目的:研究采用响应面法优化红景天抗氧化有效物质的提取工艺.方法:以乙醇浓度、提取时间、乙醇用量为主要影响因素,红景天苷提取率(％)、提取液体外DPPH清除率(％)和提取液体外总抗氧化能力为评价指标,采用3因素5水平响应面法对红景天抗氧化有效物质提取工艺进行优化.结果:Design-Expert软件优化得到的最佳提取工艺为,饮片加11倍体积46.60％乙醇溶液提取4.67 h,提取2次,合并提取液,浓缩,即得.该提取工艺下红景天苷提取率为1.12％,提取液DPPH清除率为89.13％,总抗氧化能力为1.39 mmol/L.结论:用响应面法优化得到的红景天抗氧化有效物质提取工艺简便易行,准确性和重现性好.

目的:观察希望液对衰老模型大鼠皮肤脂褐素沉积的影响及延缓皮肤衰老的药效.方法:采用吸入臭氧法复制大鼠衰老模型,各给药组分别灌胃给予阳性对照药(维生素E,13.5 mg·kg-1)及低、中、高剂量(225,450,900 mg·kg-1)希望液,模型组和正常对照组给予等量蒸馏水.检测大鼠血清总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-PX)、丙二醛(MDA)含量;HE染色显示皮肤病变,形态计量表皮角质层体积比;三氯化铁-铁氰化钾染色显示皮肤脂褐素,形态计量表皮脂褐素沉积体积比.结果:与正常对照组比较,模型组大鼠血清T-SOD、CAT、GSH-Px水平及皮肤表皮角质层体积比显著降低(P<0.05),MDA水平和脂褐素沉积显著升高(P<0.05或P<0.01).与模型组比较,希望液能升高大鼠血清T-SOD、CAT、GSH-Px水平及皮肤表皮角质层体积比,降低MDA水平和脂褐素沉积;其中希望液各剂量组的T-SOD、MDA水平及脂褐素沉积、中、高剂量组的CAT水平、高剂量组的GSH-Px水平及皮肤表皮角质层体积比与模型组比较差异有统计学意义(P<0.05或P<0.01).结论:希望液通过抑制氧化应激损伤,减少表皮脂褐素沉积,延缓皮肤衰老.

中医养生学是中华民族优秀传统文化的重要组成部分,历史悠久、源远流长.“养生”一词最早见于《庄子内篇·养生主》,是指通过精神调养、饮食调养、运动调养、起居作息调养等各种方法来保养身体的一种综合性强身益寿活动.

Objective:To develop a HPLC method for the determination of the concentration of Pae in rat plasma,and to study the pharmacokinetics of Pae in different stages of hepatic precancerous lesion model.Methods:Rats were successful modeling after given diethylnitrosamine(DEN),given single Pae by oral,blood taken from orbital at different time points,determined concentration of Pae by UPLC,the pharmacokinetic parameters were calculated by WinNonlin software;the expression of Estrogen Sulfate Transferase (EST) protein in liver and kidney were detected by immunohistochemistry.Results:Low dose group of Pae:from d000-d028-d056-d112,AUC,Cmax increased in turn,CL,V,MRT decreased in turn;middle dose group of Pae:from d000-d028-d056-d112,AUC,Cmax increased in turn,V,MRT decreased in turn,from d000-d028-d056,CL decreased in turn,from d056-d112,CL increased;high dose group of Pae:from d000-d028-d05 6-d112,AUC,Cmax increased in turn,CL,V,MRT decreased in turn.EST increased in liver endothelial cells and renal epithelial cells,decreased in liver cells.Conclusion:A simple and specific HPLC method for the analysis of Pae was successfully developed and applied to a pharmacokinetic study in rat plasma.

目的 探究杞红液对臭氧吸入联合冰水浴诱导大鼠血管纤维化的影响.方法 运用臭氧复合冰水浴造模法,制作寒凝血瘀衰老大鼠模型,检测血清NOS、MPO活力,ET-1含量;运用Masson染色和免疫组化来观察杞红液对动脉血管纤维沉积的影响.结果 与对照组比较,模型组NOS活力降低(P＜0.05),MPO活力升高(P＜0.05),ET-1含量降低(P＜0.05),血管纤维沉积增加(P＜0.05),TGF-β1表达量升高(P＜0.05).与模型组比较,杞红液低、中、高剂量对上述指标、胶原纤维沉积、TGF-β1表达量有不同程度改善(P＜0.05).结论 杞红液可能是通过调节大鼠NOS和ET-1的表达含量,调节机体应激反应,改善血管内皮功能及改善血管纤维化来实现对血管的保护作用.

