针药结合对高尿酸血症肾损害大鼠尿酸及体内黄嘌呤氧化酶活性作用的研究

目的探索针刺结合中药复方矢志方对高尿酸血症肾损害大鼠尿酸及体内黄嘌呤氧化酶活性的作用。方法选用清洁级SD雄性大鼠50只,随机分为正常组、模型组、中药(矢志方)组、西药(别嘌醇)组、针药结合(针刺加矢志方)组,每组10只。予750μg·g-1·d-1氧嗪酸钾灌胃造模。造模后每日下午进行治疗干预,正常组及模型组灌胃2 ml蒸馏水,中药组予以矢志方灌胃,西药组予以别嘌醇灌胃,针药结合组予以矢志方灌胃加针刺治疗。针刺脾俞、肾俞、足三里、三阴交;隔日1次,每次留针30 min。连续4周后,检测各组大鼠血清尿酸、肌酐、尿素氮、血脂,24 h尿尿酸,24 h尿蛋白,血清及肝肾组织黄嘌呤氧化酶(XOD)活性。结果与正常组比较,模型组血尿酸、24 h尿尿酸水平、XOD活性明显升高,差异有统计学意义(P0.05),而血尿素氮、血肌酐差异无统计学意义(P0.05),说明该高尿酸血症模型成功建立。与模型组比较,中药组、西药组及针药结合组血尿酸显著降低,24 h尿尿酸总排泄量显著增加、血清及肝肾组织XOD活性显著降低,差异均有统计学意义(P0.05),说明3种治疗方法均有效;针药结合组与中药组组间比较,血尿酸、24 h尿尿酸、三酰甘油、胆固醇及血清、肝肾组织XOD差异均有统计学意义(P0.05),提示针药结合组相关疗效优于中药组。结论针药结合治疗可以有效降低高尿酸血症肾损害大鼠血尿酸的水平,并可促进尿尿酸的排泄,改善脂代谢水平,并可降低血清及肝肾组织XOD活性,可能为其降低血尿酸的机制之一。     

水蛭素对高尿酸血症大鼠尿酸盐转运体OAT1、URAT1、GLUT9表达的影响

目的 探究水蛭素对高尿酸血症大鼠的作用及机制。方法 雄性Wistar大鼠随机分为对照组、模型组、别嘌醇（30 mg/kg）组和水蛭素低、中、高剂量（0.2、0.4、0.8 g/kg）组。大鼠ig氧嗪酸钾（0.75 g/kg）,1次/d,连续5周,建立高尿酸血症模型;同时各给药组分别ig相应剂量的药物,1次/d,连续5周。采用生化法检测大鼠血清和尿液中的尿酸水平;免疫组化法测定肾组织中有机阴离子转运蛋白1（organic anion transporter 1,OAT1）水平;Western blotting法测定肾组织中葡萄糖转运体9（glucose transporter 9,GLUT9）、OAT1、尿酸盐转运体1（urate transporter 1,URAT1）蛋白表达;qRT-PCR检测肾组织中GLUT9、OAT1、URAT1 mRNA表达。结果 模型组大鼠血清和尿液中尿酸水平显著升高（P<0.01）,GLUT9、URAT1 mRNA和蛋白表达水平明显升高（P<0.01）,OAT1 mRNA和蛋白表达水平明显降低（P<0.01）;与模型组比较,水蛭素可显著降低大鼠血清和尿液中尿酸水平（P<0.01）,显著下调GLUT9、URAT1 mRNA和蛋白表达水平（P<0.01）,显著上调OAT1 mRNA和蛋白表达水平（P<0.01）。结论 水蛭素可通过调控高尿酸血症大鼠肾脏尿酸盐转运体OAT1、URAT1、GLUT9的表达,从而发挥降尿酸作用。     

Analgesic and anti-inflammatory activity of Lactuca sativa seed extract in rats

Lactuca sativa (Lettuce) is a member of Compositae family. In folk medicine of Iran, the seeds of this plant were used for relieving of inflammation and osteodynia. In this study, anti-nociceptive and anti-inflammatory activities of a crude methanol/petroleum ether (70/30, v/v) extract of the seeds have been evaluated. The extract exhibited a time- and dose-dependent analgesic effect in formalin test and also a dose-dependent anti-inflammatory activity in a carrageenan model of inflammation. The extract had no analgesic effect in tail-flick test up to the highest dose used (6 g/kg). No abnormal behavior and lethality was observed by the extract up to 6 g/kg. Preliminary phytochemical analysis showed the presence of triterpenoids, saponins and simple phenols in the extract.     

