丹参-苦参对免疫抑制小鼠的免疫调节功能探讨
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摘要
目的:通过使用环磷酰胺所致免疫抑制小鼠模型探讨丹参-苦参提取物对免疫缺陷小鼠的免疫调节作用的机制。方法:采用环磷酰胺造免疫抑制动物模型,空白组,模型组,丹参-苦参提取物低、中、高剂量组(25,50,100 mg·kg~(-1)),采用小鼠脏器指数评价脏器功能,采用小鼠碳粒廓清模型评价小鼠吞噬细胞功能,采用luminex法检测血清中相关细胞因子水平,流式检测辅助性T细胞数量。体外培养小鼠单核巨噬细胞RAW264. 7,给予丹参-苦参提取物后,采用噻唑蓝(MTT)比色法检测丹参-苦参提取物对RAW264. 7细胞增殖的影响,采用实时荧光定量聚合酶链式反应(Real-time PCR)对其增殖的影响及其可能机制进行探讨。结果:与空白组比较,模型组显著抑制肝指数(P <0. 01)和脾脏指数(P <0. 05),动物血清白细胞介素-6(IL-6)的分泌明显降低(P <0. 05),吞噬指数显著下降(P <0. 01),同时50 mg·L~(-1)环磷酰胺显著抑制RAW264. 7细胞的增殖(P <0. 01);与模型组比较,丹参-苦参提取物中、高剂量均可以显著提高肝脏指数(P <0. 01),并显著促进趋化因子C-X-C配体1(CXCL1)的分泌(P <0. 01),丹参-苦参提取物高剂量可以显著促进IL-6的分泌(P <0. 01),丹参-苦参提取物中、高剂量能够明显提高小鼠腋下以及颈部淋巴结中辅助性T细胞的数量(P <0. 05,P <0. 01),显著性升高吞噬指数的趋势(P <0. 01),显著性促进RAW264. 7细胞的增殖(P <0. 01),且促进IL-6信号传导与转录激活因子(STAT3)调控下游B淋巴细胞瘤-2(Bcl-2),Bcl-2样蛋白1(Bcl-2L1)以及细胞周期蛋白D1(CCND1) mRNA的表达,起抗凋亡作用。结论:丹参-苦参提取物可对抗环磷酰胺所导致的免疫抑制,增强动物免疫功能。
Objective: To investigate the mechanism of immunomodulatory effect of extracts from Salviae Miltiorrhizae Radix et Rhizoma and Sophorae Flavescentis Radix on immunodeficiency mice by immunosuppressive mouse model induced by cyclophosphamide. Method: An immunosuppressive animal model was established by cyclophosphamide. The blank group,the model group,the low,medium and high dose group of Salvia miltiorrhiza and Sophora extract groups( 25,50,100 mg·kg~(-1)),using mouse organs organ evaluation index using a mouse model to evaluate the carbon clearance phagocytic cell function using luminex to detect levels of relevant cytokines in serum and using flow to detect the number of helper T cells. Mouse mononuclear macrophage leukemia cells( RAW264. 7) were cultured in vitro,and the effect of Salvia miltiorrhiza extracts on the proliferation of RAW264. 7 cells were detected by methylthiazolyldiphenyl-tetrazolium bromide( MTT) assay. The proliferation was induced by Real-time PCR. The impact and its possible mechanisms are explored. Result: Compared with blank group,cyclophosphamide significantly inhibited liver index( P < 0. 01) and spleen index( P < 0. 05),and the secretion of serum interleukin-6( IL-6) was significantly decreased( P < 0. 05),the phagocytic index decreased significantly( P < 0. 01), while 50 mg·L~(-1) cyclophosphamide significantly inhibited the proliferation of mononuclear macrophage RAW264. 7( P < 0. 01). Compared with model group,high dose of extracts from Salviae Miltiorrhizae Radix et Rhizoma and Sophorae Flavescentis Radix can significantly increase the liver index( P < 0. 01). The high dose of extracts from Salviae Miltiorrhizae Radix et Rhizoma and Sophorae Flavescentis Radix can significantly promote the chemokine CXC motif ligand 1( CXCL1,P < 0. 01),high dose can significantly promote the secretion of IL-6( P < 0. 01). The high dose of extracts from Salviae Miltiorrhizae Radix et Rhizoma and Sophorae Flavescentis Radix can significantly improve the auxiliary in the axillary and cervical lymph nodes of mice. The number of T cells( P < 0. 05,P < 0. 01),the high dose of extracts from Salviae Miltiorrhizae Radix et Rhizoma and Sophorae Flavescentis Radix could significantly increase the phagocytic index( P < 0. 01). Medium and high saliency promoted the proliferation of RAW264. 7 cells( P < 0. 01),and promoted IL-6 signaling and activator of transcription 3( STAT3) regulation of the downstream B lymphoma-2 gene( Bcl-2),Bcl-2 like protein 1( Bcl-2 L1) and the cyclin D1 gene( CCND1) plays an anti-apoptotic role. Conclusion: The extracts from Salviae Miltiorrhizae Radix et Rhizoma and Sophorae Flavescentis Radix can counteract the immunosuppression caused by cyclophosphamide and enhance the immune function of animals.
