Traditional Uygur Medicine Research Articles

Clerodane diterpenoids from the Uygur medicine Salvia deserta with immunosuppressive activity

Six undescribed clerodane diterpenoids along with five known ones were isolated from the aerial parts of Salvia deserta, a traditional Uygur medicine. Their chemical structures including absolute configurations were elucidated by extensive spectroscopic analysis (including 1D and 2D NMR, HRESIMS, and IR), combined with calculated ECD method and single-crystal X-ray diffraction analysis. All the compounds possessed a terminal α,β-unsaturated-γ-lactone moiety, and were assayed for their immunosuppressive activity via inhibiting the secretion of cytokines TNF-α and IL-6 in macrophages RAW264.7. Among them, (5R,8R,9S,10R)-18-nor-cleroda-2,13-dien-16,15-olide-4-one obviously suppressed the secretion of TNF-α and IL-6 with IC50 values of 8.55 and 13.65 μM, respectively.

Tilianin improves lipid profile and alleviates atherosclerosis in ApoE−/− mice through up-regulation of SREBP2-mediated LDLR expression

BackgroundThe huge global burden of atherosclerotic cardiovascular diseases (CVDs) represents an urgent unmet need for the development of novel therapeutics. Dracocephalum moldavica L. has been used as a traditional Uygur medicine to treat various CVDs for centuries. Tilianin is a major flavonoid component of D. moldavica L. and has potential for preventing atherosclerosis. However, the molecular mechanisms that tilianin attenuate atherosclerosis are far from fully understood. PurposesThe purpose of this study is to investigate the efficiency and underlying mechanisms of tilianin in controlling lipid profile and preventing atherogenesis. MethodsThe lipid-lowering effect of tilianin was evaluated in C57BL/6 and ApoE−/− mice by systematically determining serum biochemical parameters. The effects of tilianin on the atherosclerotic lesion were observed in aortic roots and whole aortas of ApoE−/− mice with oil red O staining. Caecal content from ApoE−/− mice were collected for 16S rRNA gene sequence analysis to assess the structure of the gut microbiota. The inhibition of hepatosteatosis was verified by histological examination, and a liver transcriptome analysis was performed to elucidate the tilianin-induced hepatic transcriptional alterations. Effects of tilianin on the expression and function of LDLR were examined in HepG2 cells and ApoE−/− mice. Further mechanisms underlying the efficacy of tilianin were investigated in HepG2 cells. ResultsTilianin treatment improved lipid profiles in C57BL/6 and dyslipidemic ApoE−/− mice, especially reducing the serum LDL-cholesterol (LDL-C) level. Significant reductions of atherosclerotic lesion area and hepatosteatosis were observed in tilianin-treated ApoE−/− mice. The altered gut microbial composition in tilianin groups was associated with lipid metabolism and atherosclerosis. The liver transcriptome revealed that tilianin regulated the transcription of lipid metabolism-related genes. Then both in vitro and in vivo analyses revealed the potent effect of tilianin to enhance hepatic LDLR expression and its mediated LDL-C uptake. Further studies confirmed a critical role of SREBP2 in hepatic LDLR up-regulation by tilianin via increasing precursor and thus mature nuclear SREBP2 level. ConclusionThis study demonstrated the lipid-lowering effect of tilianin through SREBP2-mediated transcriptional activation of LDLR. Our findings reveal a novel anti-atherosclerotic mechanism of tilianin and underlie its potential clinical use in modulating CVDs with good availability and affordability.

Chemical constituents from the seeds of Nigella glandulifera and their hypoglycemic activities.

The seeds of Nigella glandulifera Freyn et Sint. are traditional Uygur medicine used for the treatment of diabetes. However, the active anti-diabetic constituents in the seeds of N. glandulifera remain unclear. In the present study, a new delabellane-type diterpene, 8-denicotinoylnigellamine A1 (1), and a new acyclic sesquiterpene, 2,6,10-trimethyl-6,7,12-trihydroxy-dodec-2-ene (3), together with eight known compounds including alkaloids (2 and 7), triterpenoid saponins (4–6), and phenolic compounds (8–10), were isolated from the seeds of N. glandulifera. Their structures were determined by extensive spectroscopic analyses and quantum chemical calculations. We evaluated the potential protective effects of the isolated compounds on an insulin resistant HepG2 (IR-HepG2) cell model. The results showed that compounds 2, 4–8, and 10 could promote the consumption of glucose in IR-HepG2 cells. Those compounds might be responsible for the anti-diabetic effects of the seeds of N. glandulifera.

Aloperine Relieves Type 2 Diabetes Mellitus via Enhancing GLUT4 Expression and Translocation.

