茵陈蒿汤保肝作用的机理Ⅲ.对小鼠腹腔巨噬细胞释放NO的影响

目的：探讨茵陈蒿汤醇提物保肝作用与一氧化氮（ＮＯ）的关系。方法：ＮＯ以Ｇｅｉｅｓ试剂测定。结果：茵陈蒿汤醇提物０２～２０μｇ直接加入小鼠腹腔巨噬细胞培养液可抑制ＮＯ产生,含茵陈蒿汤醇提物小鼠血清１００,１５０,２００ｍｉｎ对ＮＯ有促进作用,效应高峰在１００ｍｉｎ。结论：调节巨噬细胞释放ＮＯ是茵陈蒿汤醇提物保肝途径之一。     

小柴胡汤对小鼠腹腔巨噬细胞释放 NO 的影响

为了了解小柴胡汤抗肝损害机理，采用Ｇｒｉｅｓｓ度剂检测小柴胡汤醇提物及灌服小柴胡汤醇提物小鼠血清对小鼠巨噬细胞释放ＮＯ的影响。结果显示：小柴胡汤醇提物直接加入小鼠腹腔巨噬细胞培养液中不影响ＮＯ的产生；而加入１２０ｍｉｎ，１８０ｍｉｎ，２４０ｍｉｎ采集的含小柴胡汤醇提物小鼠血清能促进ＮＯ的产生，结果显示：促进巨噬细胞释放ＮＯ可能是小柴胡汤抗肝损害的机理之一。     

中药复方作用机理研究举例

中药尤其是复方的作用机理研究迄今为止都比较困难,方法亦不多,本文应用细胞培养和血清药理方法对菌开蒿汤的保肝机理进行了研究,以探讨该方法在中药复方作用机理研究中应用的可行性。茵陈蒿汤出自《伤寒论》,是治疗湿热黄疸的要方,临床用于多种黄疸症及传染性肝炎、急性病毒性肝炎、重型肝炎的治疗。本文研究了茵陈蒿汤醇提物对小鼠巨噬细胞释放TNF的影响,探讨其保肝作用与TNF的关系。     

茵陈蒿汤醇提物对机体抗氧化能力影响

为探讨茵陈蒿汤醇提物的保肝作用与机体抗氧化能力的关系,实验观察茵陈蒿汤醇提物在Ｄ─氨基半乳糖＋脂多糖（Ｄ—ＧａｌＮ＋ＬＰＳ）和卡介苗＋脂多糖（ＢＣＧ＋ＬＰＳ）模型中对ＡＬＴ、ＧＳＴ、ＬＰＯ、ＳＯＤ及ＧＳＨ的影响。结果：茵陈蒿汤醇提物ｉｇ．６ｇ／ｋｇ和９ｇ／ｋｇ明显降低Ｄ—ＣａｌＮ＋ＬＰＳ肝损伤小鼠血清谷丙转氨酶（ＡＬＴ）活性,降低谷胱甘肽─Ｓ─转移酶（ＧＳＴ）水平,减少肝匀浆中脂质过氧化物（ＬＰＯ）含量,对超氧化物歧化酶（ＳＯＤ）活性无明显影响;４ｇ／ｋｇ,６ｇ／ｋｓ和９ｇ／ｋｓ显著降低ＢＣＧ＋ＬＰＳ免疫性肝损伤小鼠血清ＡＬＴ水平,６ｇ／ｋｇ和９ｇ／ｋｇ增加肝匀浆还原型谷胱甘肽（ＧＳＨ）含量。表明茵陈蒿汤醇提物经口给药能明显提高小鼠抗自由基损伤的能力,为其保肝作用的机理之一。     

茵陈蒿汤提取物保肝作用的血清药理学研究

茵陈蒿汤出自《伤寒论》,由茵陈蒿、栀子、大黄组成,能清热利湿,解毒退黄,主要用于治疗湿热黄疸症。现代研究表明,它具有利胆、保胆、解热抗炎镇痛、抗菌抗病毒的作用。利胆保肝作用是本方临床治疗多种黄疸及传染性肝炎、急性肝炎、急性病毒性肝炎、重型肝炎的主要药理基础。本实验室既往研究已观察到茵陈蒿汤醇提物的保肝作用优于其水提物。本文通过CCl4、D-GalN LPS和BCG LPS诱导小鼠肝损伤观察茵陈蒿汤醇提物的氯仿、乙酸乙酯萃取部位及剩余乙醇提取物降低ALT活性的作用,试图分析茵陈蒿汤保肝作用的效应物质基础。1实验材料1.1动…     

