Interactions Between Thrombin and Natural Proudcts of Coreopsis tinctoria Nuttt. and Cistanche deserticola Ma. in Capillary Zone Electrophoresis

Aim A capillary zone electrophoretic method （CZE） was used to determine the interactions between natural products and thrombin. Methods Samples containing natural products and thrombin at various ratios were incubated at 25 ℃ and then were separated by CZE with Tris-acetate buffer at pH 7.2. Each run could be accomplished within 5 min. Results In CZE, the peak width broadened due to the affinity interaction between natural products and thrombin. Compared with positive and negative control, the natural products （CB-1, CB-2） from Coreopsis tinctoria Nuttt. interacted with thrombin; CB-3 from Coreopsis tinctoria Nuttt. and HC-1, HC-2, HC-3 from Cistanche deserticola Ma. did not bind to thrombin. Both qualification and quantification characterizations of the binding were determined. Conclusion The established method is capable of sensitive and fast determination of natural products and thrombin interactions, it can be employed as an alternative method.

Therapeutic effect of Cistanche deserticola on defecation in senile constipation rat model through stem cell factor/C-kit signaling pathway

BACKGROUND Constipation is one of the chronic gastrointestinal functional diseases.It seriously affects the quality of life.Cistanche deserticola(C.deserticola)can treat constipation obviously,but its mechanism has not been clarified.We supposed that mechanism of it improved the intestinal motility by stimulating interstitial Cajal cells(ICC).Activation of the C-kit receptor on the surface of ICC is closely related to ICC function,and the stem cell factor(SCF)/C-kit signaling pathways plays an important role on it.To investigate the mechanism of how C.deserticola treats constipation,this study aimed to establish a constipation model in rats and explore the role of SCF/C-kit signaling pathway in the treatment.AIM To explore the SCF/C-kit signaling pathways in the role of C.deserticola for treatment of constipation by a constipation rat model.METHODS Forty-eight 8-mo-old Sprague–Dawley rats were divided into 4 groups by random weight method:Normal group(n=12),model group(n=12),C.deserticola group(n=12)and blocker group(n=12).The normal group received normal saline by gavage;the model group received loperamide by gavage;the blocker group received loperamide and C.deserticola by gavage,and STI571 was injected by intraperitoneally.During treatment,the weight,fecal granules and fecal quality were recorded every 10 d.On day 20 after model induction,the colon tissues of each group were removed.Hematoxylin and eosin staining was used to observe pathological changes.Expression levels of SCF,C-kit and Aquaporin genes were detected by immunohistochemistry,western blotting,and real-time-quantitative polymerase chain reaction.The colonic epithelial mitochondria and goblet cells were observed by transmission electron microscopy.RESULTS Compared with the normal group,as treatment progressed,the weight of rats in the model and blocker groups decreased significantly,the stool weight decreased,and the stool quality was dry(P<0.05).C.deserticola reversed the decrease in body weight and stool weight and improved stool quality.Histopathological analysis indicated that the colonic mucosal epithelium in the model group was incomplete,and the arrangement of the glands was irregular or damaged.Treatment with C.deserticola improved the integrity and continuity of the epithelial cells and regular arrangement of the glands.The blocking agents inhibited the effects of C.deserticola Immunohistochemistry and real-timequantitative polymerase chain reaction showed that expression of SCF and C-kit protein or genes in the colonic tissue of the model group was decreased(P<0.05),while treatment with C.deserticola increased protein or gene expression(P<0.05).Western blotting showed that expression of aquaporin APQ3 was increased,while the expression of Cx43 decreased in the model group.Treatment with C.deserticola inhibited expression of APQ3 and promoted expression of Cx43.Transmission electron microscopy showed that the mitochondria of the colonic epithelium in the model group were swollen and arranged disorderly,and microvilli were sparse.The condition was better in the C.deserticola group.Mice treated with STI571 blocker confirmed that blocking the SCF/C-kit pathway inhibited the improvement of constipation by C.deserticola.CONCLUSION C.deserticola improved defecation in rats with constipation,and the SCF/C-kit signaling pathway,which is a key link of ICC function,played an important role of the treatment.

Cistanche deserticola decoction alleviates the testicular toxicity induced by hydroxyurea in male mice

This study aimed to evaluate testicular toxicity induced by hydroxyurea （HU） and the possible counteracting effect of an aqueous extract of Cistanche deserticola （CD）. HU is an antineoplastic drug that has potential reproductive toxicity, and Herba Cistanche has been used as a tonic for the reproductive system for thousands of years. Sixty mice were randomly divided into five groups. Except mice in normal group, the rest received HU （400 mg kg^-1 body weight） intragastrically. Meanwhile, mice in normal and HU control groups received purified water, and the rest received intragastrically three doses of CD decoctions （1.5, 3.0 and 6.0 g crude drug kg^-1 body weight, respectively） daily for 4 weeks. Severe testes lesions were observed, testes weight （P〈0.01） and serum luteinising hormone levels （P〈0.0 1） were also decreased significantly, in the HU groups. Three doses of CD decoctions alleviated the spermatogenetic cell degeneration induced by HU and modulated the serum sex hormones levels to some extent.

