强肌健力口服液对脾虚小鼠DNA合成的影响

Effect of qiangji jianli liquid on DNA synthesis in mice with spleen deficiency syndrome

目的:观察强肌健力口服液对实验性脾虚证小鼠DNA合成的影响,探讨该方防治脾胃虚损型重症肌无力的作用途径及中医脾肾相关的分子生物学基础。方法:实验于2005-06/07在广州中医药大学核医学实验室完成。选择健康雄性NIH小鼠69只,随机分为5组,即正常对照组(n=12)、脾虚模型组(n=15)、强肌健力口服液26g/kg组(n=14)、强肌健力口服液13g/kg组(n=15)及四君子汤组(n=13)。除正常对照组外,各组小鼠均采用利血平复制脾虚证动物模型,腹腔注射利血平0.2mg/(kg·d)。①正常对照组腹腔注射生理盐水0.1mL/(只·d)。②强肌健力口服液26g/kg,13g/kg组灌胃强肌健力口服液(由北芪、党参、升麻、白术、甘草等组成,每毫升含生药1.204g),给药剂量分别为26,13g/kg,1次/d。③四君子汤组灌服四君子汤(由党参、白术、茯苓、灸甘草组成)26g/kg,1次/d。各组均连续用药16d。给药结束后,测定各组小鼠脾、胸腺、肾、肝、小肠等组织DNA含量的变化;并测定各组小鼠造模前及用药结束后体质量和脾、肾、胸腺、肝脏质量/体质量比值的变化。结果:69只小鼠全部进入结果分析,无脱失。①用药16d后脾虚模型组脾脏、胸腺、小肠DNA含量均比正常对照组显著降低(q=3.45～4.96,P<0.05～0.01),而肾脏组织DNA含量则升高(q=3.81,P<0.05)。②用药16d后强肌健力口服液26,13g/kg组脾脏、胸腺DNA含量显著高于脾虚模型组[(20.73±4.44,12.30±4.61,7.37±4.43;12.76±2.12,11.58±2.46,8.41±3.86)DPM×104/g(q=3.22～12.43,P<0.05～0.01)];强肌健力口服液13g/kg组肾脏、肝脏DNA含量显著升高(q=3.50,3.76,P<0.05);而且体质量也显著高于脾虚模型组(q=9.98,9.94,P<0.01),脾脏、胸腺、肾脏、肝脏组织质量/体质量比值也显著高于脾虚模型组(q=3.53～7.41,P<0.05～0.01)。结论:脾虚证的发生与DNA合成减少有关,强肌健力口服液能促进DNA合成,从而发挥其健脾益气、强肌健力的作用,但发现强肌健力口服液26g/kg组能促进DNA合成,而13g/kg组则有抑制DNA合成的倾向,其调控途径有待于进一步研究。而脾肾相关理论有助于指导临床辨证施治。

AIM： To investigate effect of qiangji jianli liquid （QJJLL） on DNA synthesis in mouse models with spleen deficiency syndrome and explore the mechanism of treating myasthenia gravis in spleen and stomach deficiency symtrome and the possible relationship between spleen and kidney functions. METHODS： The experiment was completed at Laboratory of Nuclear Medicine of Guangzhou University of Traditional Chinese Medicine from June to July 2005. A total of 69 healthy male NIH mice were randomly divided into 5 groups： control group （n=12）, spleen deficiency model group （n=15）, 26 g/kg QJJLL group （n=14）, 13 g/kg QJJLL group （n=15） anti s ijunzi tang group （n=13）. Except mice in normal control group, model mice of spleen deficiency syndrome were established by injecting 0.2 mg/（kg .d） reserpine intravenously. ① Mice in normal control group were injected with 0,1 mL/（branch,day） saline intravenously. ② Mice in 26 g/kg QJJLL group and 13 g/kg QJJLL group were perfused with 26 g,/kg and 13 g/kg QJJLL respectively （consisted of beiqi, dangshen, shengma, baizhu, gancao, etc., 1.204 g raw mateirals） once a day, ③ Mice in sijunzi tang group were perfused with 26 g/kg sijunzi tang （consisted of dangshen, baizhu, fuling and jiugancao） once a day. The treatments in each group lasted for 16 days, After administration, changes of DNA content in spleen, thymus, kidney, liver and small intestine were measured, and alterations of body mass, ratios between body mass and weight of spleen, kidney, thymus and liver respectively were assayed before modeling and after administration. RESULTS： Totally 69 mice entered the final analysis without any loss, ① DNA contents in spleen, thymus and small intestine in spleen deficiency model group were lower than those in normal group 16 days after administration （q=3.45-4.96, P 〈 0.05-0,01）, but DNA content in kidney was increased （q=3.81, P 〈 0.05）. ② DNA contents in spleen and thymus in 26 g/kg QJJLL group and 13 g/kg QJJLL group were higher than those in spleen deficiency model group 16 days after administration [（20.73 ±4.44, 12.30±4.61, 7.37±4.43; 12.76±2.12, 11.58±2.46, 8.41±3.86） DPM ×10^4/g, q=3.22-12.43, P 〈 0.05-0.01]; DNA contents in kidney and liver in 13 g/kg QJJLL group were increased obviously （q =3.50, 3.76, P 〈 0.05）; body mass in 13 g/kg QJJLL group was also higher than that in spleen defieiency model group （q =9,98, 9.94, P 〈 0.01）; ratios between body mass and weight of spleen, thymus, kidney and liver respectively were increased as compared with those in spleen deficiency model group （q =3,53-7.41, P 〈 0,05-0.01）. CONCLUSION： The resuhs indicate that the development of spleen deficiency syndrome may be related to decrease of DNA synthesis. QJJLL can promote DNA synthesis so as to invigorate the spleen and replenish qi and strengthen muscle power. 26 g/kg QJJLL can promote DNA synthesis, but 13 g/kg QJJLL can inhibit DNA synthesis, and the regulation should be researched further. The theory of spleen-renal inter-relationship has its significance in directing determination of treatment based on differentiation of symptoms and signs.