茵陈蒿汤醇提物保肝作用分析

目的:观察茵陈蒿汤醇提物的氯仿、乙酸乙酯萃取部位对D-GalN诱导的大鼠肝细胞损伤的影响。方法:用氯仿、乙酸乙酯分别萃取茵陈蒿汤醇提物得到氯仿部位、乙酸乙酯部位及剩余乙醇提取物,观察其对D-GalN诱导的大鼠肝细胞损伤的保护作用。结果:氯仿部位及乙醇提取物有降低ALT活性的作用,且氯仿部位优于乙醇提取物,而乙酸乙酯部位无降低ALT活性的作用。结论:茵陈蒿汤保肝作用的效应物质可能富集在氯仿萃取部位。     

茵陈蒿汤保肝作用有效成分分析

目的：分析茵陈蒿汤保肝作用有效成分。方法：用75%乙醇分别提取茵陈蒿，栀子，大黄及茵陈蒿汤。观察其对D-GalN诱导的大鼠肝细胞损伤的保护作用。结果：茵陈蒿醇提物对D-GalN诱导的大鼠肝细胞损伤ALT升高的降低作用优于茵陈蒿醇提物，栀子醇提物及大黄醇提物，结论：茵陈蒿汤保肝作用可能是茵陈蒿，栀子，大黄共同作用的结果。     

大枣多糖体外对小鼠腹腔巨噬细胞功能的影响

目的：探讨大枣多糖（ＪＤＰＮ）体外对小鼠腹腔巨噬细胞细胞毒活性和分泌ＴＮＦα、ＩＬ１、ＮＯ等功能的影响。方法：ＭＴＴ法测细胞增殖活性;胸腺细胞增殖法测ＩＬ１;Ｌ９２９细胞株细胞毒法测ＴＮＦα;Ｇｒｅｅｓ试剂测ＮＯ。结果：ＪＤＰＮ能显著增强巨噬细胞（ＭΦ）对Ｌ９２９细胞株的细胞毒活性和ＴＮＦα、ＩＬ１、ＮＯ的分泌功能。ＪＤＰＮ最佳诱导浓度对促进ＭΦ分泌ＩＬ１为５０μｇ／ｍｌ,促ＴＮＦα分泌为１００μｇ／ｍｌ。ＪＤＰＮ不能协同ＬＰＳ促进ＩＬ１和ＴＮＦα的分泌。１００μｇ／ｍｌＪＤＰＮ诱导ＭΦ分泌ＮＯ能力最强。结论：大枣多糖体外可增强小鼠腹腔巨噬细胞的细胞毒作用,诱导ＴＮＦα、ＩＬ１、ＮＯ的产生。     

一点红提取物药理作用的实验研究

目的:观察一点红提取物的药理作用。方法:采用小鼠腹腔毛细血管通透性实验,测定伊文思蓝光密度值;采用小鼠扭体法,测定醋酸所致小鼠扭体次数;用CCl4肝损伤模型,测定小鼠血清中转氨酶活性及蛋白含量;采用小鼠腹腔巨噬细胞吞噬功能试验,腹腔巨噬细胞吞噬百分率和吞噬指数;并做小鼠急性毒性试验,测定小鼠的LD50。结果:一点红水提物大剂量组和醇提物大小剂量组均能明显抑制小鼠腹腔毛细血管通透性;一点红水提物小剂量组、一点红醇提物大小剂量组均能明显减少醋酸所致小鼠扭体次数;一点红水提物和醇提物大剂量组对CCl4所致急性肝损伤小鼠血清中ALT、AST活性升高有抑制作用;一点红水提物和醇提物大、小剂量组和左旋咪唑组均能明显增加免疫功能低下小鼠腹腔巨噬细胞吞噬百分率和吞噬指数;一点红水提物的LD50为(49.22±0.027)g生药/kg,一点红醇提取物的LD50为(63.15±0.026)g生药/kg。结论:一点红水提物和醇提物具有一定的抗炎、镇痛、保肝、免疫增强作用。     

小红参含药血清对链球菌抗原刺激小鼠的免疫药理学研究

目的利用血清药理学实验方法,评价小红参含药血清对链球菌抗原刺激小鼠的免疫药理学效应,初步探讨小红参治疗银屑病的作用机制。方法利用β溶血型链球菌的超声破碎抗原为刺激物,观察小红参含药血清对链球菌致敏小鼠脾脏淋巴细胞的增殖活性,以及对小鼠腹腔巨噬细胞分泌一氧化氮(NO)的影响。结果醇提物各剂量组以及水提物高剂量组与空白血清组比较有显著性差异,均对链球菌抗原致敏小鼠的T淋巴细胞增殖具有抑制作用。醇提物高、中剂量组以及水提物高剂量组与空白血清组比较有显著性差异,均能抑制链球菌抗原致敏小鼠的腹腔巨噬细胞NO的分泌水平。结论小红参含药血清能够明显抑制T淋巴细胞对链球菌抗原的增殖作用;小红参含药血清可以抑制链球菌抗原诱导的小鼠腹腔巨噬细胞NO的分泌。     

