中医体质学基础实验方法和研究现状

基础实验研究是中医体质学的重要研究方法之一。生物化学、免疫遗传学、生物物理学、基因组学、代谢组学、蛋白组学等实验方法被应用于中医体质学研究。但目前体质基础研究仍存在报道数量较少,实验数据缺乏深入分析等不足之处。今后中医体质实验研究在思路上仍然应以整体性和系统性为指导,宏观与微观相结合,方法学上则应当充分借鉴表观遗传学、免疫组库等现代科学新的研究理念和实验技术。并且应加强基础研究的投入,以期取得突破性进展。     

冠脉康胶囊对大鼠血栓模型及小鼠急性脑缺血模型的影响

[目的]复制大鼠血栓模型及小鼠急性脑缺血模型,观察冠脉康胶囊对大鼠血栓模型及对小鼠急性脑缺血模型的影响。[方法]采用结扎大鼠左肾下方下腔静脉致血栓模型形成法,观察冠脉康胶囊抗血栓形成作用;采用小鼠断头缺血模型,观察冠脉康胶囊抗急性脑缺血作用。[结果]冠脉康胶囊能减轻血栓模型大鼠的血栓重量,增加小鼠张口呼吸次数,延长小鼠张口呼吸时间。[结论]冠脉康胶囊具有抗血栓形成、抗急性脑缺血作用。     

Granulocyte-macrophage colony stimulating factor inhibits class II major histocompatibility complex expression and antigen presentation by microglia

Granulocyte-macrophage colony stimulating factor (GM-CSF) modulates various functions of monocytes/ macrophages including antigen-presenting capacity. Recently it was found that astrocytes produce GM-CSF in the central nervous system (CNS) and that GM-CSF can induce proliferation and morphological changes of microglia. Here we show that GM-CSF can down regulate the interferon-γ-mediated induction of major histocompatibility complex (MHC) class II antigens in microglia, but not in astrocytes. GM-CSF pretreatment completely prevents myelin basic protein-specific T cell proliferation induced by microglia but not astrocytes. GM-CSF did not affect the cell surface expression on microglia of either MHC class I or cell adhesion molecules. The inhibition of microglial MHC class II expression and antigen-presenting function is specific for GM-CSF, as treatment with a different CSF (interleukin-3) did not modulate microglial phenotype or functional capacity. These data suggest that GM-CSF might be involved in the regulation of immune responses within the central nervous system.     

Augmentation of adoptively transferred experimental allergic encephalomyelitis by administration of a monoclonal antibody specific for LFA-1α

We investigated the effect of an anti-leukocyte function antigen 1 (LFA-1α) monoclonal antibody, M17/4.2, on murine relapsing experimental allergic encephalomyelitis (EAE). In vitro investigations demonstrated that M17/4.2 inhibited proliferation with concanavalin A or myelin basic protein. Control mice treated with phosphate buffered saline (PBS) developed a mild to moderate disease at 7–10 days followed by a long-term relapsing clinical course. With administration of M17/4.2, the time of disease onset was unchanged; however, the severity of the disease was greatly augmented, resulting in early mortality. The pathology correlated well with the clinical course. M17/4.2 mice showed more inflammation and demyelination than PBS or anti-CD4 treated mice. Therefore, this anti-LFA-1 specific monoclonal antibody augmented EAE.     

Effects of Danqidihuang Granules on glucolipid metabolism in insulin-resistant rats

Objective

To explore whether the insulin resistance (IR) model could be established through feeding Sprague-Dawley (SD) rats high-sugar and high-fat diets and to further observe the preventive and treatment effects of different doses of Danqidihuang Granules in rats.

Methods

Thirty-two SD rats were divided randomly into control group A (given regular feed), model group B (food high in sugar and fat), intervention group C (food high in sugar and fat as well as regular doses of Danqidihuang Granules), and intervention group D (food high in sugar and fat as well as double doses of Danqidihuang Granules). The interventions were for 8 weeks. Motion, change in color, body weight, and food intake, as well as plasma lipids (including low-density lipoprotein-cholesterol (LDL-C), high-density lipoprotein-cholesterol (HDL-C), total cholesterol (TC) and triglyceride (TG), fasting blood glucose (FBG), fasting insulin (FINs) levels, insulin sensitivity index (ISI), and insulin resistance index (HOMO-IR) were observed.

Results

At the end of the second week of the experiment, the appetite and activities of rats in groups B, C and D decreased significantly compared with group A. The fur of the rats in those three groups was curly. After the fourth week, the activities, food intake and color of rats in group B were worse than those in groups C and D, but there were no significant differences in weight (P>0.05). Compared with group A, LDL-C, TC, FBG and HOMO-IR in model group B were increased significantly (P<0.05), whereas the FINs and ISI increased obviously (P<0.05). The levels of LDL-C and TC in group D was decreased obviously compared with those in group C, and HOMO-IR in group D was less than that in group B (P<0.05).

Conclusions

Danqidihuang Granules helped to prevent and improved the insulin resistance of rats.     