Juglone promotes antitumor activity against prostate cancer via suppressing glycolysis and oxidative phosphorylation

The treatments currently used for prostate cancer (PC) do not meet clinical needs, and thus, new therapies with greater effectiveness are urgently required. Metabolic reprogramming of tumor cells is emerging as an exciting field for cancer therapy. Although the Warburg effect is a common feature of glucose metabolism in many cancers, PC cells have a unique metabolic phenotype. Non-neoplastic prostate cells show reduced oxidative phosphorylation (OXPHOS) because large, accumulated zinc inhibits citrate oxidation. During transformation, there are low levels of zinc in PC cells, and the tricarboxylic acid (TCA) cycle is reactivated. However, metastatic PC exhibits the Warburg effect. Due to metabolic differences in prostate tissue, targeting metabolic alterations in PC cells is an attractive therapeutic strategy. In this study, we investigated the effect of juglone on energy metabolism in PC cells. We found that juglone inhibited cell proliferation and induced apoptosis. Mechanistically, we demonstrated that juglone suppressed OXPHOS and glycolysis due to its inhibition of hexokinase (HK), phosphofructokinase (PFK), and pyruvate kinase (PK) activity. Furthermore, downregulation of PFK and PK, but not HK contributed to the inhibition of these enzyme activities. The current study indicates that further development of juglone for PC treatment would be beneficial.     

Inhibitory effects of DA-9601 on ethanol-induced gastrohemorrhagic lesions and gastric xanthine oxidase activity in rats

The exposure of gastric mucosa to ethanol produces pathological changes such as inflammatory process, hemorrhagic erosions, even acute ulcers. The gastric mucosal lesions accompanied by a significant decrease of gastric blood flow and increase of reactive oxygen species (ROS) implicate a role of xanthine oxidase in ethanol-induced gastric hemorrhagic erosions. DA-9601, a novel antipeptic formulation of extracts of Artemisia asiatica Nakai, was studied for its inhibitory effect on gastric xanthine oxidase activity and type conversion of the enzyme that has a profound role in free radical generation. Intubation of absolute ethanol (4 g/kg) significantly induced gastrohemorrhagic lesions and lipid peroxidation in the rat stomach. Oral administration of DA-9601 at 40 mg/kg body weight significantly reduced ethanol-induced gastric mucosal hemorrhagic lesions and lipid peroxidation, which was proportional to the inhibitory effect of DA-9601 on alcohol-induced xanthine oxidase-type conversion and enzyme activity. The results suggest that alcohol-induced gastric mucosal damage may be, in part, due to the increased activity of xanthine oxidase and type conversion rate of the enzyme and that the preventive effect of DA-9601 on gastrohemorrhagic lesions would result from its inhibitory action against xanthine oxidase and oxidative stress in alcohol-treated rats.     

Effects of Biota orientalis extract and its flavonoid constituents, quercetin and rutin on serum uric acid levels in oxonate-induced mice and xanthine dehydrogenase and xanthine oxidase activities in mouse liver