Objective: To investigate the mechanism of immunomodulatory effect of extracts from Salviae Miltiorrhizae Radix et Rhizoma and Sophorae Flavescentis Radix on immunodeficiency mice by immunosuppressive mouse model induced by cyclophosphamide. Method: An immunosuppressive animal model was established by cyclophosphamide. The blank group,the model group,the low,medium and high dose group of Salvia miltiorrhiza and Sophora extract groups( 25,50,100 mg·kg~(-1)),using mouse organs organ evaluation index using a mouse model to evaluate the carbon clearance phagocytic cell function using luminex to detect levels of relevant cytokines in serum and using flow to detect the number of helper T cells. Mouse mononuclear macrophage leukemia cells( RAW264. 7) were cultured in vitro,and the effect of Salvia miltiorrhiza extracts on the proliferation of RAW264. 7 cells were detected by methylthiazolyldiphenyl-tetrazolium bromide( MTT) assay. The proliferation was induced by Real-time PCR. The impact and its possible mechanisms are explored. Result: Compared with blank group,cyclophosphamide significantly inhibited liver index( P < 0. 01) and spleen index( P < 0. 05),and the secretion of serum interleukin-6( IL-6) was significantly decreased( P < 0. 05),the phagocytic index decreased significantly( P < 0. 01), while 50 mg·L~(-1) cyclophosphamide significantly inhibited the proliferation of mononuclear macrophage RAW264. 7( P < 0. 01). Compared with model group,high dose of extracts from Salviae Miltiorrhizae Radix et Rhizoma and Sophorae Flavescentis Radix can significantly increase the liver index( P < 0. 01). The high dose of extracts from Salviae Miltiorrhizae Radix et Rhizoma and Sophorae Flavescentis Radix can significantly promote the chemokine CXC motif ligand 1( CXCL1,P < 0. 01),high dose can significantly promote the secretion of IL-6( P < 0. 01). The high dose of extracts from Salviae Miltiorrhizae Radix et Rhizoma and Sophorae Flavescentis Radix can significantly improve the auxiliary in the axillary and cervical lymph nodes of mice. The number of T cells( P < 0. 05,P < 0. 01),the high dose of extracts from Salviae Miltiorrhizae Radix et Rhizoma and Sophorae Flavescentis Radix could significantly increase the phagocytic index( P < 0. 01). Medium and high saliency promoted the proliferation of RAW264. 7 cells( P < 0. 01),and promoted IL-6 signaling and activator of transcription 3( STAT3) regulation of the downstream B lymphoma-2 gene( Bcl-2),Bcl-2 like protein 1( Bcl-2 L1) and the cyclin D1 gene( CCND1) plays an anti-apoptotic role. Conclusion: The extracts from Salviae Miltiorrhizae Radix et Rhizoma and Sophorae Flavescentis Radix can counteract the immunosuppression caused by cyclophosphamide and enhance the immune function of animals.