Objective To discuss the influence of Qihong Fang (Wolfberry and Hawthorn Decoction) on heart in aged rats with cold-congealing and blood-stasis pattern.Methods SD rats (n =48) were divided randomly into normal control group,model group,vitamin E group (13.5 mg/kg),low-dose Qihong Fang group (45 mg/kg),mid-dose Qihong Fang group (90 mg/kg) and high-dose Qihong Fang group (180 mg/kg).The aged rat model of cold-congealing and blood-stasis pattern was established through 28-d ozone leading to aging and 14-d ice water leading to cold-congealing and blood-stasis pattern in other 5 groups except of control group.On the 29th d,the blood samples were collected from abdominal aorta for isolating serum and plasma,and heart was separated too.The vitalities of serum total superoxide dismutase (T-SOD),Cu-Zn SOD,Mn-SOD,catalase (CAT),glutathione peroxidease (GSH-PX),content of malondialdehyde (MDA),and prothrombin time (PT),activated partial thromboplastin time (APTT),thrombin time (TT) and fibrinogen (FIB) were detected.The morphologic changes of heart cells were observed after HE staining,fiber deposition quantity was observed after Masson staining,expression of transforming growth factor β1 (TGF-β1) was detected by using immunohistochemistry technique for reviewing aging model of cold-congealing and blood-stasis pattern,and protective effect of Qihong Fang on heart was observed in rats.Results Compared with normal control group,the vitalities of T-SOD,Cu-Zn-SOD,Mn-SOD,CAT and GSH-PX decreased significantly (P ＜ 0.05),MDA content increased significantly (P＜ 0.05,PT and APTT decreased significantly (P ＜ 0.05),and FIB content increased significantly (P ＜0.05) in model group.After Masson staining,morphometry observation by light microscope showed deposition of heart collagen fibers increased significantly (P ＜ 0.05),and expression of TGF-β1 increased significantly (P ＜ 0.05).Compared with control group,the vitalities of T-SOD,Cu-Zn-SOD,CAT and GSH-PX increased significantly (P ＜ 0.05),MDA content decreased significantly (P ＜ 0.05),PT and ATPP increased significantly (P ＜ 0.05),FIB content decreased significantly (P ＜ 0.05),deposition of heart collagen fibers decreased significantly (P ＜0.05),and expression of TGF-β1 decreased significantly (P ＜ 0.05) in mid-dose and high-dose Qihong Fang groups.Conclusion Qihong Fang has a protective effect on heart in aged rats with cold-congealing and blood-stasis pattern.

Objective To investigate the cardioprotective effect of Xiwang Ⅱ in mice exposed to passive smoking and its mechanism of action.Methods A total of 72 male Kunming mice were randomly divided into control group, model group, positive drug group, and low-, middle-, and high-dose Xiwang Ⅱ groups.The mice in the control group were exposed to laboratory air, and those in the model group were exposed to cigarette smoke for 45 consecutive days (3 hours a day).The drugs were administered since day 16 of exposure and the mice were sacrificed on day 45 to collect heart tissue.The level of malondialdehyde (MDA) in the heart was measured, as well as the activities of cardiac total superoxide dismutase (T-SOD),copper-zinc superoxide dismutase (Cu-ZnSOD), and myeloperoxidase (MPO).The histomorphological features of the heart were observed and the collagen fiber area in the heart was calculated.Immunohistochemistry was used to measure the expression of transforming growth factor-β1 (TGF-β1) in the heart.Results Compared withthe control group, the model group had significant reductions in the activity of Cu-ZnSOD and T-SOD (P<0.05) and significant increases in MPO activity, MDA level, cardiac muscle fibers, and TGF-β1 expression (all P<0.05).Compared with the model group, the low-, middle-, and high-dose Xiwang Ⅱ groups had significant increases in the activities of Cu-ZnSOD and T-SOD (P<0.05) and significant reductions in MPO activity, MDA level, cardiac muscle fibers, and TGF-β1 expression (all P<0.05).Conclusion Xiwang Ⅱ exerts its cardioprotective effect on mice exposed to passive smoking by regulating oxidative stress response in the heart.