Acanthopanax senticosus suppresses reactive oxygen species production by mouse peritoneal macrophages in vitro and in vivo

Excess production of reactive oxygen species by macrophages has been implicated in many inflammatory diseases. The present study investigated the inhibitory effect of the stem bark extract of Acanthopanax senticosus on the production of superoxide anion and hydrogen peroxide in mouse peritoneal macrophages in vitro and in vivo. Exposure of mouse peritoneal macrophages to A. senticosus extract significantly suppressed superoxide anion production induced by zymosan in a dose-dependent manner. Similarly, exposure of mouse peritoneal macrophages to A. senticosus extract significantly inhibited hydrogen peroxide production induced by phorbol 12-myristate 13-acetate (PMA) in a dose-dependent manner. Intraperitoneal administration of A. senticosus extract to KM mice reduced the ex vivo production of zymosan induced-superoxide anion and PMA-induced hydrogen peroxide by their peritoneal macrophages. Exposure to A. senticosus extract did not affect the cell viability or systemic toxicity. A. senticosus inhibited reactive oxygen species production by mouse peritoneal macrophages in vitro and in vivo and may be partly responsible for the antiinflammatory function.     

玛咖-人参水煎液对小鼠免疫功能的影响

目的:探讨玛咖-人参水煎液对小鼠免疫功能的影响。方法:通过碳廓清试验、小鼠腹腔巨噬细胞吞噬鸡红细胞试验、绵羊红细胞诱导的小鼠迟发型变态反应试验、ConA诱导的小鼠淋巴细胞转化试验(MTT法)、Jerne改良玻片法测定抗体生成细胞数试验、小鼠血清溶血素的测定,研究玛咖-人参水煎液对小鼠免疫功能的影响。结果:与空白组比较,玛咖-人参水煎液低、中、高剂量组均可明显提高小鼠吞噬指数(P0.05或P0.01),玛咖-人参水煎液低、中、高剂量组均可明显提高小鼠巨噬细胞吞噬鸡红细胞吞噬率(P0.05或P0.01)和吞噬指数(P0.01),玛咖-人参水煎液中、高剂量组可明显提高小鼠足趾肿胀度(P0.05),玛咖-人参水煎液高剂量组可明显提高小鼠溶血空斑数(P0.01)。结论:玛咖-人参水煎液能提高小鼠单核-巨噬细胞功能、细胞免疫功能和体液免疫功能。     

Acanthopanax senticosus inhibits nitric oxide production in murine macrophages in vitro and in vivo

Excess nitric oxide (NO) production has been implicated in inflammatory diseases. The present study investigated the inhibitory effect of the stem bark extract of Acanthopanax senticosus (A. senticosus) on NO production in murine macrophages in vitro and in vivo. In vitro exposure of RAW264.7 cells to 1, 10, 50, 100, 250, 500 and 1000 microg/mL of A. senticosus extract significantly suppressed NO production induced by lipopolysaccharide (LPS) and interferon gamma (IFN-gamma) in a dose-dependent manner. In vitro exposure of mouse resident peritoneal macrophages to 1, 10, 100 and 1000 microg/mL of A. senticosus extract significantly suppressed NO production induced by LPS and IFN-gamma in a dose-dependent manner. In vivo administration of A. senticosus extract (50, 100 and 200 mg/kg) to KM mice dose-dependently inhibited LPS and IFN-gamma induced production of NO in isolated mouse peritoneal macrophages ex vivo. Exposure to A. senticosus extract had no effect on cell viability and systemic toxicity. The results demonstrated that the stem bark extract of A. senticosus extract inhibits NO production in murine macrophages in vitro and in vivo.     

商陆提取物醋制前后毒性作用的比较研究

目的:提取分离获得商陆的毒性成分,并比较其醋制前后毒性的变化.方法:采用黏膜刺激性反应、小鼠腹腔炎症模型及离体巨噬细胞释放NO模型,比较商陆毒性成分醋制前后的致炎毒性变化.结果:商陆毒性成分具有显著的致炎毒性,可导致家兔眼结膜水肿、小鼠腹腔渗出液中PGE2含量升高以及巨噬细胞释放NO含量升高;经醋制后,对家兔眼结膜刺激性减弱,使小鼠腹腔渗出液中PGE2含量降低、巨噬细胞释放NO含量降低.结论:醋制法能够降低商陆毒性成分的毒性作用.     

