Xingbei antitussive granules ameliorate cough hypersensitivity in post-infectious cough guinea pigs by regulating tryptase/PAR2/TRPV1 pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Xingbei antitussive granules (XB) is a classic Chinese Medicine prescription for treating post-infectious cough(PIC), based on the Sanao Decoction from Formularies of the Bureau of People's Welfare Pharmacies in the Song Dynasty and Jiegeng decoction from Essentials of the Golden Chamber in the Han Dynasty. However, the therapeutic effects and pharmacological mechanisms are still ambiguous. In the present study, we endeavored to elucidate these underlying mechanisms. AIMS OF THE STUDY: This study aimed to explore the potential impact and mechanism of XB on PIC, and provide a scientific basis for its clinical application. MATERIALS AND METHODS: Cigarette smoking (CS) combined with lipopolysaccharide (LPS) nasal drops were administered to induce the PIC guinea pig with cough hypersensitivity status. Subsequently, the model guinea pigs were treated with XB and the cough frequency was observed by the capsaicin cough provocation test. The pathological changes of lung tissue were assessed by HE staining, and the levels of inflammatory mediators, mast cell degranulating substances, and neuropeptides were detected. The protein and mRNA expression of transient receptor potential vanilloid type 1(TRPV1), proteinase-activated receptor2(PAR2), and protein kinase C (PKC) were measured by Immunohistochemical staining, Western blot, and RT-qPCR. Changes in the abundance and composition of respiratory bacterial microbiota were determined by 16S rRNA sequencing. RESULTS: After XB treatment, the model guinea pigs showed a dose-dependent decrease in cough frequency, along with a significant alleviation in inflammatory infiltration of lung tissue and a reduction in inflammatory mediators. In addition, XB high-dose treatment significantly decreased the levels of mast cell Tryptase as well as ß-hexosaminidase (ß-Hex) and downregulated the expression of TRPV1, PAR2, and p-PKC. Simultaneously, levels of neuropeptides like substance P (SP), calcitonin gene-related peptide (CGRP), neurokinin A (NKA), and nerve growth factor (NGF) were improved. Besides, XB also can modulate the structure of respiratory bacterial microbiota and restore homeostasis. CONCLUSION: XB treatment alleviates cough hypersensitivity and inflammatory responses, inhibits the degranulation of mast cells, and ameliorates neurogenic inflammation in PIC guinea pigs whose mechanism may be associated with the inhibition of Tryptase/PAR2/PKC/TRPV1 and the recovery of respiratory bacterial microbiota.

Didymin alleviates metabolic dysfunction-associated fatty liver disease (MAFLD) via the stimulation of Sirt1-mediated lipophagy and mitochondrial biogenesis.

BACKGROUND: Metabolic dysfunction-associated fatty liver disease (MAFLD) is one of the most prevalent metabolic syndromes worldwide. However, no approved pharmacological treatments are available for MAFLD. Chenpi, one kind of dried peel of citrus fruits, has traditionally been utilized as a medicinal herb for liver diseases. Didymin is a newly identified oral bioactive dietary flavonoid glycoside derived from Chenpi. In this study, we investigated the therapeutic potential of Didymin as an anti-MAFLD drug and elucidated its underlying mechanisms. METHODS: High-fat diet (HFD)-induced MAFLD mice and alpha mouse liver 12 (AML12) cells were utilized to evaluate the effects and mechanisms of Didymin in the treatment of MAFLD. Liver weight, serum biochemical parameters, and liver morphology were examined to demonstrate the therapeutic efficacy of Didymin in MAFLD treatment. RNA-seq analysis was performed to identify potential pathways that could be affected by Didymin. The impact of Didymin on Sirt1 was corroborated through western blot, molecular docking analysis, microscale thermophoresis (MST), and deacetylase activity assay. Then, a Sirt1 inhibitor (EX-527) was utilized to confirm that Didymin alleviates MAFLD via Sirt1. Western blot and additional assays were used to investigate the underlying mechanisms. RESULTS: Our results suggested that Didymin may possess therapeutic potential against MAFLD in vitro and in vivo. By promoting Sirt1 expression as well as directly binding to and activating Sirt1, Didymin triggers downstream pathways that enhance mitochondrial biogenesis and function while reducing apoptosis and enhancing lipophagy. CONCLUSIONS: These suggest that Didymin could be a promising medication for MAFLD treatment. Furthermore, its therapeutic effects are mediated by Sirt1.

Weekly phototherapy is an effective therapy for patients with vitiligo.