The hypouricemic actions of Biota orientalis (BO) extract and its flavonoid constituents quercetin and rutin, were in vivo examined using oxonate-induced hyperuricemic mice. Quercetin and rutin, when administered three times orally to the oxonate-induced hyperuricemic mice, were able to elicit dose-dependent hypouricemic effects. The effects of quercetin and rutin were more potent than that of Biota orientalis extract at the same dose of 100 mg/kg. At doses of 50 mg/kg of quercetin or above, or at doses of 100 mg/kg of rutin or above, the serum urate levels of the oxonate-pretreated mice were not different from normal mice. In addition, Biota orientalis extract, quercetin and rutin, when tested in vivo on mouse liver homogenates, elicited significant inhibitory actions on the xanthine dehydrogenase/xanthine oxidase (XDH/XO) activities. The effects of quercetin and rutin resulted less potent than that of allopurinol. However, intraperitoneal administration at the same scheme did not produce any observable hypouricemic effect. These hypouricemic effects are partly due to the inhibition of XDH/XO activities in mouse liver. The pharmacological profile of the flavonoids is partly different from that of allopurinol. Such hypouricemic action and inhibition of the enzyme activity of quercetin and rutin may be responsible for a part of the beneficial effects of Biota orientalis extract on hyperuricemia and gout. The effects of quercetin and rutin on serum urate levels in hyperuricemic mice induced by oxonate and the inhibition of enzyme activities in mouse liver are discussed in relation to their absorption and metabolism, and their potential application to treat gout and hyperuricemia.     

Effects of acupuncture at Shu,Yuan, and Mu acupoints on blood serum uric acid and xanthine oxidase levels in a rat model of gout and hyperuricemia

OBJECTIVE: To investigate the effects of needling the Shu, Yuan, and Mu acupoints on serum uric acid(SUA), xanthine oxidase(XOD), and alkaline phosphatase(ALP) levels and the kidney index in a rat model of gout and hyperuricemia.METHODS: Fifty rats were randomly divided into five groups: blank, model, Shu-acupoint, Yuan-acupoint, and Mu-acupoint groups. A rat model of hyperuricemia was developed by intragastric administration of adenine and ethambutol. This experiment last for 90 d in total. Treatment groups underwent 3 courses of acupuncture. Each course involved a total of 10 interventions(one intervention every second day) with each intervention lasting15 min. There was a break for 10 d between courses. SUA and ALP were analyzed using an automatic biochemical analyzer and XOD was analyzed using immunofluorescence.RESULTS: Compared with the blank group, SUA and XOD levels in the model group were significantly higher and the renal index significantly improved. Compared with the model group, SUA and XOD levels in the three treatment groups decreased and the renal index significantly improved.When the three treatment groups were compared,the Mu-acupoint group showed the greatest decreases in SUA and XOD levels, followed by the Yuan-acupoint group. There was no significant difference in kidney index among the three treatment groups. There was no significant difference in ALP levels among the groups.CONCLUSION: The three treatments showed significantly reduced SUA and XOD levels compared with the control groups, possibly suggesting reduced renal damage.     

Plant cell culture extract of Cirsium eriophorum with skin pore refiner activity by modulating sebum production and inflammatory response

Facial pore enlargement is considered a significant esthetic and health concern in skincare cosmetics. The pores fulfill the critical function of keeping the skin surface hydrated and protected against microbial infections. The hyperseborrhea, the stress factors, and the hormonal triggers can cause pore size enlargement, causing higher susceptibility of the skin to microbe aggressions and inflammatory reactions. Thus, reducing excessive sebum production and keeping functional pores are two of the most requested activities in skincare cosmetics. A Cirsium eriophorum cell culture extract was investigated for its role in sebum regulation, stratum corneum desquamation, and anti‐inflammation. The extract was able to regulate essential markers associated with sebum secretion and pore enlargements, such as the enzyme 5α‐reductase, which plays a central role in sebum production, and the trypsin‐like serine protease Kallikrein 5, which promotes skin exfoliation and antimicrobial response. Moreover, the extract showed a sebum‐normalizing and pore refining activity in individuals having seborrheic or acne‐prone skins, suggesting a role of the C. eriophorum extract in rebalancing altered skin conditions responsible for pore enlargement.     

