基于《内经》肝“生血气”理论探讨调肝生血法对复合病因贫血小鼠的补血效应

发布时间：2017-12-27 22:22

本文关键词：基于《内经》肝“生血气”理论探讨调肝生血法对复合病因贫血小鼠的补血效应　出处：《成都中医药大学》2016年博士论文　论文类型：学位论文

【摘要】：目的：应用调肝生血法,观察小柴胡汤、当归补血汤、小柴胡合当归补血汤、自拟柴胡生血方对失血加饥饿复合病因造模法制备的贫血小鼠的补血效应,探讨《黄帝内经》“肝生血气”理论,为临床治疗贫血提供新的思路和方法,为“肝生血气”理论提供实验研究依据。方法：1.对《中华医典》中1000多种中医古代医籍有关“肝生血”理论的疏理和分析,总结归纳“肝生血”理论。2.选取SPF级小鼠雌雄各半分笼饲养,将小鼠随机分为空白组、模型组、小柴胡汤组、当归补血汤组、小柴胡合当归补血汤组、柴胡生血方组,用复合病因造模法制备贫血小鼠模型,周期14天。各治疗组于第15天开始,每日进行中药汤剂灌胃,7天为1疗程,共2个疗程。①观察血虚小鼠体重、体态改变；②分别于灌药1周、2周后,每组随机选取雌雄各4只小鼠采用摘眼球取血的方法取血,进行外周血血常规检测、采用Elisa法检测小鼠血清细胞因子SCF、EPO、 IL-3、TGF-β1、TNF-α含量：③处死动物摘取小鼠肝脏、心脏、脑制作病理组织切片,观察小鼠脑、心、肝组织缺血的病理改变,并评分统计。结果：1.体重变化：造模前、后,实验小鼠体重变化有显著性差异(P0.01)；灌药1周后,空白组和模型组比较,灌药组和模型组比较,体重变化无显著性差异(P0.05)；灌药第2周后,与空白组比较,模型组体重小于空白组,有显著性差异(P0.05),与模型组比较,当归补血汤组、小柴胡合当归补血汤组、柴胡生血方组小鼠体重增重,差异有显著性意义(P0.05)。其余各组间的比较,空白组、模型组、当归补血汤组、小柴胡汤组、小柴胡合当归补血汤组5组小鼠的体重均小于柴胡生血方组小鼠体重,且差异有显著性(P0.05)。2.血常规结果：与模型组比较,各用药组对外周血象有不同程度的提高,灌药1周后,小柴胡合当归补血汤组、小柴胡汤组小鼠血常规RBC、HGB含量显著高于模型组(P0.05),灌药2周后,小柴胡合当归补血汤组RBC显著高于模型组(P0.05),柴胡生血方组HGB、WBC含量显著高于模型组(P0.05)。3.血清中细胞因子指标检测结果：(1)灌药1周后与模型组比较,小柴胡汤组、小柴胡合当归补血汤组、柴胡生血方组EPO含量显著增高(P0.05)；灌药2周后与模型组比较,柴胡生血方组、小柴胡合当归补血汤组EPO含量显著增高(P0.05)；(2)灌药1周后与模型组比较,小柴胡汤组SCF含量显著增高(P0.05)。其余各组间比较,差异无显著性(P0.05)。灌药2周后各用药组与模型组比较,差异无统计学意义(P0.05)。(3)灌药1周、2周后,与模型组比较,各灌药组IL-3含量均高于模型组,但差异无显著性意义(P0.05)。(4)灌药1周后与模型组比较,各用药组TGF-β1含量均降低,小柴胡合当归补血汤组、小柴胡汤TGF-β1含量显著降低(P0.05),而在灌药2周后与模型组比较,各灌药组TGF-β1含量均降低,但只有柴胡生血方组TGF-β1显著降低(P0.05)。(5)灌药1周后与模型组比较,小柴胡汤组TNF-α含量显著降低(P0.05)。灌药2周后,与模型组比较,当归补血汤组、小柴胡汤组、小柴胡合当归补血汤组、柴胡生血方组TNF-α含量显著降低(P0.05)。4.脑、心、肝组织病理统计结果：造模后,脑、心、肝组织病理指标中,只有肝脏组织病理指标统计有显著性差异(P0.01),灌药1周、2周后,与模型组比较,当归补血汤组、小柴胡汤组、柴胡生血方组、小柴胡合当归补血汤组病变程度及病变积分显著降低(P0.05)。结论：1.肝生血是肝应春生之气,有促生、催生及生发作用,能助他脏以生血。肝可以通过应生发之机,调节各脏腑生血功能,从而协助各脏腑完成血的生成。2.柴胡生血方对小鼠体重改善作用明显；在生血效应中,小柴胡汤表现为早期的促生血作用,小柴胡合当归补血汤生血效应持续,柴胡生血方在灌药第二周后生血效应明显且对促WBC成生效果强于其余用药组,当归补血汤生血效果不及小柴胡合当归补血汤与柴胡生血方。3.本造模方法得到的复合病因贫血小鼠模型对脑、心组织的病理影响不大,在组织缺血时,肝脏比心、脑组织敏感。在灌药后,几个补血药均对贫血小鼠肝脏病理缺血状态有所改善。4.小柴胡汤早期促生血作用机制可能与小柴胡汤率先促进干细胞分化早期细胞因子SCF表达、显著降低TNF-α表达有关；小柴胡汤、柴胡生血汤、小柴胡合当归补血汤可显著促进EPO、抑制TGF-β1、TNF-α表达。5.根据“肝生血气”理论确立的“调肝生血法”,在治疗血虚疾病方面具有有效性和可行性,为临床治疗贫血病症提供了新的思路,也为《内经》“肝生血气”理论提供了新的实验研究依据。
[Abstract]:Objective: the application of tiaogan hemogenesis method, observation of Minor Bupleurum Decoction and Danggui Buxue Decoction, Xiaocaihu, Danggui Buxue Decoction of Bupleurum Shengxue Recipe on blood loss and starvation effect model prepared by composite etiology of anemia mice, to explore the "Yellow Emperor", "Gan Sheng blood" theory, provide new ideas and methods for the clinical treatment of anemia, "Gan Sheng blood" provides an experimental basis theory. Methods: 1. of the "Chinese medical classics" in 1000 kinds of Chinese ancient medical books about "liver blood" theory analysis and analysis, summed up the "liver blood" theory. The selection of 2. SPF grade male and female feeding mice, the mice were randomly divided into blank group, model group, Xiaochaihu Decoction group, Danggui Buxue Decoction group, Xiaocaihu Danggui Buxue Decoction group and Chaihu Shengxue recipe group, compound model preparation causes anemia mice model, 