Mechanism of Fuzilizhong decoction alleviating inflammatory damage of rats with non-alcoholic fatty liver disease

AIM To observe the effect of Fuzilizhong decoction on the inflammatory damage of non-alcoholic fatty liver disease （NAFLD） rats and to explore its mechanism. METHODS SPF male SD rats were randomly divided into 6 groups： control group, model group, high dose （20 mg·kg^-1·d^-1）, middle dose （10 mg·kg^-1·d^-1）, low dose （5 mg·kg^-1·d^-1） Fuzilizhong decoction group and Yishanfu （30 mg·kg^-1·d^-1）group, 8 rats in each group. A NAFLD rat modelwas established by intragastric administration of fat emulsion for 4 weeks. Then the drug was given for 4 weeks in each treatment group. HE staining was performed to observe the histopathological changes of the rat liver.The serum levels of interleukin-2（IL-2）, IL-6 and tumor necrosis factor-α（TNF-α） were measured by ELISA. The expression of toll like receptor 4（TLR4） and NF-κB p65 in liver tissues at mRNA and protein levels was determined by RT-qPCR and Western bolt,respectively. RESULTS Compared with control group, the inflammatory damage of liver tissue was more serious, the serum levels of IL-2, IL-6 and TNF-α, the mRNA expression TLR4 and NF-κB p65 in liver tissues were significantly increased in model group（P<0.05）. However, compared with model group, the liver pathological changes in each treatment group were significantly relieved, the serum levels of IL-2, IL-6 and TNF-α, the mRNA expression of TLR4 and NF-κB p65 in liver tissues were significantly reduced（P<0.05）.In addition, the changes of TLR4 and p-NF-κB p65 protein levels in liver tissue were consistent with the changes of TLR4 and NF-κB p65 mRNA. CONCLUSION Fuzilizhong decoction attenuates the inflammatory damages of NAFLD in rats by inhibiting TLR4/NF-κB p65 signaling pathway.     

超微粉碎对葛根芩连汤中葛根素、黄芩苷溶出的影响

将葛根芩连汤加工成细粉（40-65目）和超微粉（300目）,高效液相色谱法（HPLC）测定葛根芩连汤细粉和超微粉中主要有效成分葛根素、黄芩苷的含量,研究超微粉碎技术对葛根芩连汤主要有效成分葛根素、黄芩苷溶出的影响。结果葛根芩连汤细粉和超微粉中葛根素的含量分别为2.05%、3.20%;葛根芩连汤细粉和超微粉中黄芩苷含量分别为2.78%、3.94%。葛根芩连汤超微粉中的葛根素、黄芩苷含量比细粉分别提高56.10%、41.73%。表明超微粉碎技术能显著提高葛根芩连汤中主要有效成分葛根素和黄芩苷的溶出度。     

Effect of Jia-jian-yi-yin decoction on endothelial dysfunction in ovariectomized female rats

AIM: To investigate the effect of Jia-jian-yi-yin decoction on endothelium-dysfunction in ovariectomized rats. METHODS: The ovariectomized rats were treated with Jia-jian-yi-yin decoction or turbid liquid of estradiol valerate for 8 weeks. The vascular ring tension was measured. Scanning electron microscopy and Western blotting were applied to assess the function of endothelium-dependent dilation, aortic endothelial morphology and the expression of endothelial lipase. The pathologic sections were prepared to observe the effect of Jia-jian-yi-yin decoction on the uterus. RESULTS: In ovariectomized rats, the decrease in endothelium-dependent relaxation to acetylcholine (ACh) was reversed to normal level, the endothelial morphology returned to normal without lipid accumulation and the endothelial lipase expression was decreased by Jia-jian-yi-yin decoction. Furthermore, no obvious change of the wet weight of uterine between the ovariectomized rats with or without Jia-jian-yi-yin decoction treatment was observed. CONCLUSION: Jia-jian-yi-yin decoction may have protective effects on endothelium-dependent vasodilation and aortic endothelial morphology in estrogen-deficient animals.     

