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。     

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。     

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.     

参附养心汤对老年阳虚水泛型慢性心力衰竭患者心功能及血清IL-6、TNF-α、hs-CRP水平的影响

目的 观察参附养心汤治疗老年阳虚水泛型慢性心力衰竭的临床效果。方法 将老年阳虚水泛型慢性心力衰竭患者92例随机分为观察组和对照组,每组46例。对照组采用西医常规治疗,观察组在对照组治疗方法基础上给予参附养心汤治疗。治疗4周后,对两组患者临床疗效、脑钠肽（BNP）、6 min步行距离、心功能指标[左心室舒张末期容积（LVEDV）、收缩末期容积（LVESV）、左心室射血分数（LVEF）]及细胞炎性因子[白细胞介素-6（IL-6）、肿瘤坏死因子（TNF-α）、超敏C-反应蛋白（hs-CRP）]进行比较。结果 治疗4周后,观察组总有效率为89.1%,显著高于对照组71.7%,差异具有统计学意义（P<0.05）;观察组的BNP水平显著低于对照组,而6 min步行距离显著高于对照组,差异具有统计学意义（P<0.05）;观察组的LVEDV、LVESV均显著低于对照组,而LVEF高于对照组,差异具有统计学意义（P<0.05）;观察组的血清hs-CRP、IL-6及TNF-α水平均显著低于对照组,差异具有统计学意义（P<0.05）。结论 参附养心汤能显著改善老年阳虚水泛型慢性心力衰竭患者的心功能指标,降低BNP水平,减轻炎症反应,提高运动耐量,疗效显著。     

附子-甘草对离体蛙心的影响及其指纹图谱研究

目的 研究附子-甘草合煎液和混合液对离体蛙心的影响及其物质基础。方法 通过离体蛙心实验观察附子-甘草合煎液和混合液对离体正常蛙心和心衰蛙心功能的影响,应用高效液相色谱法建立并测定附子-甘草煎液的HPLC指纹图谱。结果 附子-甘草合煎液与混合液的强心作用有差别;附子与甘草配伍后其指标成分的含量发生了明显变化,且合煎液与混合液的成分变化差别显著。结论 附子-甘草合煎液和混合液对离体蛙心的作用及其指纹图谱均不一致。     

Influence of Sini decoction on the proliferation and apoptosis of artery smooth muscle cells after balloon injury

AIM: To investigate the influence of Sini decoction (SND) on the proliferation and apoptosis of rabbit abdominal aorta smooth muscle cells after ballon injury and discuss the effect of vascular smooth muscle cell's (VSMCs) proliferation and apoptosis in post-percutaneous coronary intervention (PCI) restenosis (RS) and the feasibility of SND preventing post-PCI RS. METHODS: The animal model of rabbit abdominal aorta ballon injury was set up and the therapertic group was treated with SND. The shape of proliferative and apoptotic cell were investigated by electron microscope. Immunohistochemistry staining was performed using α-actin,PCNA and Cyclin E monoclonal antibodies. In situ Cell Death Detection Kit was used to identify apoptotic cells. Abdomial aorta angiography was operated in the 84th day subgroup and the stenosis degree was evalued by quantitative angiographic analysis. RESULTS: As compared with the control group, the therapeutic group displayed a lower proliferative percentage and a higher apoptosic percentage (P<0.05). Moreover, the apoptosic peek time was on the 14th day after operation,which was longer than the control group. CONCLUSION: SND effectly inhibited the proliferation of VSMCs and iuduced apoptosis in VSMCs.     

Effects of Sini decoction on the ultrastructure of small intestinal epithelial cells in rats with intestinal ischemia-reperfusion

AIM: To investigate the effect of Sini decoction (SND)on the ultrastructure of small intestinal epithelial cells in rats with intestinal ischemia-reperfusion. METHODS: Thirty-two Sprague-Dawley rats of both sexes were randomly divided into 4 groups: (1) Control group in which sham operation was performed; (2) Model group in which intestinal I/R was produced by clamping super mesenteric artery(SMA) for 1 hour and declamping SMA for 3 hours; (3) SND1 group in which SD (0.6 g/200 g rat) was given via stomach tube 3 d before intestinal I/R; (4) SND2 group in which SD (1.2 g/200 g rat)was given via stomach tube 3 d before intestinal I/R. A strip of small intestine was taken from distal end of ileum for electron microscopic examination. The two-dimensional structural parameters and three-dimensional structural parameters of mitochondria were calculated. RESULTS: (1)Morphological changes of small intestine: In control group, epithelial cells were orderly arranged, with normal mitochondria and intestinal villi. In model group, the gaps between epithelial cells widened. There were a lot of apoptotic cells. Microvilli were short and swelled. Mitochondria were swelled obviously with broken ridges. Endoplasmatic reticulum was severely dilated. In SND1 and SND2 groups, microvilli and epithelial cells were orderly arranged relatively, mitochondria was slightly swelled. (2) Structural parameters of mitochondria: In model group, there were the least mitochondria and the swelling of mitochondria was severe. In SND1 and SND2 groups, the mitochondria was more than that of model group and the swelling were slight. CONCLUSION: Sini decoction can protect small intestine from ischemia-reperfusion injury without dose-dependent effect.     

