Ethanol extract of Nepeta angustifoia C. Y. Wu ameliorates hyperuricemia in fructose-induced mice.

As a popular medicinal plant traditionally used in Tibet of China, Nepeta angustifolia C. Y. Wu is mainly administered to treat apoplexia, cerebral haemorrhage, fainting and epilepsy and other symptoms, while its effect on hyperuricemia is still unclear. In the present study, we evaluated the improvement of the 70% ethanol extract of Nepeta angustifolia C. Y. Wu in fructose-induced hyperuricemic mice. The results revealed that Nepeta angustifolia C. Y. Wu significantly decreased blood glucose and blood lipid levels, as well as lowering the urinary levels of uric acid, creatinine and urea nitrogen. Meanwhile, it effectively restored the serum levels of uric acid, creatinine and urea nitrogen and inhibited serum and hepatic XOD activities and renal oxidative stress, while suppressing the secretions of TNF-α, IL-1ß and IL-6 in kidney. Nepeta angustifolia C. Y. Wu also attenuated the infiltration of inflammatory cells and reduced the production and accumulation of glycogen and collagen, while restoring the dysregulated protein expressions of renal URAT1, GLUT9, OAT1 and OAT3. In summary, our results support the idea that Nepeta angustifolia C. Y. Wu is a promising agent for treating hyperuricemia.

Malus toringoides (Rehd.) Hughes improves glucose and lipid metabolism and liver injury in high fructose-induced mice.

Malus toringoides (Rehd.) Hughes, as a traditional medicinal and edible plant used in Tibet, China, is used to treat hypertension, hyperlipidemia, and liver diseases. In recent decades, excessive fructose intake with diet has greatly increased the occurrence of a series of metabolic diseases including obesity, insulin resistance, hypertension, and hyperlipidemia. The present study was designed to investigate the effects of an ethanol extract of M. toringoides (EMT) on glucose and lipid metabolism and liver injury in high fructose-induced mice. The C57BL/6J male mice were orally administrated with 30% fructose solution for 8 weeks, and EMT was given orally for another 8 weeks. The level of liver lipids related parameters, hepatic oxidative stress, and inflammatory mediators was detected by the kits. The improving effects of EMT on liver injury and lipid accumulation of mice were observed by hematoxylin and eosin staining and Oil Red O staining. In vitro, the hypolipidemic effect of EMT on palmitic acid-induced HepG2 cells was detected by the kits and Oil Red O staining. Our results showed that EMT has the hypolipidemic effect in vivo and in vitro, and can improve liver injury caused by fructose intake though ameliorating oxidative stress and inflammatory responses. Thus, we suggested that EMT may be a candidate therapeutic agent to improve a series of metabolic diseases including obesity, insulin resistance, and hyperlipidemia. PRACTICAL APPLICATIONS: Our study was aimed to find a novel candidate drug for liver diseases using natural products. We assessed the protective effects of Malus toringoides (Rehd.) Hughes in the pathogenesis of glucose and lipid metabolism. In vivo, the plant significantly improved the disorder of blood lipid and blood glucose, and liver injury in mice induced by fructose, and in vitro, this plant significantly improved the lipid accumulation of HepG2 cells induced by palmitic acid. To sum up, our studies suggested that the plant may be beneficial in the prevention and management of diet-induced abnormal glucose and lipid metabolism and liver diseases. Therefore, it will be a candidate therapeutic agent to improve liver diseases.

Nepeta angustifolia attenuates responses to vascular inflammation in high glucose-induced human umbilical vein endothelial cells through heme oxygenase-1 induction.

