Th17细胞及肾脏病理的影响[J].广州中医药大学学报,2014,31(5):776-779,785.
[8]沈家敏,吴艮艮,梁涛.具有SLE疾病症状的MRL/lpr小鼠体内自噬水平检测[J].免疫学杂志,2017,33(10):867-871.
[9]张青华,李南,陈嵘祎,等.T细胞亚群和白细胞介素36在系统性红斑狼疮发病机制中的研究进展[J].国际皮肤性病学杂志,2016,42(3):178-180.
[10]林丽,万春平,李小丝,等.益气养阴补肾方联合激素对自发狼疮转基因小鼠增效减毒作用的研究[J].时珍国医国药,2017,28(5):1044-1046.
[11]张颖,李珍.狼疮性肾炎的中西医护理研究[J].内蒙古中医药,2014,33(16):156-157.
[12]胡笛,程丑夫.程丑夫从胃热论治疑难杂症验案4则[J].湖南中医杂志,2017,33(3):96-98.
[13]KARRAR S,CUNNINGHAME GRAHAM D S.Abnormal B-cell development in systemic lupus erythematosus:what the genetics tell us[J].Arthritis Rheumatol,2018,70(4):496-507.
[14]MIZUI M,TSOKOS G C.Targeting regulatory T cells to treat patients with systemic lupus erythematosus.[J].Front Immunol,2018,9(4):1-9.
[15]ZHOU H Y,HU B L,HUANG N W,et al.Aberrant T cell subsets and cytokines expression profile in systemic lupus erythematosus[J].Clin Rheumatol,2018,37(9):2405-2413.
[16]顾敏,谢雁鸣.中药过敏反应中细胞因子的表达[J].国际检验医学杂志,2016,37(19):2740-2742.
[17]甄洪花,王金良,单虎,等.猪IL-2与IL-6的融合表达及其产物的促淋巴细胞增殖活性[J].畜牧兽医学报,2010,41(6):741-745.
[18]FLEISCHER S,RIES S,SHEN P,et al.Anti-interleukin-6signalling therapy rebalances the disrupted cytokine production of B cells from patients with active rheumatoid arthritis[J].Eur J Immunol,2018,48(1):194-203.
[19]CHASSET F,ARNAUD L.Targeting interferons and their pathways in systemic lupus erythematosus[J].Autoimmun Rev,2018,17(1):44-52.
[20]YAO Y,WANG J B,XIN M M,et al.Balance between inflammatory and regulatory cytokines in systemic lupus erythematosus[J].Genet Mole Res,2016,15(2):1-8.
[21]徐毅芳,寿张飞.调节性T细胞在移植耐受中的作用[J].国际移植与血液净化杂志,2008,6(2):40-43.
[22]葛宽雪,高伟华,向晓星.辅助性T淋巴细胞17和调节性T淋巴细胞及其平衡改变与自身免疫性肝炎的关系[J].临床肝胆病杂志,2018,34(7):1573-1576.