Objective:To establish an analytical method for determination of Pae in tissues of hepatic precancerous lesion model rats by UPLC,and study the tissue distribution of Pae in rats.Methods:The concentration of Pae in rats tissues were determined by UPLC.Results:In control group,kidney had the highest tissue concentration of Pae,followed by stomach,small intestine,brain,liver,spleen,heart,lung;in model group,liver had the highest tissue concentration of Pae,followed by kidney,spleen,small intestine,stomach,heart,brain,lung.And its AUCall in model group was larger than that in control group.Conclusion:Compared with the control group,the content of Pae in model group increased.

Preparations of Chaihu Guizhi Ganj iang Tang prepared with different extraction technology are given to model rats through intragastric administration,and primary data including seven indexes on hepatic function and lipid peroxidation are collected to investigate their protective effects on hepatic inj ury.All the experimental data are conducted through grey relational analysis to find out that in three different technologies, the preparation of chahu Guizhi Ganj iang Tang has the best therapeutic effect and the optimum administration time (about three months)for rats.

Objective To study the impact of xiwang Ⅱ on the liver of passive - smoking mice. Methods Seventy - two Kunming male mice were randomized into a control group,a model group,a procya-nidin group,the xiwang low - dose group,the xiwang middle - dose group and the xiwangⅡ high - dose group. In the control group,the mice were exposed to the natural air and the models were prepared in the rest groups. Medication was given since the 16th day after modeling in each group. The double distilled water was used for lavage every day in the control group and the model group,5. 49 ml/ kg. In the procyanidin group, procyanidin,16. 47 ml/ kg was used every day for lavage. In the low - dose,middle - dose and high - dose groups of xiwang Ⅱ,xiwang Ⅱwas used at 1. 83 ml/ kg,5. 49 ml/ kg and 16. 47 ml/ kg separately every day for lavage. The tissues were collected in 45 days. Serum was collected from abdominal aorta and liver was ob-tained. CAT activity,the levels of AST and ALT in serum,T - SOD and Cu/ Zn - SOD activities and MDA were detected. The liver histology was observed. The volume constituent ratio of hepatocytes and volume ratio of hepatic collagen fibers were calculated. Results Compared with the control group,the activities of Cu/ Zn- SOD,T - SOD and CAT were reduced(P < 0. 01),the levels of MDA,AST and ALT were increased(P <0. 01),the volume constituent ratio of hepatocytes and volume ratio of hepatic collagen fibers were increased (P < 0. 01)in the model group. As compared with the model group,the activities of Cu/ Zn - SOD,T - SOD and CAT were increased apparently(P < 0. 01),the levels of MDA,AST and ALT were reduced apparently (P < 0. 05,P < 0. 01),the volume constituent ratio of hepatocytes and volume ratio of hepatic collagen fibers were reduced(P < 0. 01)in the low - dose,middle - dose and high - dose groups of xiwang Ⅱ. Conclusion Xiwang Ⅱprotects the liver from damage in the passive - smoking mice probably through improving the level of antioxidant enzymes,regulating oxidative stress,maintaining anti - oxidative balance,alleviating lymphocyte infiltration,preventing hepatocyte edema and delaying liver fibrosis.

Objective To evaluate the protection effect of Hope I decoction on ozone inhalationinduced injury in rats. Methods 48 male SD rats were randomly divided into 6 groups,i. e.,control,model,VE( 0. 0135 g / kg),Hope I decoction at three dosages( 0. 045,0. 090,0. 180 g / kg respectively).Modeling rats were exposed to ozone in an exposure cabinet( 10 h / d × 45d),then some of them were intragastrically administrated with vitamin E or Hope I decoction. Then,the level of SOD,LPO,CAT and GSHPx in rat serum were detected and pathologic change of lung tissue were observed,Immunohistochemistry for surfactant protein A( SP-A) content were observed. Results The activity of SOD,CAT and GSH-Px in serum were increased and LPO level were reduced in all Hope I groups. Histopathological slides of these three groups also showed an attenuated extent of alveoli dilatation and collapse induced by ozone exposure,surfactant protein A of these three groups were significantly enhanced. Conclusion Hope I decoction hasgreat protection effect on ozone inhalation-induced injury in rats.