ETHNOPHARMACOLOGICAL RELEVANCE: The traditional folk medicine Nepeta angustifolia C. Y. Wu (NA) reportedly possesses various biological activities, such as anti-inflammatory, analgesic, antihypoxia, and antifatigue effects. In this study, we evaluated the anti-vascular inflammation effects of N. angustifolia extract in human umbilical vein endothelial cells (HUVECs) induced by high glucose (HG) as well as the underlying mechanisms and verified its activity in diabetic rats. MATERIALS AND METHODS: HUVECs were exposed to 25 mM glucose to induce endothelial dysfunction. Adhesion molecule expression and reactive oxygen species (ROS) were assayed. IκB and IκB phosphorylation, nuclear factor-κB (NF-κB), HO-1 and nuclear factor erythroid 2-related factor 2 (Nrf2) were examined by Western blot. Nuclear localisation of Nrf2 was also examined using immunofluorescence. The in vivo study of NA was tested in diabetic rats in which the thoracic aorta and serum were collected to observe aorta histological change, and evaluate endothelial function and vascular inflammation. RESULTS: The results revealed that HG can significantly promote the generation of ROS, the expression of cell adhesion molecules (CAMs), and the phosphorylation and degradation of IκB and NF-κB activation in HUVECs. These HG-induced phenomena were suppressed by NA-induced heme oxygenase (HO)- 1 expression in a dose- and time-dependent manner by activating Nrf2. The HO-1 inhibitor tin protoporphyrin also dramatically reversed the NA-induced inhibition of CAM expression and the reduction in ROS production. Furthermore, NA also elicited anti-vascular dysfunction effects in diabetic rats, where endothelial function was improved and vascular inflammation was alleviated. CONCLUSION: All these findings indicated that NA attenuated high glucose-induced vascular dysfunction in vitro and in vivo.

Novel dual-function near-infrared II fluorescence and PET probe for tumor delineation and image-guided surgery.

Accurate tumor identification is essential in cancer management. Incomplete excision of tumor tissue, however, negatively affects the prognosis of the patient. To accomplish radical excision of tumor tissue, radiotracers can be used that target tumor tissue and can be detected using a gamma probe during surgery. Intraoperative fluorescence imaging could allow accurate real-time tumor delineation. Herein, a novel dual-modal imaging platform using base-catalyzed double addition of thiols into a propiolamide scaffold has been developed, allowing for the highly efficient and selective assembly of various thiol units in a protecting-group-free manner. The first small-molecule based αvß3-targeted NIR-II/PET probe 68Ga-SCH2 was concisely generated via this strategy and subsequently evaluated in mice bearing the U87MG xenograft. Excellent imaging properties such as good tumor uptake, high tumor contrast and specificity, tumor delineation and image-guided surgery were achieved in the small animal models. These attractive results of 68Ga-SCH2 allow it to be a promising αvß3-targeted NIR-II/PET probe for clinical translation.

Pyrazolopyrimidines as Potent Stimulators for Transient Receptor Potential Canonical 3/6/7 Channels.

Transient receptor potential canonical 3/6/7 (TRPC3/6/7) are highly homologous receptor-operated nonselective cation channels. Despite their physiological significance, very few selective and potent agonists are available for functional examination of these channels. Using a cell-based high throughput screening approach, a lead compound with the pyrazolopyrimidine skeleton was identified as a TRPC6 agonist. Synthetic schemes for the lead and its analogues were established, and structural-activity relationship studies were carried out. A series of potent and direct agonists of TRPC3/6/7 channels were identified, and among them, 4m-4p have a potency order of TRPC3 > C7 > C6, with 4n being the most potent with an EC50 of <20 nM on TRPC3. Importantly, these compounds exhibited no stimulatory activity on related TRP channels. The potent and selective compounds described here should be suitable for evaluation of the roles of TRPC channels in the physiology and pathogenesis of diseases, including glomerulosclerosis and cancer.

Hypolipidemic and Antioxidant Effects of Malus toringoides (Rehd.) Hughes Leaves in High-Fat-Diet-Induced Hyperlipidemic Rats.