Folk nomenclature of plants in Cistanche deserticola-associated community in South Gobi, Mongolia

Cistanche deserticola is an important medicinal plant in Mongolia.Despite its significant role in local healing systems,little traditional knowledge had been reported.The present study investigated folk names of C.deserticola and other species of the same community in Umnugobi Province,South Gobi region of Mongolia,based on ethnobotanical approaches.The high correspondence between folk names and scientific names of plant species occurring in Cistanche-associated community shows the scientific meaning of folk nomenclature and classification in Mongolia.The Mongolian and folk names of plants were formed on the basis of observations and understanding of wild plants including their morphology,phenology and traditional uses as well.Results from this study will support the conservation of C.deserticola itself,a rare and endangered plant species listed in the Monglian Red Data Book.Our documentation of folk nomenclature based on 96 plant species in the Cistanche community,as a part of traditional knowledge associated with biodiversity,will be very helpful for making strategy of plant biodiversity conservation in Mongolia.

Cistanche Deserticola for Regulation of Bone Metabolism:Therapeutic Potential and Molecular Mechanisms on Postmenopausal Osteoporosis

Osteoporosis is a generalized disease of bone that leads to a loss of bone density and bone mass,destruction of bone microstructure,increased brittleness and therefore fracture.At present,the main treatment of Western medicine is drug therapy such as bisphosphonates,calcitriol,vitamin D,etc.However,long-term use of these drugs may bring some adverse reactions.Chinese herbal medicine Cistanche deserticola could regulate bone metabolism by promoting osteoblast activity and inhibiting osteoclast activity with low toxicity and adverse reactions.Therefore,Cistanche deserticola has attracted increasing attention for its efficacy in the prevention and treatment of osteoporosis in recent years.Here we present a literature review of the molecular pathways involved in osteoporosis and the effects of Cistanche deserticola on bone metabolism.Our objective is to clarify the mechanism of Cistanche deserticola in the treatment of osteoporosis.

Protective effect of Cistanche deserticola on gentamicin-induced nephrotoxicity in rats

Objective: Gentamicin(GM) is a commonly used aminoglycoside antibiotic, however, renal toxicity has limited its usage. The present study was designed to evaluate the ameliorative effect of Cistanche deserticola on GM-induced nephrotoxicity in rats.Methods: The nephrotoxicity in rats was induced by intraperitoneal administration of GM(100 mg/kg)for 10 consecutive days. Glomerular filtration rate, blood urea nitrogen, creatinine and kidney histopathology were detected to assess the GM-induced nephrotoxicity. The oxidative stress(catalase,superoxide dismutase, glutathione and malondialdehyde) was assessed. The inflammatory response(tumor necrosis factor-a, interleukin-6, myeloperoxidase and nuclear factor-kappa B) and apoptotic marker(Bax and Bcl-2) were also evaluated.Results: The results showed that water and 75% ethanol extracts of C. deserticola(named CDW and CDE,respectively)(100, 200 and 400 mg/kg) in combination with GM could recover the reduction of glomerular filtration rate and enhance the renal endogenous antioxidant capability induced by GM. The increase in the expression of renal inflammatory cytokines(tumor necrosis factor-a and interleukin-6), nuclear protein of nuclear factor-kappa B(p65) and the activity of myeloperoxidase induced by GM was significantly decreased upon CDW or CDE treatment. In addition, CDW or CDE treatment could decrease the Bax protein expression and increase the Bcl-2 protein expression in GM-induced nephrotoxicity in rats significantly.Conclusion: The study demonstrated that C. deserticola treatment could attenuate kidney dysfunction and structural damage in rats induced by GM through the reduction of inflammation, oxidative stress and apoptosis.

Application of ~1H NMR-based metabolomics for discrimination of different parts and development of a new processing workflow for Cistanche deserticola

Cistanche deserticola(CD) is one of the two authoritative source plants of Cistanches Herba, a well-known medicinal plant. Herein,~1H NMR spectroscopy was employed to characterize the chemical profile and to distinguish the different parts, as well as to propose a new processing workflow for CD.Signal assignment was achieved by multiple one and two dimensional NMR spectroscopic techniques in combination with available databases and authentic compounds. The upper parts of the plant were distinguished from the lower parts by combining ~1H NMR spectroscopic dataset with multivariate statistical analysis. A new processing method that hyphenated steaming with freeze-drying, was demonstrated to be superior to either steaming coupled with oven-drying or direct freeze-drying via holistic ~1H NMR-based metabolomic characterization. Phenylethanoid glycosides, mainly echinacoside and acteoside, were screened out and confirmed as the chemical markers responsible for exhibiting the superiority of the new processing workflow, whereas serial primary metabolites, especially carbohydrates and tricarboxylic acid cycle metabolites, were found as the primary molecules governing the discrimination between the upper and lower parts of the plant. Collectively,~1H NMR spectroscopy was demonstrated as a versatile analytical tool to characterize the chemical profile and to guide the indepth exploitation of CD by providing comprehensive qualitative and quantitative information.