引文
[1]李金玉,李金耀,张富春.中药抗肿瘤免疫调节机制研究进展[J].时珍国医国药,2016,27(7):1705-1707.
[2]周悦芳,范培红.中药免疫调节作用研究进展[J].时珍国医国药,2017,28(1):204-207.
[3]蔡琳,彭鹏,郭甜.丹参药理作用及临床研究进展[J].山东化工,2016,45(17):51-52.
[4]王悦,姜雪,丁菲,等.中药苦参药理作用及应用研究进展[J].山东化工,2017,46(15):66-67,69.
[5]王和木.丹参酮联合苦参素治疗慢性乙肝的临床疗效分析[J].医学理论与实践,2015,28(9):1207-1209.
[6]郑洋,赵铁建.苦参素联合复方丹参治疗慢性乙型肝炎临床疗效的Meta分析[J].中国中西医结合消化杂志,2018,26(5):445-449.
[7]黄俊婷.苦参素注射液对晚期胃癌TP方案化疗的免疫调节作用[J].中国卫生产业,2014,11(8):137-138.
[8]张湘东,许定舟,李金华,等.丹参多糖的免疫调节活性研究[J].中药材,2012,35(6):949-952.
[9]张雯,宋俊科,王海港,等.破壁灵芝孢子粉对环磷酰胺致小鼠免疫抑制的调节作用[J].食品与药品,2018,20(6):408-413.
[10]蔡帆,张彦,臧林泉.白扁豆多糖对免疫抑制小鼠的免疫调节作用[J].免疫学杂志,2018,34(5):407-411.
[11]陈瑞明,张小丽,李芳,等.祛风湿类中药免疫作用的研究——对环磷酰胺致免疫功能低下小鼠免疫功能的影响[J].中华中医药学刊,2015,33(5):1171-1175.
[12]许良,李瑞瑞,李心雨,等.大蒜化学成分(组)对免疫抑制小鼠免疫功能的调节作用[J].西北药学杂志,2018,33(6):762-765.
[13] Ohkawara T,Mitsuyama K,Takeda H,et al. Lack of macrophage migration inhibitory factor suppresses innate immune response in murine dextran sulfate sodiuminduced colitis[J]. Scand J Gastroenterol,2008,43(12):1497-1504.
[14]张乔,姜苏洋,吕航,等.参苓水煎液对免疫抑制小鼠免疫调节作用的研究[J].中医药信息,2018,35(6):8-11.
[15] Pozios I,Knosel T,ZHAO Y,et al. Expression of phosphorylated estrogen receptor beta is an independent negative prognostic factor for pancreatic ductal adenocarcinoma[J]. J Cancer Res Clin Oncol,2018,144(10):1887-1897.
[16] BAI Y,JIANG Y,LIU T,et al. Xinjiang herbal tea exerts immunomodulatory activity via TLR2/4-mediated MAPK signaling pathways in RAW264. 7 cells and prevents cyclophosphamide-induced immunosuppression in mice[J]. J Ethnopharmacol, 2019, doi:10. 1016/j.jep. 2018. 09. 032.
[17]刚宏林,于雅楠,马英南,等.玻璃苣水提物对荷瘤小鼠免疫调节作用的实验研究[J].中医药信息,2016,33(1):25-28.
[18]马红芳,苏国新,朱小丽,等.大株红景天注射液对乳腺癌化疗患者免疫功能调节的临床观察[J].中国实验方剂学杂志,2016,22(21):172-176.
[19]杨继乐,张莉,王莉.单核-巨噬细胞的分化和功能研究进展[J].细胞与分子免疫学杂志,2014,30(11):1213-1216.
[20]孟祥云,郭树明,杨丽霞.中药植物多糖对2型糖尿病胰岛素抵抗的作用机制研究进展[J].中国实验方剂学杂志,2017,23(8):220-225.