The current strategies for the treatment of vitiligo using phototherapy usually involve treatment for two-three times per week; however, in practice, the number of patient sessions does not meet this standard. The present study found that phototherapy once a week was also effective. The present study was designed to examine the efficacy of weekly light therapy. For this purpose, 296 patients with vitiligo were included and divided into five sub-samples of the neck, face, trunk, extremities and scalp according to the site of phototherapy, and were treated once or twice weekly with phototherapy. The difference in efficacy between phototherapy performed once and twice weekly was observed using a Chi-squared test. It was concluded that there was a minimal difference between phototherapy performed twice weekly compared to once weekly for the treatment of vitiligo on the face, neck, torso, limbs and scalp. Thus, phototherapy once a week is valid for the treatment of vitiligo, although weekly light therapy takes longer to restore color for the first time.

Black phosphorus quantum dots induced ferroptosis in lung cell via increasing lipid peroxidation and iron accumulation.

Black Phosphorus Quantum Dots (BP-QDs) have potential applications in biomedicine. BP-QDs may enter the body through the respiratory tract during grinding and crushing production and processing, causing respiratory toxicity. Ferroptosis is an oxidative, iron-dependent form of cell death. Here, respiratory toxicity of BP-QDs has been validated in mice and human bronchial epithelial cells. After 24 h of exposure to different doses (4-32 µg/mL) of BP-QDs, intracellular lipid peroxidation and iron overload occurred in Beas-2B cells. After 4 times exposures by noninvasive tracheal instillation at four doses [0, 0.25, 0.5 and 1 (mg/kg/48h)], all animals were sacrificed, organs were removed, processed for pathological examination and molecular analysis. Iron overload, glutathione (GSH) depletion and lipid peroxidation in the lung tissue of mice in the exposure group. Furthermore, based on the ferroptosis-associated protein and mRNA expression, it was hypothesized that BP-QDs induced ferroptosis through increasing intracellular free iron and polyunsaturated fatty acid synthesis. By comparing with previous studies, we speculate that primary cells generally are more sensitive to BP-QDs-induced damage than cancer cells. In summary, findings in the present study confirmed that BP-QDs induce ferroptosis via increasing lipid peroxidation and iron accumulation in vitro and in vivo.

Eugenol improves salt tolerance via enhancing antioxidant capacity and regulating ionic balance in tobacco seedlings.

Salt stress inhibits plant growth by disturbing plant intrinsic physiology. The application of exogenous plant growth regulators to improve the plant tolerance against salt stress has become one of the promising approaches to promote plant growth in saline environment. Eugenol (4-allyl-2- methoxyphenol) is the main ingredient in clove oil and it is known for its strong antioxidant and anti-microbial activities. Eugenol also has the ability of inhibiting several plant pathogens, implying the potential use of eugenol as an environmental friendly agrichemical. However, little is known about the possible role of eugenol in the regulation of plant tolerance against abiotic stress. Therefore, here we investigated the effectiveness of phytochemical eugenol in promoting salt tolerance in tobacco seedlings through physiological, histochemical, and biochemical method. The seedling roots were exposed to NaCl solution in the presence or absence of eugenol. Salt stress inhibited seedling growth, but eugenol supplementation effectively attenuated its effects in a dose-dependent manner, with an optimal effect at 20 µM. ROS (reactive oxygen species) accumulation was found in seedlings upon salt stress which was further resulted in the amelioration of lipid peroxidation, loss of membrane integrity, and cell death in salt-treated seedlings. Addition of eugenol highly suppressed ROS accumulation and reduced lipid peroxidation generation. Both enzymatic and non-enzymatic antioxidative systems were activated by eugenol treatment. AsA/DHA and GSH/GSSG were also enhanced upon eugenol treatment, which helped maintain redox homeostasis upon salinity. Eugenol treatment resulted in an increase in the content of osmoprotectants (e.g. proline, soluble sugar and starch) in salt-treated seedlings. Na+ levels decreased significantly in seedlings upon eugenol exposure. This may result from the upregulation of the expression of two ionic transporter genes, SOS1 (salt-hypersensitive 1) and NHX1 (Na+/H+ anti-transporter 1). Hierarchical cluster combined correlation analysis uncovered that eugenol induced salt tolerance was mediated by redox homeostasis and maintaining ionic balance in tobacco seedlings. This work reveals that eugenol plays a crucial role in regulating plant resistant physiology. This may extend its biological function as a novel biostimulant and opens up new possibilities for improving crop productivity in the saline agricultural environment.