当归·黄芪及当归补血汤中元素形态分析

[目的]分析当归、黄芪及当归补血汤中的元素形态。[方法]采取中药常用煎煮方法对当归、黄芪及当归补血汤中Ca、Cu、Fe、Mg、Mn、Sr和Zn7种元素进行提取;用0.45μm滤膜分离煎液中的可溶态和不溶态;利用XAD-2大孔树脂将可溶态中元素分为有机态和无机态;采用ICP-AES法对各种形态中的7种元素进行测定。[结果]结果表明:当归、黄芪、当归补血汤药材及煎液中Ca、Fe、Mg、Zn的含量较高,7种金属元素的煎出率在8.77%~76.49%,煎液中各元素主要以可溶态形式存在,而在可溶态中又主要以无机态形式存在,当归、黄芪配伍共煎后对各元素形态变化并无规律性整体增加或减少。[结论]为进一步分析中药中金属元素的存在形态,深入探讨金属元素在中药药材、药效方面的作用机制提供了一定的理论基础。     

四君子汤对仔猪胸腺及脾脏生长激素受体mRNA表达的影响

选用90头新生"杜×长×大"三元杂交仔猪,随机分为3组,每组30头,分别为中药I组(中药添加剂量10 g/kg),中药Ⅱ组(中药添加剂量20 g/kg),对照组(基础日粮组),试验期为31 d.45日龄时每窝随机选取6头仔猪屠宰后,分别采集胸腺和脾脏.采用反转录聚合酶链式反应(RT-PCR)方法,定量分析仔猪胸腺及脾脏中生长激素受体(GHR)mRNA的相对丰度.结果显示,与对照组相比较,2种剂量中药组的仔猪胸腺及脾脏中生长激素受体mRNA的含量均不同程度地提高,中药Ⅱ组效果更为显著.     

绿茶抗大鼠肾间质纤维化作用的实验研究

采用"单肾切除加腺嘌呤灌胃法"建立大鼠慢性肾衰竭模型。治疗组予绿茶液灌胃治疗,对照组予尿毒清溶液灌胃治疗,连续治疗28天。取大鼠肾脏石蜡切片分别进行HE染色、PASM染色、Masson染色、Sirius Red染色、Mallory染色,光镜下观察大鼠肾组织5种染色的病理形态学改变,PASM染色、Masson染色、Sirius Red染色随机选择进行肾脏组织病理学图像分析。绿茶组在肾间质纤维化面积、肾小球系膜积分光密度值、肾间质中I、Ⅲ型胶原分布面积等方面的作用与模型组对比,P<0.05。说明绿茶能够减轻腺嘌呤对大鼠肾间质的病理损害,绿茶有抗肾间质纤维化的作用,并且能抑制I、Ⅲ型胶原纤维的增生,这可能是绿茶防治肾间质纤维化、延缓慢性肾衰竭发展进程的机制之一。     

实验性脾虚证大白鼠酶组织化学研究

将45只大白鼠随机分成3组，即实验组、治疗组及对照组，每组15只。以利血平致大白鼠脾虚证模型，分阶段捕杀动物，取肝脏、胃、十二指肠，用酶组织化学方法测定酸性磷酸酶(ACP)、碱性磷酸酶(ALP)、葡萄糖-6-磷酸酶(G-6-Pase)活性的变化。结果表明：实验组、治疗组、正常对照组各酶活性均发生了变化，在第7天、第14天时组内各酶存在着动态学上的变化。治疗组各酶活性接近正常对照组，说明四君子汤具明显的治疗作用。     

芪蓝抗毒饮治疗鸡传染性法氏囊病的剂量研究

比较了芪蓝抗毒饮三种剂量和中剂量三种给药次数对雏鸡实验性传染性法氏囊病的疗效。结果表明，中剂量和发病后该剂量每天给药1次、连续4天的疗效最好。     

Effects of Tian ma gou teng decoction on the serum free calcium concentration and the L-type calcium channels in the vascular smooth muscle cells in spontaneously hypertensive rats