Malus toringoides (Rehd.) Hughes (MT) leaves are traditionally used as a medicine for treating or preventing cardiovascular disease in Tibet. In addition to the effect of this medicinal plant on thrombosis, we tested its effect on dyslipidemia in a hypolipidemic rat model. A total of 60 healthy Sprague-Dawley rats were randomly divided into six groups, as follows: normal control, model control, simvastatin groups, and MT low-, medium-, and high-dose groups. The normal controls were fed with a normal diet, whereas all other groups were fed with a high-fat diet. After 6 weeks, the high-fat diet had induced hyperlipidemia in the rats, which were then orally administered with different doses of MT leaf extract (50, 100, and 200 mg/kg) for an additional 6 weeks. Serum levels of total cholesterol (TC), triglycerides (TG), low- and high-density lipoprotein cholesterol (LDL-c and HDL-c, respectively), as well as the antioxidant capacity of glutathione peroxidase (GSHP-x), superoxide dismutase (SOD), and malondialdehyde (MDA) were measured at the end of the study. MT significantly reduced serum TC, TG, and LDL-c and increased the HDL-c content in MT-treated rats compared with the model group. These changes were dose dependent. MT treatment also significantly elevated the activity of SOD and GSHP-x, and decreased the serum levels of MDA compared with untreated hyperlipidemic rats, thereby increasing serum antioxidant capacity. In addition, MT reduced liver steatosis in hyperlipidemic rats. Overall, MT exerts considerable hypolipidemic and antioxidant properties.

[Multi-component quantitative analysis combined with chromatographic fingerprint for quality assessment of Onosma hookeri].

A method for simultaneous determination of the shikonin, acetyl shikonin and ß, ß'-dimethylpropene shikonin in Onosma hookeri and the chromatographic fingerprint was estabished by HPLC-DAD on an Agilent Zorbax SB-column with a gradient elution of acetonitrile and water at 0.8 mL x min(-1), 30 degrees C. The quality assessment was conducted by comparing the content difference of three naphthoquinone constituents, in combination with chromatographic fingerprint analysis and systems cluster analysis among 7 batches of radix O. hookeri. The content of the three naphthoquinone constituents showed wide variations in 7 bathces. The similarity value of the fingerprints of sample 5, 6 and 7 was above 0.99, sample 2 and 3 above 0.97, sample 3 and 4 above 0.90, and other samples larger than 0.8, which was in concert with the content of three naphthoquinone constituents. The 7 samples were roughly divided into 4 categories. The results above indicated that the using of this medicine is complex and rather spotty. The established HPLC fingerprints and the quantitative analysis method can be used efficiently for quality assessment of O. hookeri.

Anti-hepatitis B virus active constituents from Swertia chirayita.

Four new compounds swertiachiralatone A (1), swertiachoside A (2), swertiachirdiol A (3) and swertiachoside B (4), together with twenty-six known ones were isolated from the ethanol extract of Swertia chirayita. Their structures were elucidated by extensive spectroscopic analyses (1D- and 2D-NMR, HRESIMS, UV, IR and [α]D). All compounds were evaluated for anti-hepatitis B virus (anti-HBV) activities on HepG 2.2.15 cells line in vitro, of which compounds 14 and 19 showed inhibitory activity on hepatitis B surface antigen (HBsAg) secretion with IC50 values of 0.31 ± 0.045 and 1.49 ± 0.033 mM; compounds 14 and 28 exhibited activity against hepatitis B e antigen (HBeAg) secretion with IC50 values of 0.77 ± 0.076 and 5.92 ± 1.02 mM; and eight compounds (8,9,13,14,24-26,29) possessed activity against HBV DNA replication with IC50 values of 0.07-0.33 mM. In particular (+)-cycloolivil-4'-O-ß-d-glucopyranoside (14) exhibited inhibition not only on the secretions of HBsAg and HBeAg with IC50 values of 0.31 ± 0.045 mM (SI=4.29) and 0.77 ± 0.076 mM (SI=1.75), respectively, but also on HBV DNA replication with an IC50 value of 0.29 ± 0.034 mM (SI=4.66).