小远志中糖酯类成分及其体外抗黄嘌呤氧化酶活性研究

目的研究小远志Polygala sibirica var.megalopha中的糖酯类化学成分及其抗黄嘌呤氧化酶活性,并分析其构效关系。方法小远志75%乙醇提取物通过D101大孔树脂,依次用水和35%、65%、95%乙醇洗脱,采用尿酸生成法检测出65%乙醇洗脱部分抗黄嘌呤氧化酶活性最高(IC50=10.83μg/m L),对该部分采用多种柱色谱方法进行分离纯化,结合活性追踪,得到4个对黄嘌呤氧化酶具有高抑制活性的糖酯类化合物。结果 4个糖酯类化合物分别鉴定为远志寡糖酯A(1)、3,6′-二芥子酰基蔗糖(2)、3′-E-3,4,5-三甲氧基肉桂酰基-6-苯基蔗糖(3)、3′-E-3,4,5-三甲氧基肉桂酰基-4-苯基蔗糖(4),均为首次从该植物中分离得到。化合物1、3、4对黄嘌呤氧化酶的IC50分别为9.5、10.2、7.7μmol/L,比对照品别嘌呤醇(IC50=11.2μmol/L)抑制作用略强,而化合物2(IC50=1.3μmol/L)则显示出更强的抗黄嘌呤氧化酶活性,推测该类糖酯结构中苯乙烯基与糖基相连的结构片段是发挥药效的重要基团。结论糖酯类化合物作为新的黄嘌呤氧化酶高效抑制剂,具有开发成为抗痛风药的潜力。     

Effect of grape seed extract on lipid peroxidation, antioxidant activity and peripheral blood lymphocytes in rats exposed to x-radiation

The present studies were designed to evaluate supplemental grape seed extract (GSE) and vitamin E supplements on lipid peroxidation, on antioxidant systems and peripheral blood lymphocytes in rats exposed to x-rays. Three groups of rats were investigated: a control group (CG) received intraperitoneal (i.p.) physiological serum 1 mL/day (n=10), i.p.; a vitamin E group (VG) received 50 mg/kg/day (n=10); an i.p. grape seed extract group received 50 mg/kg/day (n=10). Four weeks later, a 6 Gy radiation dose was given to the rats. Blood samples were taken 24 h later after irradiation and lymphocyte, malondialdehyde (MDA), reduced glutathione (GSH), nitrate, nitrite, reduced ascorbic acid, retinol, beta-carotene and ceruloplasmin concentrations were analysed. The levels of GSH (p<0.05), retinol (p<0.001), beta-carotene (p<0.05) and ceruloplasmin concentration (p<0.001) in the GSE group were found to be higher than in the control group but the level of MDA (p<0.001) and nitrite concentration (p<0.05) in rats supplemented with GSE were found to be lower than in the control group. The results indicate that GSE enhanced the antioxidant status and decreased the incidence of free radical-induced lipid peroxidation in blood samples of rats exposed to x-radiation. The antioxidant effect of GSE given to animals was more effective than vitamin E administered before whole-body irradiation in rats.     

Inhibitory effect of hydro-methanolic extract of seed of Holarrhena antidysenterica on alpha-glucosidase activity and postprandial blood glucose level in normoglycemic rat

Ethnopharmacological relevance

The present experiment was conducted to search out the effect of hydro-methanolic extract of seed of Holarrhena antidysenterica on intestinal α-glucosidase activity in dose dependent manner and on the management of postprandial hyperglycemia in starch loaded rats.

Materials and methods

Activity of intestinal α-glucosidase was measured by in vitro method. Fasting blood glucose level was determined by single touch glucometer. Total phenol and flavonoids of seed extract of Holarrhena antidysenterica were estimated using gallic acid and quercetin standard curves, respectively.

Results

The degree of elevation in blood glucose level after starch administration was significantly (p < 0.05) less by the extract in respect to the control. The said extract also inhibited α-glucosidase activity having an IC₅₀ of 0.52 mg/ml. Phytochemical study revealed that the extract is rich in phenolic compounds (60.23 mg of gallic acid equivalent/g of extract) and flavonoids (360.23 mg of quercetin equivalent/g of the extract).

Conclusion

The extract exerts its antihyperglycemic effect by retarding the carbohydrate absorption from intestine through the inhibition in α-glucosidase activity and therefore resists postprandial hyperglycemia.     