菊楂决明饮单煎和共煎对化学成分及抗氧化活性的影响

目的：考察菊楂决明饮中山楂、决明子（炒决明子）、菊花配伍单煎液与共煎液的化学成分及抗氧化活性差异。方法：采用紫外分光光度法测定总黄酮、总蒽醌的含量;采用酸碱滴定法测定总酸的含量;采用HPLC法测定山楂酸、大黄酚、橙黄决明素、绿原酸、木犀草苷、3,5-O-双咖啡酰基奎宁酸的含量。结果：山楂单煎液的总黄酮含量为 {1.63% }、总酸含量为5.44%、山楂酸含量为2.37%、对DPPH自由基的清除率为46.49%;决明子单煎液的总黄酮含量为0.54%、总蒽醌含量为0.27%、大黄酚含量为0.14%、橙黄决明素含量为 {0.52% }、对DPPH自由基的清除率为3.53%;炒决明子单煎液的总黄酮含量为0.52%、总蒽醌含量为0.29%、大黄酚含量为0.22%、橙黄决明素含量为0.56%、对DPPH自由基的清除率为25.07%;菊花单煎液的总黄酮含量为1.65%、绿原酸含量为 {0.99% }、木犀草苷含量为 {0.30% }、3,5-O-双咖啡酰基奎宁酸含量为2.65%、对DPPH自由基的清除率为45.89%;山楂、决明子、菊花共煎液总黄酮含量为2.62%、总酸含量为 {4.12% }、总蒽醌含量为0.25%、山楂酸含量为1.25%、大黄酚含量为0.16%、橙黄决明素含量为0.56%、绿原酸含量为 {3.01% }、木犀草苷含量为0.48%、3,5-O-双咖啡酰基奎宁酸含量为3.97%、对DPPH自由基的清除率为25.92%;山楂、炒决明子、菊花共煎液总黄酮含量为2.56%、总酸含量为 {3.94% }、总蒽醌含量为0.24%、山楂酸含量为0.43%、大黄酚含量为0.33%、橙黄决明素含量为1.00%、绿原酸含量为4.12%、木犀草苷含量为 {0.60% }、3,5-O-双咖啡酰基奎宁酸含量为4.74%、对DPPH自由基的清除率为44.51%。结论：山楂、决明子、菊花共煎后,与单煎液相比,总黄酮、大黄酚、橙黄决明素、绿原酸、木犀草苷、3,5-O-双咖啡酰基奎宁酸含量均升高;总酸、总蒽醌、山楂酸含量都有不同程度降低,对DPPH自由基清除率与单煎液相当。     

黄芪解毒汤对乳腺癌细胞Jag1基因和CyclinD1蛋白表达影响的研究

目的 探讨黄芪解毒汤抑制乳腺癌复发转移的机制是否与其干预Jag1基因和CyclinD1蛋白在人乳腺癌MCF-7细胞中的表达有关。方法 采用SPF级SD大鼠灌胃法制备黄芪解毒汤组、阿霉素组、参一胶囊组、生理盐水组的含药血清。运用细胞培养技术,MTT法检测发现最佳抑制浓度;以最佳抑菌浓度即20%浓度的含药血清干预人乳腺癌细胞,RT-PCR检测Jag1基因,Western-blot检测CyclinD1蛋白的表达。结果 各组与生理盐水血清组比较,Jag1基因和CyclinD1蛋白表达均明显降低（P<0.01）;与其他各组比较阿霉素血清组抑制作用最强（P<0.01）;黄芪解毒汤血清组与参一胶囊血清组比较差异无统计学意义（P>0.05）。结论 黄芪解毒汤可显著降低MCF-7细胞Jag1基因和CyclinD1蛋白表达,该结果可为黄芪解毒汤在乳腺癌术后治疗中的作用提供实验依据。     

瓜萎建白半夏汤对大鼠缺血再灌注 心肌细胞凋亡及Bcl-2 , Bax蛋白表达影响

目的研究瓜萎燕白半夏汤对缺血再灌注损伤大鼠心肌细胞凋亡及Bcl-2 , Bax蛋白表达影响〔方法通过 结扎大鼠冠状动脉左前降支造成心肌缺血再灌注损伤模型,各组动物至实验时限心肌缺血30 min、再灌注90 min后, 取出心脏、采用TUNEL检测心肌细胞凋亡,免疫组化方法检测心肌Bcl-2 , Bax蛋白表达、结果与假手术组对照,模型 组细胞凋亡率及Bax表达水平均明显升高、Bcl-2表达水平降低,组间比较差异有统计学意义(P<0.01);瓜萎燕白半夏 能有效降低Bax表达、升高Bcl-2表达水平,抑制细胞凋亡的发生,与模型组比较,差异有统计学意义(P<0.01)、结论瓜 萎燕白半夏汤可能通过上调Bcl-2 ,卜调Bax蛋白表达而有效抑制心肌缺血再灌注损伤大鼠心肌细胞凋亡的发生