Aloperine (ALO), a quinolizidine alkaloid isolated from Sophora alopecuroides L. used in the traditional Uygur medicine, induced a significant increase in cellular glucose uptake of L6 cells, suggesting it has the potential to relieve hyperglycemia. Therefore, we investigated the effects of ALO on type 2 diabetes mellitus (T2DM) through in vitro and in vivo studies. The translocation of glucose transporter 4 (GLUT4) and changes in intracellular Ca2+ levels were real-time monitored in L6 cells using a laser scanning confocal microscope and related protein kinase inhibitors were used to explore the mechanism of action of ALO. Furthermore, high fat diet combined with low-dose streptozotocin (STZ) was used to induce T2DM in rats, and ALO was given to the stomach of T2DM rats for 4 weeks. In vitro results showed that ALO-induced enhancement of GLUT4 expression and translocation were mediated by G protein-PLC-PKC and PI3K/Akt pathways and ALO-enhanced intracellular Ca2+ was involved in activating PKC via G protein-PLC-IP3R-Ca2+ pathway, resulting in promoted GLUT4 plasma membrane fusion and subsequent glucose uptake. ALO treatment effectively ameliorated hyperglycemia, glucose intolerance, insulin resistance and dyslipidemia, alleviated hepatic steatosis, protected pancreatic islet function and activated GLUT4 expression in insulin target tissues of T2DM rats. These findings demonstrated that ALO deserves attention as a potential hypoglycemic agent.

Chemical constituents of Anchusa italica Retz. and their protective effects on cardiomyocytes injured by hypoxia/reoxygenation

Abstract Anchusa italica Retz. (family Boraginaceae) is widely distributed in Xinjiang Uygur Autonomous Region, China. Its aerial parts are used as a traditional Uygur medicine for treating cardiovascular and cerebrovascular diseases. Three new triterpenoids (1 − 3), and eleven known compounds were isolated from the aerial parts of A. italica, including six oleanane (1 and 3-7) and eight ursane (2 and 8-14) triterpenoids. Among them, compound 2 was a 19-oxo-1819-seco-ursane-type triterpenoid first isolated from the genus Anchusa. Compounds 2, 6, 7, 10, 11 and 13 exhibited protective effects on neonatal rat cardiomyocytes injured by hypoxia/reoxygenation (H/R). The findings suggest that triterpenoids might be the active constituents of A. italica for treating cardiovascular and cerebrovascular diseases in human, and represent potential compounds for developing cardiomyocyte- protection drugs.

Simultaneous determination of gallic acid, methyl gallate, and 1,3,6-tri-O-galloyl-β-d-glucose from Turkish galls in rat plasma using liquid chromatography-tandem mass spectrometry and its application to pharmacokinetics study.

Turkish galls (TG) is a traditional Uygur medicine typically used in clinics for dental disease and chronic ulcerative colitis. In this study, a novel liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous quantification of gallic acid, methyl gallate, and 1,3,6-tri-O-galloyl-β-d-glucose in rat plasma, which are the major bioactive compounds of TG. After a feasible protein precipitation using acetonitrile for sample preparation, chromatographic separation was performed with a BDS Hypersil C18 column (2.1 × 100 mm, 5 μm) at 30°C, and water containing 10 mmol of ammonium acetate and acetonitrile was used as the mobile phase with a flow rate of 0.3 mL/min. The MS detector was operated in the selective reaction monitoring with negative-ionization mode. The results of the method validation, including selectivity, linearity, accuracy, precision, extraction recovery, matrix effect, and stability of the compounds in the biosamples, were all within the current acceptance criteria. The established method was successfully applied to the pharmacokinetics study of three analytes in rats after an oral administration of TG extract and laid the foundation for studying the active components and mechanism of TG in vivo.

Triterpenoids from Anchusa italica and their protective effects on hypoxia/reoxygenation induced cardiomyocytes injury.

Six new triterpenoids (1-6) and 22 known analogues (7-28), were separated from the aerial parts of Anchusa italica Retz., a traditional Uygur medicine for treating cardiovascular and cerebrovascular diseases in the Xinjiang region, China. The possible effects of compounds 1-28 on hypoxia/reoxygenation (H/R) induced cardiomyocytes injury were assayed, and compounds 4, 6-17, 21-22 and 26-28 showed significant protective effects. Further, the representative new compound 6 significantly suppressed the levels of H/R-induced apoptosis and autophagy in neonatal rat cardiomyocytes, with the reversing of the downregulated expression of Bcl-2 and upregulated expression of Bax and Beclin-1 by compound 6 treatment in neonatal rat cardiomyocytes following H/R injury. In addition, compound 6 protected cardiomyocyte from H/R injury, and pretreatment with 6 could decrease CK and LDH levels. Compound 6 also alleviated H/R-induced phosphorylation of p38 MAPK in neonatal rat cardiomyocytes. Therefore, tripterpenoid 6 and its analogues may be the pharmacodyamic material of A. italica, and offer a promising therapeutic approach for treating cardiomyocyte injury induced by H/R.