Red ginseng extract inhibits the expression of MCP-1 and iNOS in Helicobacter pylori-infected gastric epithelial cells by suppressing the activation of NADPH oxidase and Jak2/Stat3

Ethnopharmcological relevance

Helicobacter pylori induced oxidative stress represents an important mechanism leading to expression of inflammatory mediators. Korean red ginseng is used in traditional medicine to inhibit inflammation. However, the anti-inflammatory mechanism of red ginseng is still under investigation. Thus, we investigated whether Korean red ginseng extract (RGE) inhibits NADPH oxidase, a source of reactive oxygen species (ROS), and the Jak2/Stat3 pathway, which mediates the expression of inflammatory mediators, in Helicobacter pylori-infected gastric epithelial cells.

Materials and methods

A standardized RGE was supplied by the Korea Ginseng Corporation. Human gastric epithelial cells (AGS) were treated with RGE and stimulated with Helicobacter pylori. NADPH oxidase activity, ROS levels, activation of Jak2/Stat3, and induction of MCP-1 and iNOS were determined.

Results

Helicobacter pylori infection resulted in an increase in ROS and activation of NADPH oxidase and Jak2/Stat3, which induced the expression of MCP-1 and iNOS in AGS cells. The induction of MCP-1 and iNOS was inhibited by both the Jak2/Stat3 inhibitor AG490 and RGE in Helicobacter pylori-infected cells. RGE suppressed NADPH oxidase activity by inhibiting translocation of cytosolic subunits p67phox and p47phox to the membrane and reduced ROS levels in Helicobacter pylori-infected cells.

Conclusion

RGE inhibits the expression of MCP-1 and iNOS by suppressing the activation of NADPH oxidase and Jak2/Stat3 in Helicobacter pylori-infected gastric epithelial cells.     

Danshen (Salvia miltiorrhiza) extract inhibits proliferation of breast cancer cells via modulation of Akt activity and p27 level

Danshen is widely used in traditional Chinese medicine, often in combination with other herbs. To check the effect of Danshen on the proliferation of breast cancer cells, Danshen extract was used to treat MCF‐7 and MCF‐7 HER2 cells, the latter of which overexpresses HER2. HER2 is a receptor tyrosine kinase, and is involved in signal transduction pathways leading to tumor cell proliferation. MTT and cell proliferation assays revealed that Danshen strongly inhibited the proliferation of both MCF‐7 vec cells and MCF‐7 HER2 cells. Flow cytometry analyses indicated that Danshen induced cell cycle delay in the G1 phase. HER2 expression was shown to confer resistance to Danshen‐induced inhibition of proliferation and cell cycle delay, suggesting that HER2 is responsible for the resistance to Danshen. Danshen treatment induced the down‐regulation of Akt phosphorylation and an increase in p27 in MCF‐7 vec and MCF‐7 HER2 cells. Nevertheless, MCF‐7 HER2 cells were more resistant to the Danshen‐induced inhibition of Akt phosphorylation and p27 up‐regulation. Copyright © 2009 John Wiley & Sons, Ltd.     

Evaluation of the anticonvulsant activity of the seed acetone extract of Ferula gummosa Boiss. against seizures induced by pentylenetetrazole and electroconvulsive shock in mice

Ferula gummosa Boiss. (Apiaceae) which has been used as an antiepileptic remedy in Iranian traditional medicine was evaluated for anticonvulsant activity against experimental seizures. The seed acetone extract of F. gummosa protected mice against tonic convulsions induced by maximal electroshock (the median effective dose [ED(50)]=198.3 mg/kg) and especially by pentylenetetrazole (ED(50)=55 mg/kg). Neurotoxicity (sedation and motor impairment) of the extract was assessed by the rotarod test and the median toxic dose (TD(50)) value of 375.8 mg/kg was obtained. Preliminary phytochemical analysis showed the presence of terpenoids and alkaloids in the extract. The acceptable acute toxicity of the extract recommends further studies to determine the mechanism(s) and compound(s) involved in the anticonvulsant activity.     