[Effects of traditional tibetan medicine, Fructus Lonicerae microphyllae on phagecytosis and cytokines production of murine macrophages].

OBJECTIVE: To explore the effects of traditional Tibetan medicine, Fructus Lonicerae microphyllae (FLM) on phagecytosis and cytokines production of murine macrophages. METHOD: The phagecytosis of murine macrophages was analyzed by neutral red phagecytosis assay. The activities of IL-1 and TNF-alpha were measured by biological methods. The mRNA of TNF-alpha and INF-gamma expressed by macrophages was detected by RT-PCR. RESULT: The phagecytosis of murine macrophages was significantly enhanced by FLM at a concentration from 1 microg x mL(-1) to 100 microg x mL(-1) and the secretions of IL-1, and TNF-alpha from macrophages were markedly induced by FLM. Meanwhile, FLM also increased the expression of TNF-alpha mRNA and INF-gamma mRHA from macrophages in vitro. CONCLUSION: FLM could promote phagecytosis and cytokines production of murine macrophages.

[Immunosuppressive effect of Tibetan medicine, Artemisia vestita Wall Extract].

OBJECTIVE: To assess the immunosuppressive effect of Artemisia vestita Wall Extract (AV-ext) on mice. METHODS: The proliferative reaction of lymphocyte and the mixed lymphocytes reaction were used to determine the effects of AV-ext on the proliferation of mouse splenocyte in vitro and in vivo; Proliferative reaction of mouse splenocyte was used for detecting the effects of AV-ext on the level of IL-2 secreted by mouse activated splenocyte in vitro. Gelatin enzymogram method and adherence analytical method were employed to disclose the effects of AV-ext on mouse activated T-lymphocytes mobility and adherence. RESULTS: 1-100 microg/mL AV-ext exerted no obvious toxicity to mouse splenocyte, but it had obvious inhibitory effect on proliferative reaction of mouse splenocyte and mixed lymphocytes reaction induced by ConA. It also had obvious inhibitory effect on the level of IL-2 secreted by mouse activated splenocyte, on the production of MMP-9 by mouse activated T-lymphocytes, and on adherence. 150 mg/kg and 300 mg/kg of AV-ext, given to mouse per os for 7 days, could inhibit the proliferation of splenocyte and the secretion of MMP-9 by activated splenocyte of mouse. CONCLUSION: AV-ext can inhibit the cellular immune reaction of mouse obviously.

Aqueous extract from aerial parts of Artemisia vestita, a traditional Tibetan medicine, reduces contact sensitivity in mice by down-regulating the activation, adhesion and metalloproteinase production of T lymphocytes.

In the present paper, the effect of the aqueous extract from aerial parts of Artemisia vestita (AV-ext), a traditional Tibetan medicine, on ear contact sensitivity was examined. AV-ext significantly reduced the ear swelling when administered during the induction phase of picryl-chloride (PCl)-induced ear contact sensitivity in mice. The extract also showed a dose-dependent inhibition on lymphocyte proliferation and IL-2 production in Con A-activated spleen cells. The proliferation inhibition was confirmed in the mixed lymphocytes reaction. Furthermore, the adhesion of the isolated spleen cells from PCl-sensitized mice to type IV collagen was significantly decreased in a dose-dependent manner by AV-ext. Such decrease was also seen in AV-ext-treated Jurkat T cells and the T cells purified from above spleen cells. The purified spleen T cells from PCl-sensitized mice produced more matrix metalloproteinase-9 (MMP-9) than naive T cells, and AV-ext remarkably reduced MMP-9 production both in vivo and in vitro. These results suggest that AV-ext may alleviate contact sensitivity through blocking the activation of T lymphocytes and decreasing their localization to the inflammatory sites via down-regulating the potential of cell adhesion and metalloproteinase production.