Simultaneous determination of five N-alkylamides in the root of Anacyclus pyrethrum by HPLC and profiling of components in its methanolic root extract by UPLC/Q-TOF-MS

The root of Anacyclus pyrethrum (L.) Lag., Asteraceae, is very widely used for treating various diseases in Traditional Uygur Medicine, particularly in the treatment of vitiligo. However, there have been few studies on the quality standards of A. pyrethrum in China. A. pyrethrum contains abundant N-alkylamides, which are considered to be the principal components. Therefore, based on the previous research in our group, six N-alkylamides were obtained by using column chromatography. We used ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry to determine the mass spectrometry cleavage mechanism of these six monomer components and established the mass spectrometry cleavage law of N-alkylamides. Then, we used the ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry method to rapidly identify and analyze the N-alkylamide components of the A. pyrethrum methanol extract. Finally, twenty N-alkylamides were identified, including eleven N-isobutylamides, two N-methyl isobutylamides, six 4-hydroxyphenylethyl-amide and one 2-phenylethylamide. Five of these compounds were identified as new compounds that have not been reported to date. Two of these compounds were identified for the first time in this herb. Therefore, this work provides an approach for the quality analysis of N-alkylamides in the root of A. pyrethrum. A search of the literature showed that the content determination in the A. pyrethrum quality standard is still a remaining problem. N-alkylamides are the main components of A. pyrethrum. Even though ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry has the advantages of lower time and higher efficiency compared to high-performance liquid chromatography, considering the ease of repeatability and universality of the quality control method, we chose to use high-performance liquid chromatography for content determination. In this experiment, high-performance liquid chromatography was used for the first time to establish a simple, rapid and accurate method for evaluating the N-alkylamide content in A. pyrethrum with five N-alkylamides used as the standards. Finally, this work provides a qualitative and quantitative method for the analysis of N-alkylamides in A. pyrethrum, improving the quality control standards for A. pyrethrum.

Determination and pharmacokinetic study of guaiol in rat plasma by gas chromatography–mass spectrometry with selected ion monitoring

Guaiol has been used for thousands of years as a traditional Uygur medicine and is the primary active component found in Ferula ferulaeoides (Steud.) Korov (F. ferulaeoides). In our present study, a rapid, selective, and sensitive method of monitoring selected ions was established based on gas chromatography-mass spectrometry. This method was optimized for the quantification and pharmacokinetic analysis of guaiol in rat plasma following oral administration of chloroform extract from Ferula ferulaeoides. Plasma was extracted using liquid-liquid extraction with ethyl acetate and was analyzed on a HP-5MS column (30 m × 250 μm × 0.25 μm) with a mass selective detector. Detection was carried out under selected ion monitoring mode, and three selected ion monitoring ions (m/z 59.1, 107.1, and 161.1 for guaiol) were used for the quantitative determination of that under investigation. The assay demonstrated excellent linearity in the range of 1–200 ng/mL (r = 0.9993, n = 8) in the case of guaiol measured in rat plasma. The limit of detection and the limit of quantification for guaiol in rat plasma were found to be 0.25 ng/mL and 1 ng/mL, respectively. Intra-day and inter-day precisions were expressed as the relative standard deviation for the method and were in the range of 97.49%–106.16% and 97.04%–105.91%, respectively. Extraction efficiencies were all determined to be >90%, and recoveries were ranged from 91.25% to 96.24%. This method has been successfully applied for the pharmacokinetic evaluation of chloroform extract isolated from F. ferulaeoides following a single oral administration dose (157.5 mg/kg) in rats. The guaiol pharmacokinetic study demonstrated that the half-life of guaiol was 9.18 ± 3.75 h, the mean residence time was 9.07 ± 3.86 h, the maximum guaiol concentration in the plasma was 28.63 ± 6.82 ng/mL, and the maximum time guaiol was in the plasma was 0.50 h.

Evaluation on Analgesic and Anti-Inflammatory Activities of Total Flavonoids from Juniperus sabina.