目的 采用星点设计-响应面法优化温胆汤提取工艺.方法 以乙醇浓度、提取时间、乙醇用量为主要影响因素,以总酸和橙皮苷含量为评价指标,采用3因素5水平星点设计-响应面法对温胆汤乙醇提取工艺进行优化;以提取时间和加水量为主要影响因素,总多糖为指标,采用2因素5水平对温胆汤水提取工艺进行优化.结果 最佳提取工艺为饮片先加入10.73倍量60％乙醇,提取2次,每次提取1.68 h,收集乙醇提取液;滤渣加入8.13倍量水,提取2次,每次提取1.34h;合并乙醇和水提取液,浓缩即得.结论 用星点设计-响应面法优化得到的温胆汤提取工艺简便易行,准确性和重现性好.

目的：研究丹皮酚（Pae）对二乙基亚硝胺（DEN ）诱发大鼠肝癌前病变的预防作用，并探讨其作用机制。方法：采用腹腔注射DEN 诱发大鼠肝癌前病变模型。雄性w istar大鼠按照体质量随机分成3组：对照组、模型组、丹皮酚组。造模给药16周后处死大鼠，观察各组大鼠一般状况，进行肝脏病理组织学检查，检测肝功能指标ALT、AST、GST、TBIL ，脂质过氧化指标LPO、SOD、INOS、GSH‐PX。结果：病理组织学显示，丹皮酚可以显著降低肝细胞异型增生灶和异型增生结节数目；降低血清中异常升高的ALT、AST、TBIL、GST、INOS、LPO水平，升高血清中降低的SOD、GSH‐PX水平。结论：丹皮酚对于大鼠肝癌前病变具有一定程度的预防保护作用，其保护机制可能与改善肝功能，提高抗氧化能力，抑制脂质过氧化反应有关。

Objective To study the impacts of compound yanshou decoction on the liver in the rats of senility and hyperlipidemia. Methods Sixty male SD rats were randomized into a control group(standard fodder + nature air),a model group(high - fat fodder + 4 h ozone contamination),a positive medicine group (fenofibrate,18. 0 ml/ kg),and the low - dose,middle - dose and high - dose groups of compound yanshou decoction(4. 5 ml/ kg,9. 0 ml/ kg,18. 0 ml/ kg). Medication started on the 35th day and in 70 days of contam-ination. Serum was collected from abdominal aortal blood. The levels of SOD,LPO,GSH - PX,TG,TC,LDL- C and HDL - C were detected. The damage degree of liver tissue was observed pathologically. The in situ content of oxidase was measured morphologically. Results In the model group,the activity of SOD and GSH- PX and the level of HDL - C were reduced(P ﹤ 0. 05 or P ﹤ 0. 01),and the levels of LPO,TG,TC and LDL were increased(P ﹤ 0. 05,P ﹤ 0. 01);the hepatocyte edema,fatty degeneration and collagenous fiber sedimentation happened and NOX2 in situ content was increased. In the low - dose,middle - dose and high -dose groups of compound yanshou decoction,the improvements were achieved in the above biochemical inde-xes,collagenous fiber sedimentation and NOX2 in situ content. Conclusion Compound yanshou decoction acts on the protection to the liver of rat with ozone - induced senility and hyperlipemia.

目的 采用多重指标综合优化复方柴胡颗粒的提取工艺,并制备其颗粒剂.方法 综合以柴胡皂苷a、柴胡皂苷d、芦丁、橙皮苷和浸膏量为检测指标,采用L9(34)正交设计实验,考察乙醇浓度、溶剂用量、提取时间以及提取次数对复方柴胡颗粒提取效果影响,优化提取工艺并进行验证,以得到的干膏为原料,单因素考察稀释剂糊精和矫味剂阿斯巴甜的用量,制备复方柴胡颗粒.结果 最佳提取工艺为10倍量80％乙醇每次提取2h,提取3次;干膏与糊精比例为1∶0.9,润湿剂为85％乙醇,阿斯巴甜用量为1.0％时得到复方柴胡颗粒较好.结论 多指标综合考察复方柴胡颗粒最佳提取工艺,验证实验表明该工艺可行、重复性良好,单因素考察复方柴胡颗粒制剂制备工艺,得到外观优良、口感较佳的颗粒剂.