14 day period. At the beginning of the fifteenth day, the treatment group began to fill the stomach with traditional Chinese medicine decoction, 7 days as the 1 course of treatment, with a total of 2 courses. To observe the change of blood deficiency mice body weight and body; after 2 weeks, 1 weeks, respectively, each group were randomly selected and 4 mice of each method by eyeball blood blood from peripheral blood, blood routine test, Elisa method was used to detect the serum cytokines SCF, EPO, IL-3, TGF-, beta 1 TNF-: the death from animal alpha in mouse liver, heart and brain to make pathological tissue sections to observe the pathological changes of brain, heart and liver tissue ischemia, and score statistics. Results: 1. weight change: before and after modeling, there are significant differences in body weight change of mice (P0.01); after 1 weeks, compared to the blank group and the model group, the perfusion group and model group, there was no significant difference in weight change (P0.05); after second weeks, compared with the control group compared with model group, weight less than the blank group, there was significant difference (P0.05), compared with the model group, Danggui Buxue Decoction group, Xiaocaihu Decoction group, Danggui Buxue Decoction group mice blood Bupleurum weight gain, with significant difference (P0.05). Compared with other groups, blank group, model group, Danggui Buxue Decoction group, Xiaochaihu Decoction group, Xiaocaihu Danggui Buxue Decoction group, 5 groups of mice weight were less than the weight of Bupleurum Shengxue prescription group, and the difference was significant (P0.05). 2. blood test results: compared with model group, each group of peripheral blood were improved, after 1 weeks, Xiaocaihu Danggui Buxue Decoction group, Xiaochaihu Decoction group, RBC mice blood HGB content was significantly higher than that in the model group (P0.05), after 2 weeks, Xiaocaihu together when return RBC Buxue Decoction group was significantly higher than that of model group (P0.05), Chaihu Shengxue prescription group HGB, WBC were significantly higher than those in the model group (P0.05). The detection results of cytokines in the serum of 3. indicators: (1) after 1 weeks compared with model group, Xiaochaihu Decoction group, Xiaocaihu Decoction group, Danggui Buxue Decoction group of Bupleurum blood concentration of EPO was significantly increased (P0.05); after 2 weeks compared with the model group, blood group, bupleurum Xiaochaihutang Danggui Buxue decoction group EPO were significantly increased (P0.05); (2) after 1 weeks compared with model group, Xiaochaihu Decoction group were significantly increased in SCF (P0.05). There was no significant difference between the other groups (P0.05). After 2 weeks of drug administration, there was no significant difference between the two groups (P0.05). (3) after 1 weeks and 2 weeks, the content of IL-3 in each group was higher than that in the model group, but there was no significant difference between