二冬汤含药血清对肺癌细胞A549的作用研究

目的：研究二冬汤含药血清对肺癌细胞A549的抑制作用、最佳给药方案及诱导细胞凋亡。方法：采用MTT法,观察不同给药天数、不同剂量、不同采血时间、不同培养时间的二冬汤血清对肺癌细胞A549的抑制效应,及应用FITC／PI流式细胞术检测最佳给药方案含药血清对A549细胞的凋亡率。结果：二冬汤含药血清对人肺癌细胞A549具有抑制作用,中剂量组[7．99g／（kg·d）]给药5天、10天,末次给药2h的含药血清与细胞培养48h后的抑制作用最明显,抑制率分别为42．27％和39．67％,与空白血清组比较,差异均有非常显著性意义（P〈0．01）;但从给药天数以及与肺癌细胞培养时间上来看,与空白血清组比较,差异无显著性意义（P〉0．05）;FITC／PI双染法显示二冬汤含药血清对A549的凋亡总计为（28．45±0．68）％,与空白血清组（6．75±0．46）％比较,差异有非常显著性意义护〈0．01）。结论：二冬汤最佳给药方案为给药5天,末次给药2h后取血,此时制备的含药血清能诱导细胞发生凋亡。     

金铃子散镇痛作用的实验研究

目的:观察金铃子散水煎液的镇痛作用。方法:采用热板法及醋酸扭体法建立小鼠疼痛模型,观察金铃子散水煎液对热板法所致小鼠出现舔足时间(痛阈值)及醋酸扭体法所致小鼠扭体次数(15min内)的影响。结果:金铃子散水煎液高剂量组可使热板法小鼠在给药后30min、60min及90min痛阈值明显延长,醋酸所致小鼠扭体次数明显减少(P0.05,P0.01)。结论:金铃子散水煎液具有镇痛作用。     

益气化瘀冲剂防治大鼠早期心力衰竭的作用机制

目的探讨益气化瘀冲剂防治大鼠早期心力衰竭的作用机制。方法从70只SD大鼠中随机取10只为空白对照组,余60只均采用腹主动脉缩窄术制备早期心力衰竭大鼠模型。造模大鼠6周内死亡20只,存活40只。将存活的40只大鼠随机分为4组,即模型对照组、贝那普利对照组及益气化瘀冲剂高、低剂量组,各10只。造模后6周开始给药,时间8周。空白对照组、模型对照组均予蒸馏水1 mL/(100 g.d)灌胃;贝那普利对照组予贝那普利混悬液0.36 mg/(100 g.d)灌胃;益气化瘀冲剂高、低剂量组分别予益气化瘀冲剂4.0、1.0 g/(100 g.d)灌胃。检测各组大鼠血流动力学指标及左心室质量指数。结果模型对照组大鼠血流动力学指标与空白对照组比较差异均有统计学意义(P<0.05,P<0.01),提示造模成功。益气化瘀冲剂高剂量组平均动脉压(MBP)、左室内压上升最大速率及左室内压下降最大速率与贝那普利对照组比较差异均有统计学意义(P<0.05)。益气化瘀冲剂高、低剂量组左室内压上升、下降最大速率比较差异有统计学意义(P<0.05)。贝那普利对照组及益气化瘀冲剂高、低剂量组左心室质量、左心室质量指数与模型对照组比较差异均有统计学意义(P<0.05,P<0.01)。结论益气化瘀冲剂能抑制大鼠心室重构的发展,在改善心力衰竭大鼠血流动力学方面,高剂量益气化瘀冲剂优于贝那普利;在改善左心室质量、质量指数方面,益气化瘀冲剂与贝那普利作用相似,且高剂量益气化瘀冲剂作用优于低剂量益气化瘀冲剂。     

参灵方防治佐剂性关节炎大鼠药源性胃黏膜损伤及对前列腺素E2含量的影响

目的探讨参灵方防治佐剂性关节炎(AA)大鼠药源性胃黏膜损伤及对前列腺素(PG)E2含量影响。方法从70只Wistar大鼠中随机取出10只作为空白对照组,其余60只均制作AA大鼠模型,将出现二次反应的大鼠随机取出40只,并随机分为4组,即模型对照组、雷尼替丁对照组及参灵方小、大剂量组,各10只。雷尼替丁对照组予盐酸雷尼替丁胶囊30 mg/(kg.d)灌胃,参灵方小、大剂量组分别予参灵方3.5、7.0 g/(kg.d)灌胃,空白对照组、模型对照组分别予等容积0.9%氯化钠注射液灌胃。灌胃时间均为28 d,于末次灌胃结束后模型对照组与各给药组均采用腹部皮下注射吲哚美辛进一步诱导药源性胃黏膜损伤的复合模型,计算胃黏膜损伤指数、损伤抑制率,测定胃黏膜组织及血清中PGE2含量。结果雷尼替丁对照组及参灵方大、小剂量组胃黏膜损伤指数均较模型对照组减少(P<0.01);参灵方大、小剂量组胃黏膜损伤指数比较差异有统计学意义(P<0.05)。空白对照组大鼠胃黏膜色泽红润,未见损伤性改变;模型对照组大鼠胃黏膜表面可见点线状出血,少数呈斑片状;各用药组均仅见针尖样出血及点状糜烂,其中以雷尼替丁组及参灵方小剂量组较好,参灵方大剂量组次之。与空白对照组比较,模型对照组胃黏膜组织及血清中PGE2含量均降低(P<0.05);与模型对照组比较,雷尼替丁对照组及参灵方大、小剂量组胃黏膜组织及血清中PGE2含量均升高(P<0.05)。结论参灵方对AA大鼠药源性胃黏膜损伤有保护作用。     

病理生理学实验教学改革探索

第三军医大学病理生理学教研室对传统病理生理学实验教学进行了改革，在实验教学方法改进、实验内容优化、实验教材改革与更新、实验设计课程开设以及综合考核评价体系建立等方面进行了有益的探索，取得良好的效果。