两种哒嗪酮衍生物对小鼠腹腔巨噬细胞体外释放肿瘤坏死因子的影响

 目的研究两种哒嗪酮衍生物即：6-(a,a-二苯基乙酰哌嗪基苯基)-4,5-二氢-5-甲基-3-(2H)-哒嗪酮(简称DMDP)和6-(a-苯基乙酰哌嗪基苯基)-4,5-二氢-5-甲基-3-(2H)-哒嗪酮(简称PMDP)及对照药物维拉帕米(verapamil,Ver)对小鼠腹腔巨噬细胞体外释放肿瘤坏死因子(tumor necrosis factor,TNF)的影响。方法体外诱生TNF,以L929细胞为靶细胞的细胞毒结晶紫染色法进行TNF的生物定量测定。结果在10^-6～10^-4mol·L^-1浓度范围内,DMDP的抑制作用呈剂量依赖性;浓度达10^-4mol·L^-1时,可完全抑制TNF的释放。IC50<10-6mol·L^-1。PMDP的抑制作用更强：10^{-8～10-6mol.L-1时,该抑制作用呈剂量依赖性;10-6mol·L-1时,即可完全抑制TNF的释放。IC50<1O-8mol·L-1。结论DMDP,PMDP及Ver对小鼠腹腔巨噬细胞体外释放TNF具有显著的抑制作用。}     

石榴皮乙醇提取物对小鼠细胞免疫功能的影响

目的：探讨石榴皮乙醇提取物对小鼠细胞免疫功能的影响。方法：通过腹腔注射环磷酰胺建立免疫抑制动物模型,应用石榴皮乙醇提取物、石榴皮总黄酮提取物对免疫抑制的小鼠进行治疗,观察两种提取物对小鼠巨噬细胞吞噬功能、脾淋巴细胞转化功能的影响。结果：石榴皮乙醇提取物中剂量组和高剂量组及石榴皮总黄酮提取物组能增强小鼠腹腔巨噬细胞的吞噬功能,高剂量组能够增强小鼠脾淋巴细胞转化功能（P<0.01）。结论：石榴皮乙醇提取物对免疫抑制小鼠的细胞免疫功能具有一定的提高作用。     

Suppression of nitric oxide production in activated murine peritoneal macrophages in vitro and ex vivo by Scrophularia striata ethanolic extract

Ethnopharmacological relevance

Scrophularia striata (Scrophulariaceae), a traditional Iranian medicine, has been used for the treatment of allergy, rheumatics and chronic inflammatory disorders.

Aim of the study

In the present study, we investigated the in vitro and ex vivo suppressive effects of Scrophularia striata ethanolic extract on nitric oxide production in mouse peritoneal macrophages.

Materials and methods

Peritoneal macrophages were harvested by lavaging with ice cold phosphate buffer saline. Macrophages obtained from mice not treated were cultured with 10 μg/mL lipopolysaccaride (LPS), 20 U/mL interferon-γ (IFN-γ), and various concentrations of Scrophularia striata extract for the in vitro experiments and those obtained from mice treated with different doses of the extract for 7 days were cultured with 10 μg/mL LPS, 20 U/mL IFN-γ for the in vivo experiments. Nitrit levels were measured by using the diazotization method based on the Griess reaction, which is an indirect assay for NO production.

Results

In vitro exposure of mouse peritoneal macrophages with various concentrations of Scrophularia striata extract (10, 50 and 100 μg/mL) significantly suppressed NO production in a dose-dependent manner. In vivo administration of Scrophularia striata extract (50 and 100 mg/kg) to Balb/c mice inhibited LPS and IFN-γ induced production of NO in the isolated mouse peritoneal macrophages ex vivo in a dose-dependent manner. Exposure to Scrophularia striata extract had no effect on cell viability.

Conclusion

The results of the study demonstrated that the Scrophularia striata extract inhibit NO production in activated murine macrophages and we suggest that Scrophularia striata may be used in treating the inflammatory diseases.     

益气、活血、益气活血类中药复方对S180荷瘤小鼠免疫抑瘤作用的比较研究

目的：比较益气、活血、益气活血类中药复方对S180荷瘤小鼠免疫抑瘤作用。方法：本研究以S180荷瘤小鼠为实验对象，对补气的四君子汤、活血的桃红四物汤、益气活血的加味八珍汤对免疫功能及抑瘤作用的影响做对比研究。结果：荷瘤对照组的NK活性、IL-2分泌水平及腹腔巨噬细胞吞噬能力均明显低于正常对照组，经灌胃中药2周后，益气组、活血组、益气活血组的NK活性、IL-2分泌水平及腹腔巨噬细胞吞噬能力均接近或高于正常对照组。以加味八珍汤治疗组的效果最好。且加味八珍汤组的抑瘤率高达45．9％。结论：在肿瘤的治疗中，将益气活血类方药联合应用的作用强于单纯的益气组和活血组。