Objective:To establish the quality control method for compound Jieyu granules. Methods:Liquorice was identified by TLC. saikosaponin a, Saikosaponin d and rutin were determined by HPLC. Results:The spots on TLC plates were clear without any in-terference. The linearity was achieved within the range of 0. 508-16. 200 μg (r=0. 999 8) for saikosaponin a, 0. 503-16. 100 μg(r=0.999 7) for saikosaponin d, and 0.130-4.250 μg(r =0.999 9) for rutin. The average recovery was 99.7%(RSD =2.03%), 99. 8%(RSD=1. 44%) and 102. 6%(RSD=1. 40%), respectively. Conclusion:The method is simple, reliable and accurate, and can be applied as the quality control method for compound Jieyu granules.

Objective:To investigate the protective effects of baicalin on liver in hepatic precancerous mice induced by diethylnitrosamine(DEN).Methods:Hepatic precancerous model was obtained by administration of DEN through intraperitoneal injection to male mice,and the pharmacologic effects of baicalin were assayed by histopathologic examination and levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),gultathione-S-transferases(GST) and alkaline phosphatase (ALP).Results:The liver functions were serious damaged by administration of DEN,which showed up as morphologic changes and abnormal structure in model mice,such as hver cirrhosis,hepatic fibrosis which divided the liver lobule,liver cell denaturation, hyperplasia,intranuclear inclusions and tendency to atypia,and heavy local infiltration by macrophages was observed,at the same time,enzymatic activities in serum were increased abnormally(P0.05).Situations of mice treated by baicalin were much better,and hepatic fibrosis was decreased,which was failed to divide the liver lobule,and scarce liver cell denaturation,atypical hyperplasia,intranuclear inclusions could be observed.Enzymatic activities in serum were decreased markedly compared with those in model mice(P0.05).Conclusion:Baicalin is partly effective to protect the liver functions in hepatic precancerous mice induced by DEN.

目的:研究二乙基亚硝胺诱发大鼠肝癌前病变的时效关系与量效关系.方法:Wistar♂大鼠100只,体质量均衡随机分5组；每84 h 1次分别腹腔注射二乙基亚硝胺0、25、50、75、100 mg/kg造模.每组体质量排序分10小组,年龄配对；造模时程最短0d,间隔14d延长,最长126 d,灌注固定肝脏.常规石蜡包埋,6 μm切片,HE染色,观察病变性质后显微照相；使用Image-Pro Plus软件,分别形态计量10、20、40倍物镜下汇管区(R1)、肝索(R2)、肝细胞核(R3)的体积构成比,观察每10个高倍镜视野下的核分裂像；计算各动物异型性指数[Ei=(1-R1)-1·(R3)·(R2)-1].Prism 4软件回归“二乙基亚硝胺导致异型性指数”的时效曲线及曲线下面积,进而导致曲线下面积的量效关系.结果:以异型性指数(肝细胞核/浆体积比校正值),回归0、25、50、75、100 mg/kg二乙基亚硝胺造模的半效时程(95％CI)分别为70 347(0-)、1 734(937-3 211)、1 536(948-2 490)、1 530(890-2 632)、1 183(955-1 466)h,曲线下面积分别为0.0064、0.0084、0.0123、0.0165、0.0167[异型性指数·log(h)].以曲线下面积,回归二乙基亚硝胺诱发肝细胞癌前病变的半效剂量为48.255mg/kg.50.000mg/kg造模各时程的异型性指数与核分裂像总数之间呈直线正相关(y=0.0023x-0.0056,r=0.9217,n=10,P＜0.01).结论:以异型性指数为指标,每84 h腹腔注射1次二乙基亚硝胺,诱发♂大鼠肝细胞癌前病变的最佳造模剂量为48.255 mg/kg(约50.000mg/kg),最佳造模累积连续时程为64 d.核分裂像计数验证了异型性指数的临床预警价值.