The leaves of Juniperus sabina (Cupressaceae) are used in traditional Uygur medicine for the treatment of rheumatism and arthritic pain. This study aimed to investigate the analgesic and anti-inflammatory effects of total flavonoids from leaves of Juniperus sabina (JSTF) on rodents. The anti-inflammatory activity was investigated using the carrageenan, egg albumin, or histamine-induced rat paw edema as well as xylene-induced ear edema, capillary permeability, and cotton pellet granuloma while the antinociceptive activity was evaluated using the mouse writhing, formalin, and hot-plate tests. JSTF (125, 250, 500 mg/kg) significantly inhibited xylene-induced ear edema in mice (inhibition ratio as 16.22%, 40.67%, and 51.78%, respectively) and also significantly ameliorated acetic acid increased vascular permeability in mice (inhibition ratio as 11.63%, 32.56%, and 53.49%, respectively). JSTF (250 and 500 mg/kg) gave significant reduction of carrageenin-induced paw oedema at the interval of 1 h and 5 h. Administration of JSTF (500 mg/kg) caused a significant anti-inflammatory effect against oedema induced by egg albumin or histamine at the interval of 0.5 h and 4 h, and both which induced the paw oedema were also inhibited by JSTF (250 mg/kg) at a point in 1, 2, or 3 h after the inflammation. Furthermore, JSTF (125, 250, and 500 mg/kg) produced time-course increase in pain threshold in hot-plate test also. JSTF produced concentration-dependent inhibition on cyclooxygenase-2 (COX-2) or 5-lipoxygenase (5-LO) activities in vitro, and their IC50 values were 31.92 and 129.26 μg/mL, respectively. Moreover, JSTF significantly caused a significant dose-dependent inhibition on acetic acid induced writhing response in mice (inhibition ratio as 23.27%, 36.91%, and 50.76%, respectively). JSTF also significantly ameliorated formalin-induced pain in mice in the late phase on dose-dependent way. These results confirms the clinical use of J. sabina for treatment of rheumatoid arthritis in ethnomedicine, and its effective mechanism will be further studied in the future.

A Systems Pharmacology Approach to Determine Active Compounds and Action Mechanisms of Xipayi KuiJie\u2019an enema for Treatment of Ulcerative colitis

Xipayi Kui Jie’an (KJA), a type of traditional Uygur medicine (TUM), has shown promising therapeutic effects in Ulcerative colitis (UC). Owing to the complexity of TUM, the pharmacological mechanism of KJA remains vague. Therefore, the identification of complex molecular mechanisms is a major challenge and a new method is urgently needed to address this problem. In this study, we established a feasible pharmacological model based on systems pharmacology to identify potential compounds and targets. We also applied compound-target and target-diseases network analysis to evaluate the action mechanisms. According to the predicted results, 12 active compounds were selected and these compounds were also identified by HPLC-ESI-MS/MS analysis. The main components were tannins, this result is consistent with the prediction. The active compounds interacted with 22 targets. Two targets including PTGS2 and PPARG were demonstrated to be the main targets associated with UC. Systematic analysis of the constructed networks revealed that these targets were mainly involved in NF-κB signaling pathway. Furthermore, KJA could also regulate the CD4 + CD25 + Foxp3 + Treg cells. In conclusion, this systems pharmacology-based approach not only explained that KJA could alleviate the UC by regulating its candidate targets, but also gave new insights into the potential novel therapeutic strategies for UC.

Hypoglycemic effect of the polyphenols rich extract from Rose rugosa Thunb on high fat diet and STZ induced diabetic rats

Ethnopharmacological relevanceRosa rugosa Thunb is a traditional Uygur medicine that has been used in the treatment of diabetes in Uygur ancient recipe for hundreds of years. However, the mechanism of Rosa rugosa Thunb activity is still unclear. This study was designed to address this issue by studying the polyphenols enriched extract (RPE) of Rosa rugosa Thunb in diabetic rats. Materials and methodsRPE were tested in the inhibition of α-glucosidase and oxidative stress in vitro. RPE was administrated at dosages of 37.5, 75 and 150mg/kg body weight in the type 2 diabetic rats, which were made by high fat diet feeding plus a low dose of STZ injection (30mg/kg). The therapeutic effect was evaluated four weeks later. Oral glucose tolerance test (OGTT), insulin tolerance test (ITT) and insulin signal pathway (PI3K/AKT) were examined to determine insulin sensitivity. Blood glucose levels and body weight were measured weekly in the study. ResultsIn vitro, RPE exhibited an activity in the inhibition of α-glucosidase and had an excellent antioxidant activity in the liver of diabetic rats. RPE significantly decreased the fasting blood glucose, improved insulin sensitivity (HOMA-IR), OGTT, ITT and blood lipid profile. The glycogen synthesis and hexokinase activity were increased together with the improved signaling activity of insulin as indicated by p-IRS, p-IR, p-AKT, and p-GSK-3β. ConclusionsThe results suggest that RPE reduced blood glucose in type 2 diabetic rats by improvement of insulin sensitivity. The effect is likely achieved by inhibition of oxidative stress and α-glucosidase.

