Natural Xanthine Oxidase Inhibitor 5-O-Caffeoylshikimic Acid Ameliorates Kidney Injury Caused by Hyperuricemia in Mice.

Xanthine oxidase (XOD) inhibition has long been considered an effective anti-hyperuricemia strategy. To identify effective natural XOD inhibitors with little side effects, we performed a XOD inhibitory assay-coupled isolation of compounds from Smilacis Glabrae Rhizoma (SGR), a traditional Chinese medicine frequently prescribed as anti-hyperuricemia agent for centuries. Through the in vitro XOD inhibitory assay, we obtained a novel XOD inhibitor, 5-O-caffeoylshikimic acid (#1, 5OCSA) with IC50 of 13.96 µM, as well as two known XOD inhibitors, quercetin (#3) and astilbin (#6). Meanwhile, we performed in silico molecular docking and found 5OCSA could interact with the active sites of XOD (PDB ID: 3NVY) with a binding energy of -8.6 kcal/mol, suggesting 5OCSA inhibits XOD by binding with its active site. To evaluate the in vivo effects on XOD, we generated a hyperuricemia mice model by intraperitoneal injection of potassium oxonate (300 mg/kg) and oral gavage of hypoxanthine (500 mg/kg) for 7 days. 5OCSA could inhibit both hepatic and serum XOD in vivo, together with an improvement of histological and multiple serological parameters in kidney injury and HUA. Collectively, our results suggested that 5OCSA may be developed into a safe and effective XOD inhibitor based on in vitro, in silico and in vivo evidence.

Artificially Cultivated Ophiocordyceps sinensis Alleviates Diabetic Nephropathy and Its Podocyte Injury via Inhibiting P2X7R Expression and NLRP3 Inflammasome Activation.

BACKGROUND/AIMS: It is known that chronic low-grade inflammation contributes to the initiation and development of both diabetes and diabetic nephropathy (DN), so we designed this study to investigate the role of P2X7R and NLRP3 inflammasome in DN pathogenesis and the antagonistic effects of artificially cultivated Ophiocordyceps sinensis (ACOS). METHODS: A rat model of DN caused by high-fat-diet feeding and low-dose streptozotocin injection and a mouse podocyte injury model induced by high-glucose (HG) stimulation were established, and the intervention effects of ACOS on them were observed. The biological parameters of serum and urine and the pathological manifestations of kidney tissue were examined. The expression of mRNA and protein of P2X7R and NLRP3 inflammasome (NLRP3, ASC, and caspase-1) and downstream effectors (IL-1ß and IL-18), as well as podocyte-associated molecules, was determined by real-time quantitative PCR and Western blot assay, respectively. RESULTS: The DN rats showed to have developed insulin resistance, elevated fasting blood glucose, increased urinary protein excretion, and serum creatinine level as well as corresponding glomerular pathological alterations including podocyte damages. ACOS significantly antagonized the above changes. The experiments in vivo and in vitro both displayed that the mRNA and protein expression of P2X7R, NLRP3, ASC, caspase1 (procaspase-1 mRNA in the gene level and active caspase-1 subunit P10 in the protein level), IL-1ß, and IL-18 was significantly upregulated and the mRNA and protein expression of podocyte-associated molecules was significantly changed (downregulation of nephrin, podocin, and WT-1 expression and upregulation of desmin expression) indicating podocyte injury in the kidney tissue of DN rats and in the HG-stressed mouse podocytes, respectively. ACOS also significantly antagonized all the above changes. CONCLUSION: Our research work suggests that P2X7R and NLRP3 inflammasome are involved in the pathogenesis of DN, and ACOS can effectively inhibit the high expression of P2X7R and the activation of NLRP3 inflammasome, which may contribute to the therapeutic effects of Ophiocordyceps sinensis.

Pharmacokinetics of the main compounds absorbed into blood after oral administration of Liu Wei Di Huang Wan, a typical combinatorial intervention of Chinese medical formula.

Liu Wei Di Huang Wan (LW) has been used as an active Chinese patent formula for "Five Late Syndrome" of Children for thousands of years. Due to the complexity in its chemical constituents, the pharmacokinetics of this formula have not been elucidated clearly, and the understanding of its pharmacological properties has been delayed. Previous studies have identified the constituents absorbed into blood after the oral administration of LW; moreover, 5-hydroxymethyl-2-furoic acid (HMFA), loganin and paeonol have been proved as surrogate markers. In this study, a rapid validated high-performance liquid chromatography method was developed for determining three marker compounds in plasma. The analysis was performed on a Waters Symmetry Shield™ RP(18) column with acetonitrile and 0.15% phosphoric acid as the mobile phase, which showed acceptable linearity, intra- and inter-day precision, and accuracy. By using the established method, the pharmacokinetic analysis of LW was carried out. The t (1/2)α and t (1/2)ß were 2.62/32.66, 0.46/4.71 and 1.30/23.51 h and the climax times and concentrations were 0.56/683.75, 0.70/2826.11 and 0.62 h/4030.48 ng ml(-1) for HMFA, loganin and paeonol, respectively. Especially, both the absorption and disposition of HMFA were swift (t (1/2) kα 0.1 h, t (1/2)α 2.62 h), but the elimination was quite slow (t (1/2)ß 32.66 h); this phenomenon reflected the synergetic effect of LW combinatorial intervention and the value of compatibility can be more clearly understood. The pharmacokinetic characters of HMFA, loganin and paeonol not only elucidated the steady and long-lasting pharmacological properties, but they also revealed the practical value of the compatibility of Chinese medical formula.