The hydroalcoholic extract of Cassia alata (Linn.) leaves and its major compound rhein exhibits antiallergic activity via mast cell stabilization and lipoxygenase inhibition

Ethnopharmacological relevance

Leaves of Cassia alata (family: Caesalpiniaceae) are ethnomedically claimed as anti-asthmatic. In the current study we aimed to investigate the anti-allergic activities of hydro-methanolic extract of Cassia alata (Linn.) and its constituents rhein and kaempferol on triple antigen/sheep serum-induced mast-cell degranulation in rats.

Materials and methods

Antiallergic activity of hydroalcoholic extract of Cassia alata along with its two components rhein and kaempferol was evaluated using in vivo mast cell stabilization assay. Inhibitory effect on lipoxygenase (LOX) enzyme was also evaluated in vitro. Further chemical standardization of Cassia alata extract was done using rhein and kaempferol by HPTLC-densitometric method.

Results

The hydroalcoholic extract of Cassia alata significantly inhibited mast cell degranulation at 200 mg/kg dose. Both chemical constituents rhein and kaempferol also showed potent (>76%) inhibition of mast-cell degranulation at 5 mg/kg. Extract and rhein inhibited LOX enzyme with IC₅₀ values of 90.2 and 3.9 μg/mL, respectively, whereas kaempferol was inactive.

Conclusion

Our results suggest that Cassia alata exhibit anti-allergic activity through mast cell stabilization and LOX inhibition. Thus, Cassia alata or its active constituents could be potential alternative treatment for allergic diseases.     

Huangkui capsule,an extract from Abelmoschus manihot (L.) medic,ameliorates adriamycin-induced renal inflammation and glomerular injury via inhibiting p38MAPK signaling pathway activity in rats

Ethnopharmacological relevance

Abelmoschus manihot (L.) medic (AM) is a natural medicinal plant used for the treatment of inflammatory diseases in China. Huangkui capsule (HKC), an extract from AM, has been proved clinically effective in improving renal inflammation and glomerular injury in chronic kidney disease (CKD). However, the dose-effects and the mechanisms involved in vivo are still unclear.

Aim of the study

This study was performed to examine the dose-effects of HKC on renal inflammation and glomerular lesion in adriamycin-induced nephropathy (ADRN), then to clarify the mechanisms in vivo of HKC by investigating its actions on modulating the activation of p38 mitogen-activated protein kinase (p38MAPK) signaling pathway.

Materials and methods

The rats with chronic ADRN, created by the unilateral nephrectomy and twice adriamycin injections (ADR, 4 mg/kg and 2 mg/kg) within 4 weeks, were divided into four groups, a Sham group, a Vehicle group, a high-dose HKC group, and a low-dose HKC group, and that, sacrificed at the end of the 4th week after the administration. The rat's general status, renal morphological appearance, proteinuria, blood biochemical parameters, glomerular morphological changes, podocyte shape, and macrophage (ED1⁺ and ED3⁺ cells) infiltration in glomeruli were examined, respectively. The protein expressions of inflammatory cytokines including tumor necrosis factor (TNF)-α and interleukin (IL)-2, as well as p38MAPK signaling molecules such as transforming growth factor (TGF)-β1, p38MAPK, and phosphorylated-p38MAPK (p-p38MAPK), were also evaluated individually.

Results

HKC at high dose of 2 g/kg/d not only significantly ameliorated the rat's general status, renal morphological appearance, proteinuria, albumin, and glomerulosclerosis, but also obviously reduced the infiltrated ED1⁺ and ED3⁺ macrophages in glomeruli and TNF-α protein expression in the kidney, in addition to these, evidently down-regulated TGF-β1 and p-p38MAPK protein expressions in ADRN rats, but had no influence on podocyte shape and renal function.

Conclusion

HKC could dose-dependently ameliorate renal inflammation and glomerular injury in ADRN rats, by way of reducing the infiltration and the activation of macrophages in glomeruli, and TNF-α protein expression in the kidney, as well as inhibiting p38MAPK signaling pathway activity via the down-regulation of p-p38MAPK and TGF-β1 protein expressions in vivo.