the two groups (P0.05). (4) after 1 weeks compared with the model group, the treatment group TGF- beta 1 were lower, Xiaocaihu Danggui Buxue Decoction and Xiaochaihu Decoction group TGF- beta 1 was significantly lower (P0.05), and in 2 weeks after compared with the model group, the perfusion group TGF- beta 1 content decreased, but only the bupleurum Shengxue prescription group TGF- beta 1 was significantly decreased (P0.05). (5) compared with the model group for 1 weeks, the content of TNF- alpha in the small Bupleurum Decoction group decreased significantly (P0.05). After 2 weeks, compared with model group, Danggui Buxue Decoction group, Xiaochaihu Decoction group, Xiaocaihu Danggui Buxue Decoction group, bupleurum Shengxue prescription group TNF- alpha was significantly lower (P0.05). 4. brain, heart and liver tissue pathological results: model, brain, heart and liver tissue pathological indicators, only have statistically significant differences in liver tissue pathological index (P0.01), 1 weeks and 2 weeks, compared with model group, Danggui Buxue Decoction group, Xiaochaihu Decoction group, Chai Husheng blood Xiaocaihu Decoction group, Danggui Buxue decoction group and lesion severity score decreased significantly (P0.05). Conclusion: 1. liver blood liver should be Chunsheng gas, growth, birth and hair, can help him with dirty blood. The liver can regulate the function of blood producing in various viscera, thus assisting the viscera to complete the formation of blood. 2. Bupleurum Shengxue Recipe on mice body weight improved; in blood effect, Koshiba Koyu is promoting blood early role of Xiaocaihu Danggui Buxue Decoction for nourishing blood effect, bupleurum Shengxue prescription second weeks later in the blood effect on promoting and WBC effects in the treatment group the effect of Danggui Buxue Decoction, blood less than Xiaocaihu Danggui Buxue Decoction and Radix Shengxue recipe. 3. the model of the compound etiological anemia in mice has little effect on the pathology of brain and heart tissue, and the liver is sensitive to the heart and brain tissue during tissue ischemia. After the injection, several blood supplement drugs have improved the pathological ischemic state of the liver in the anemia mice. 4. Xiaochaihutang may Hematosis early promoting mechanism and Koshiba Koyu first to promote stem cell differentiation and early cytokine SCF expression significantly decreased expression of TNF- alpha expression; Xiaochaihu Decoction, Radix Bupleuri, Xiaocaihu Shengxue Decoction and Danggui Buxue Decoction can significantly promote EPO, inhibition of TGF- beta 1 and alpha TNF-. 5. according to the "Gan Sheng blood" established in the theory of "liver regulating method" is feasible and effective in the treatment of blood deficiency disease, provide new ideas for clinical treatment of anemia disease, but also for the "canon" and "Gan Sheng blood" theory provides a new experimental research
【学位授予单位】：成都中医药大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R259

【相似文献】