The treatment of Uygur medicine Dracocephalum moldavica L on chronic mountain sickness rat model.

Aim:Dracocephalum moldavica L, a traditional Uygur medicine, possesses some key cardiac activities. However, till date, no reports are available on the use of D. moldavica against chronic mountain sickness (CMS), which is a medical condition that affects the residents of high altitude. The present study was designed to explore the treatment efficacy of D. moldavica on CMS.Materials and Methods:80 of the 100 Sprague Dawley rats enrolled were bred in simulated high altitude environment and the remaining 20 rats were kept in the plains. Water and alcohol extracts of D. moldavica were prepared. CMS rat model was prepared, and the rat hearts were removed for histopathological analysis. Blood samples were taken for hematological and biochemical analyses. Rat pulmonary artery pressure was determined to study the treatment efficacy.Results:In the CMS model group, the levels of interleukin-6 (IL-6), C-reactive protein (CRP), and malondialdehyde (MDA) were found to be significantly higher than the control group; while the concentrations of SOD and GSH-Px decreased. D. moldavica could improve these levels, decrease pulmonary artery pressure, and improve the cardiac pathological state.Conclusions:The study results show that IL-6, CRP, MDA, SOD and GSH-Px participate and mediate the formation of CMS and D. moldavica is found to possess noticeable effects on CMS. The present study explored the basics of high altitude sickness and laid the foundation for further progress of Uygur medicines on the treatment of altitude sickness. Further preclinical and clinical studies with more sample size are recommended.

Development of a rapid resolution liquid chromatography-diode array detector method for the determination of three compounds in Ziziphora clinopodioides Lam from different origins of Xinjiang

Context:As a traditional Uygur medicinal plant, Z. clinopodioides Lam has various uses in Xinjiang.Aims:A reversed-phase rapid resolution liquid chromatography (RP-RRLC) method with diode array detector (DAD) was developed for simultaneous determination of diosmin, linarin, and pulegone from Ziziphora clinopodioides Lam, a widely used in traditional Uygur medicine for treating heart disease, high blood pressure, and other diseases.Settings and Design:Compounds were separated on a XDB-C18 reversed-phase analytical column (50 mm × 4.6 mm, 1.8 μm) with gradient elution using methanol and 1% aqueous acetic acid (v/v) at 0.9 mL/min. he detection wavelength was set at 270 nm.Materials and Methods:Ziziphora clinopodioides Lam. were collected from ten different origins in Xinjiang, including the Ban fang ditch, Tuoli, the Altay mountains, Terks, Xiata Road, Zhaosu Highway, Guozigou, Fukang, Jimsar, Wulabo.Statistical Analysis Used:The intra-day and inter-day precisions of all three compounds were less than 0.89% and the average recoveries ranged from 97.4 to 104.1%. There were highly significant linear correlations between component concentrations and specific chromatographic peak areas (R2 > 0.999).Results:The proposed method was successfully applied to determine the levels of three active components in Z. clinopodioides Lam. samples from different locations in Xinjiang.Conclusions:The proposed method is simple, consistent, accurate, and could be utilized as a quality control method for Z. clinopodioides Lam.

Chemical composition of the essential oil of Hyssopus cuspidatus from Xinjiang, China

The genus Hyssopus (Labiatae), consisting of about 15 species, is distributed in Central Asia, South Europe, and North Africa. In China, there are only two species, Hyssopus cuspidatus Boiss and H. latilabiatus C. Y. Wu et H. w. Li, all growing in Xinjiang [1]. Both of them have been used as nectar source plants and medicinal plants. In Xinjiang, H. cuspidatus Boriss. has also been used in traditional Uygur medicine in the treatment of cough, asthma, bronchitis, trauma, and rheumatism [2, 3]. As the major Hyssopus resource for medical use in China, there are few reports on the essential oil of Hyssopus cuspidatus previously. We identified a total of 36 compounds in the hydrodistilled essential oil of Hyssopus cuspidatus from Xinjiang, China, with germacrene D (18.67%), hexadecanoic acid (17.53%), germacrene B (15.61%), (+)trans-caryophyllene (8.04%), and (+)spathulenol (4.11%) as the main components (Table 1). Further investigation of Hyssopus cuspidatus essential oil is still needed.