AIM: The research was to investigate the effects of the Tian ma gou teng decoction on the electric physiology feature of L-type calcium channels in the vascular smooth muscle cells in spontaneously hypertensive rats (SHR), and to further explain the mechanism of the Tian ma gou teng decoction in the intervention of blood pressure.METHODS: 12-week-old SHRs were assigned randomly into five groups：group A (treated with Tian ma gou teng decoction), group B (treated with Tian ma gou teng decoction with subtraction concha haliotidis), group C (treated with nifedipine), group D (treated with concha haliotidis), group E (treated with normal saline as control), each group consisted of 9 rats. After treatments were conducted for 4 weeks, the free calcium concentration in serum was measured. The electric physiology feature of L-type calcium channels in the vascular smooth muscle cells was analyzed by patch clamp technique (PCT).RESULTS: No significant difference between group A and group C was observed in the serum free calcium concentration (P>0.05). There were significant differences among group B, group D and group E (P<0.05), compared to before treatment, the change in group E was the most obvious. A decrease in the L-type calcium channel current of vascular smooth muscle cells was observed in group A and group C. The function of group D was feeble, no decrease in the L-type calcium channels current of vascular smooth muscle cells was observed in group B and group E.CONCLUSION: Tian ma gou teng decoction can increase the serum free calcium concentration and block the L-type calcium channel current in vascular smooth muscle cells, indicating one of the mechanism of intervention of blood pressure.     

基于网络药理学分析白头翁汤治疗猪腹泻的作用机制

试验利用网络药理学研究方法尝试揭示白头翁汤治疗猪腹泻的活性成分、作用靶点及其基因功能、信号通路的机制。首先在中药系统药理学分析数据库中检索白头翁汤的所有化学成分、作用靶点,进而利用STRING、DAVID、NCBI数据库,Cytoscape软件构建化合物-靶点网络、蛋白质-蛋白质相互作用（PPI）网络、靶点-通路网络,研究白头翁汤治疗猪腹泻的作用机制。通过化合物的口服利用度和类药性筛选得出白头翁汤11个活性化合物,化合物-靶点网络结果显示,11个活性化合物含有63个相应靶点;白头翁汤作用于猪腹泻的PPI网络图包含45个靶点,主要靶点是雌激素受体1（estrogen receptor,ESR1）、CREB结合蛋白（CREB binding protein,CREBBP）、丝裂原活化蛋白激酶1（mitogen-activated protein kinase 1,MAPK1）、雄激素受体（androgen receptor,AR）等,与猪腹泻靶点基因直接相关的靶点是拓扑异构酶Ⅱβ（DNA topoisomerase Ⅱ,TOP2B）、ESR1、ESR2、糖皮质激素受体（glucocorticoid receptor,NR3C1）、AR和细胞核受体共激活剂2（nuclear receptor coactivator 2,NCOA2）;白头翁汤-猪腹泻PPI网络图关键靶点GO富集条目为5个,其中生物过程、分子功能、细胞组成相关的条目分别有3、1、1个;白头翁汤作用于猪腹泻PPI网络图KEGG信号通路有1条。白头翁汤可能主要通过金鱼草素、掌叶防己碱、8-异戊烯基二氢茆酚-7-葡糖苷、延胡索乙素、足叶草脂素、黄麻苷和8-羟基松脂醇等调控TOP2B、ESR1、ESR2、NR3C1、AR和NCOA2等靶点,基因功能富集于磷脂酶C激活G蛋白偶联受体信号通路、腺苷酸环化酶激活肾上腺素能受体信号通路、DNA模板转录、类固醇结合、细胞膜的组成部分,以及通过KEGG信号通路中神经活性的配体-受体相互作用信号通路来治疗猪腹泻。