Protective effects of sweroside on human MG-63 cells and rat osteoblasts.

Herbal Fructus Corni is a folk medicine with a long history of safe use for treating osteoporosis in postmenopausal women or elderly men in Asia. Sweroside is a bioactive herbal ingredient isolated from Fructus Corni, which has been widely used for the treatment of osteoporosis in traditional Chinese medicine (TCM). Unfortunately, the working mechanisms of this compound are difficult to determine and thus remain unclear. The aim of the study was performed to determine the potential molecular mechanism of the anti-osteoporotic effect of sweroside on the human MG-63 cells and rat osteoblasts. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test was used to observe the effect of sweroside on cell proliferation. The activity of alkaline phosphatase (ALP) and the amount of osteocalcin were also assayed the cell differentiation. Sweroside significantly increased the proliferation of human MG-63 cells and rat osteoblasts (P<0.01). It increased the activity of ALP, and osteocalcin was also elevated in response to sweroside (P<0.05). Of note, flowcytometer assay showed that sweroside can attenuate and inhibit apoptosis. Sweroside has a direct osteogenic effect on the proliferation and differentiation of cultured human MG-63 cells and rat osteoblasts in vitro. These data will help in understanding the molecular mechanisms of therapeutic efficacy of sweroside, and highlight insights into drug discovery. In the current study, sweroside has been suggested to be a promising osteoporosis therapeutic natural product.

[Promoting effect of constituents in plasma after oral administration of liuwei dihuangwan on proliferation of rat osteoblast].

OBJECTIVE: To elucidate the promoting effect of the constituents in plasma after oral administration of Liuwei Dihuangwan on proliferation of rat osteoblast. METHOD: To add the constituents in plasma after oral administration of Liuwei Dihuangwan with rational concentration into culture solution of rat osteoblast, later using MT method for determining the proliferation rate of rat osteoblast. RESULT: The Mixed group including morroniside, sweroside and loganin with different dose all significantly promoted proliferation of rat osteoblast. CONCLUSION: This study preliminarily elucidated the constituents such as morroniside, sweroside and loganin in plasma after oral administration of Liuwei Dihuangwan were main biological constituents which contributed to anti-osteoporosis bioactivity of Liuwei Dihuangwan.

[Effects of Naoan tablets on brain hemodynamics and cerebral microcirculation of soft membrane in anesthetized dogs and rats].

OBJECTIVE: To study the effects of Naoan tablets on brain hemodynamics and cerebral microcirculation of soft membrane. METHOD: Cerebral blood stream flux, resistance of blood vessels, blood pressure and heart rate were used as observation indexes in hemodynamics experiment. Artery caliber and the number of capillaries with recovered blood stream were used as observation indexes in microcirculation experiment. RESULT: Naoan tablets at dose of 0.5 g x kg(-1) and 1.0 g x kg(-1) could enhance cerebral blood stream flux, decrease resistance of blood vessels, and reduce blood pressure. While no effects on heart rate. Naoan tablet at dose of 0.7 g x kg(-1) and 2.1 g x kg(-1) could increase the number of capillaries with recovered blood stream and enlarge the artery caliber of soft membrane in rats. CONCLUSION: Naoan tablets can improve the indexes of hemodynamics and cerebral microcirculation of soft membrane.

Analysis and bioactive evaluation of the compounds absorbed into blood after oral administration of the extracts of Vaccinium vitis-idaea in rat.

In order to screen the active constituents of Vaccinium vitis-idaea L., the compounds absorbed into the rat blood after oral administration of ethanol extract of the stems and leaves of V. vitis-idaea (EEV) have been analyzed. Two compounds were detected in the plasma and identified as arbutin and fraxin, which are originally existed in the crude drug. Furthermore, the pharmacological effects of the two compounds involving EEV for curing acute and chronic respiratory tract infection were tested. The results showed that both arbutin and fraxin have the anti-inflammatory, anti-coughing and phlegm-removing effects. Therefore, it was ascertained that the arbutin and fraxin would be the main active constituents of V. vitis-idaea L.