[21]廖海锋,邓向亮,罗霞,等.羧甲基茯苓多糖对巨噬细胞极化的影响[J].中国实验方剂学杂志,2016,22(13):122-126.
[2]周悦芳,范培红.中药免疫调节作用研究进展[J].时珍国医国药,2017,28(1):204-207.
[3]蔡琳,彭鹏,郭甜.丹参药理作用及临床研究进展[J].山东化工,2016,45(17):51-52.
[4]王悦,姜雪,丁菲,等.中药苦参药理作用及应用研究进展[J].山东化工,2017,46(15):66-67,69.
[5]王和木.丹参酮联合苦参素治疗慢性乙肝的临床疗效分析[J].医学理论与实践,2015,28(9):1207-1209.
[6]郑洋,赵铁建.苦参素联合复方丹参治疗慢性乙型肝炎临床疗效的Meta分析[J].中国中西医结合消化杂志,2018,26(5):445-449.
[7]黄俊婷.苦参素注射液对晚期胃癌TP方案化疗的免疫调节作用[J].中国卫生产业,2014,11(8):137-138.
[8]张湘东,许定舟,李金华,等.丹参多糖的免疫调节活性研究[J].中药材,2012,35(6):949-952.
[9]张雯,宋俊科,王海港,等.破壁灵芝孢子粉对环磷酰胺致小鼠免疫抑制的调节作用[J].食品与药品,2018,20(6):408-413.
[10]蔡帆,张彦,臧林泉.白扁豆多糖对免疫抑制小鼠的免疫调节作用[J].免疫学杂志,2018,34(5):407-411.
[11]陈瑞明,张小丽,李芳,等.祛风湿类中药免疫作用的研究——对环磷酰胺致免疫功能低下小鼠免疫功能的影响[J].中华中医药学刊,2015,33(5):1171-1175.
[12]许良,李瑞瑞,李心雨,等.大蒜化学成分(组)对免疫抑制小鼠免疫功能的调节作用[J].西北药学杂志,2018,33(6):762-765.
[13] Ohkawara T,Mitsuyama K,Takeda H,et al. Lack of macrophage migration inhibitory factor suppresses innate immune response in murine dextran sulfate sodiuminduced colitis[J]. Scand J Gastroenterol,2008,43(12):1497-1504.
[14]张乔,姜苏洋,吕航,等.参苓水煎液对免疫抑制小鼠免疫调节作用的研究[J].中医药信息,2018,35(6):8-11.
[15] Pozios I,Knosel T,ZHAO Y,et al. Expression of phosphorylated estrogen receptor beta is an independent negative prognostic factor for pancreatic ductal adenocarcinoma[J]. J Cancer Res Clin Oncol,2018,144(10):1887-1897.
[16] BAI Y,JIANG Y,LIU T,et al. Xinjiang herbal tea exerts immunomodulatory activity via TLR2/4-mediated MAPK signaling pathways in RAW264. 7 cells and prevents cyclophosphamide-induced immunosuppression in mice[J]. J Ethnopharmacol, 2019, doi:10. 1016/j.jep. 2018. 09. 032.
[17]刚宏林,于雅楠,马英南,等.玻璃苣水提物对荷瘤小鼠免疫调节作用的实验研究[J].中医药信息,2016,33(1):25-28.
[18]马红芳,苏国新,朱小丽,等.大株红景天注射液对乳腺癌化疗患者免疫功能调节的临床观察[J].中国实验方剂学杂志,2016,22(21):172-176.
[19]杨继乐,张莉,王莉.单核-巨噬细胞的分化和功能研究进展[J].细胞与分子免疫学杂志,2014,30(11):1213-1216.
[20]孟祥云,郭树明,杨丽霞.中药植物多糖对2型糖尿病胰岛素抵抗的作用机制研究进展[J].中国实验方剂学杂志,2017,23(8):220-225.
[21]廖海锋,邓向亮,罗霞,等.羧甲基茯苓多糖对巨噬细胞极化的影响[J].中国实验方剂学杂志,2016,22(13):122-126.