Effects of feeding tyrosine and phenylalanine on the accumulation of phenylethanoid glycosides to Cistanche deserticola cell suspension culture

AIM: To investigate the effects of feeding phenylalanine(Phe) and tyrosine(Tyr) on the accumulation of total phenolic compounds and four phenylethanoid glycosides(PeGs) to a cell suspension culture of the parasitic plant Cistanche deserticola. METHOD: A cell suspension culture of C. deserticola was established and precursors of different concentrations were fed. In each group, the cell was sampled at the 24th day after inoculation. The content of total phenolic compounds and four PeGs compounds were determined using the Folin-Ciocalteu method and an HPLC method, respectively. RESULTS: In the Phe fed cells, the maximum PeGs yield was achieved when Phe was fed at 1.5 mmol·L-1 and the yield reached 1.13 times the control cell concentration. In the Tyr fed cells, the maximum yield of PeGs was 1.60 times of control when 0.75 mmol·L-1 Tyr was fed to the cells. Furthermore, it was found that the salidroside yield was 4.01 times of control group when 5 mmol·L-1 Tyr was fed. CONCLUSION: Tyr is a better precursor for PeGs accumulation compared with Phe, and the rate limiting enzymes might be involved in the Tyr branch.

Identification of HPLC Characteristic Chromatograms of Phenylethanoid Glycosides and Iridoid Glycosides in Herba Cistanche of Different Origins

[Objectives]This study aimed to establish a quality evaluation method for Herba Cistanche based on the HPLC characteristic chromatograms of the phenylethanoid glycosides and iridoid glycosides,and compare the HPLC characteristic chromatograms of different origins of Herba Cistanche.[Methods]The chromatographic conditions used were as follows:column,Agilent ZORBAX Eclipse Plus C18 column(4.6 mm×250 mm,5μm);gradient elution,acetonitrile-0.1%phosphoric acid aqueous solution;flow rate,1.0 mL/min;detection wavelength,240 nm;column temperature,30℃.[Results]Cistanche tubulosa(Schenk)Wight showed 9 characteristic peaks,Cistanche deserticola Y.C.Ma showed 10 characteristic peaks,Cistanche salsa(C.A.Mey.)G.Beck showed 7 characteristic peaks,and Cistanche sinensis G.Beck showed 4 characteristic peaks.Peak 6 was echinacoside,peak 7 was verbascoside,peak 8 was tubuloside A,peak 9 was isoacteoside and peak 10 was cistanoside A.The characteristic chromatograms of the four different origins of Herba Cistanche were significantly different.[Conclusions]This characteristic chromatogram method has good reproducibility and can be used to distinguish 4 different origins of Herba Cistanche,C.deserticola Y.C.Ma,C.tubulosa(Schenk)Wight,C.sinensis G.Beck and C.salsa(C.A.Mey.)G.Beck.

Effect of Roucongrong(Herba Cistanches Deserticolae) on reproductive toxicity in mice induced by glycoside of Leigongteng(Radix et Rhizoma Tripterygii)

OBJECTIVE： The purpose of this research is to study the effect of Roucongrong （Herba Cistanches Deserticolae） on reproductive toxicity in mice in- duced by a glycoside extracted from Leigongteng （Radix et Rhizoma Tripterygii） （GRT）. METHODS： Forty-eight BALB/c mice were random-ly divided into two groups in the ratio of I ： 3, 12 in one group and 36 in the other. The 12-mouse group was the control group that was intragastrical- ly administered physiological saline for 3 weeks. The 36 mice in the other group were given 30 mg- kg ·^-1 d i GRT for 3 weeks, then randomly divided into 3 subgroups： the model group, GRI group and Roucongrong （Herba Cistanches Deserticolae） group, with 12 mice in each group. In the model group, 0.25 mL physiological saline was intragastri- cally administered; in the GRT group, GRT, 0.25 mL at 30 mg · kg ^-1· d ^-1 was intragastrically adminis- tered once a day; in the Roucongrong （Herba Cis- tanches Deserticolae） group, mice were adminis- tered Roucongrong （Herba Cistanches Deserticolae） decoction equivalent to 0.25 mL at a final dose of 10g.kg 1.d.1 crude drug （calculated as per 20 times of 0.5 g· kg^-1. d ^-1 for adults）, and GRT 0.25 mL at 30 mg- kg^-1· d^- 1 daily. After another 3 weeks of exposure, expression levels of the reproduction-re- lated genes DEAD （Asp-Glu-Ala-Asp） box polypep- tide 3, Y-linked, B-cell CLL/lymphoma 6 and Signal transducer and activator of transcription 3 were evaluated. RESULTS： After 6 weeks of GRT treatment, the sper- matogenic cell population in the convoluted tu- bule of testis was in disorder and the tubule cavity expanded. Sertoli cell and Leydig cells exhibited at- rophy or disappeared. The number of sperm de- creased. The spermatogenic cell level of testis for male mice was ranked in order and sperm was pro- duced in the cavity of the spermatogenic cell. The expression levels of DDX3Y, BCL6 and STAT3 were up-regulated.CONCLUSION： GRT affected reproduction-related genes. Roucongrong （Herba Cistanches Desertico- lae） reversed reproductive toxicity in mice induced by GRT.