Protection of Vitamin C on Oxidative Damage Caused by Long-Term Excess Iodine Exposure in Wistar Rats.

Vitamin C was reported to be able to protect against oxidative damage due to its reducibility. 120 Wistar rats were randomly divided into 4 × 2 groups, including normal iodine (NI), high iodine (HI), low vitamin C (HI + LC), and high vitamin C (HI + HC); potassium iodide (KI) and potassium iodate (KIO3) were commonly used as additives for iodized salt, so every group was also divided into KI and KIO3 groups. After 6 months' feed, the activities of antioxidant enzymes and Lipid Peroxide (MDA) content in serum, liver, kidney, brain, thyroid and lens were determined. In serum, for males, long-term excess iodine intake caused oxidative damage; in the liver, male rats in the HI + LC group had the highest MDA content, which showed that low-dose vitamin C might promote oxidative damage; in kidneys, the MDA content in the HI and HI + LC groups of females was higher; in the brain, high-dose vitamin C could increase the activity of superoxide dismutase (SOD), which was decreased by high iodine intake, and it also decreased MDA content; in the thyroid, for KIO3, the activity of SOD in the HI group was lower than NI and HI + LC; in the lens, the MDA content in females was lower than males. Long-term excess iodine exposure caused oxidative damage and showed sex difference, and vitamin C had a protective effect on it, especially for high-dose vitamin C.

In vitro and in silico anti-osteoporosis activities and underlying mechanisms of a fructan, ABW90-1, from Achyranthes bidentate.

Achyranthes bidentata is a traditional Chinese medicine used to treat osteoporosis. AB90, a crude saccharide from A. bidentata, showed excellent osteoprotective effects in ovariectomized rats, and ABW90-1, an oligosaccharide purified from AB90, stimulated significant differentiation of osteoblasts. However, the osteogenic effects and underlying mechanisms of ABW90-1 have remained unknown. In the present study, we found that ABW90-1 significantly promoted ALP activity, mineralization, and the expression of osteogenic markers in MC3T3-E1 cells. ABW90-1 showed strong binding with the WNT signaling complex and BMP2 based on number of interactions, hydrogen bond length, and binding energy in silico. ABW90-1 significantly increased the expression of active-ß-catenin, p-GSK-3ß, LEF-1, BMP2, and p-SMAD1. Importantly, the osteogenic effects of ABW90-1 were partially suppressed by DKK-1 and Noggin, which are specific inhibitors of the WNT and BMP signaling pathways, respectively. Collectively, these findings suggest that ABW90-1 has osteogenic effects through crosstalk between WNT/ß-catenin and BMP2/SMAD1 signaling pathways.

[Analysis on formulation regularity and characteristics of memory-boosting Chinese medicinal health products and Chinese patent medicines].

The present study analyzed the current Chinese medicinal health products and Chinese patent medicines effective in boosting memory,aiming at providing references for the formulation and development of memory-boosting health products. The information on memory-boosting health products published by the Department of Special Food Safety Supervision and Management,the State Administration for Market Regulation( SAMR) was collected and the Chinese patent medicines on DRUGDATAEXPY were searched. Microsoft Excel and the TCMISS were used to statistically analyze the characteristics of formulations. A total of 212 memory-boosting health products were obtained from SAMR,including 83 ones containing Chinese medicinal materials. Twelve Chinese herbal medicines showed a usage frequency ≥ 8,with 151 times in use. In DRUGDATAEXPY,258 similar Chinese patent medicines were collected.Twelve Chinese herbal medicines showed a usage frequency ≥ 58,with 907 times in use. Through unsupervised hierarchical entropybased clustering of the above-mentioned Chinese medicinal health products and Chinese patent medicines separately,5 and 12 new formulas were obtained. The selection of Chinese herbal medicines for the new formulas was consistent with the principles of traditional Chinese medicine( TCM) theories,i. e.,tonifying kidney and marrow,benefiting Qi,nourishing Yin,resolving phlegm,and eliminating stasis. According to TCM theories,syndrome differentiation of the users was conducted,and the formulas were designed following the correspondence of syndromes with formulas and Chinese herbal medicines. This study is expected to provide new ideas and methods for the development of Chinese medicinal health products and accurately guide practical applications to exert the advantages of TCM in health care based on syndrome differentiation and improve the effect of Chinese medicinal health products.

[Analysis on formulation regularity and characteristics of acne-relieving Chinese medicinal health products and Chinese patent medicines].

This study summarized and analyzed the current acne-relieving Chinese medicinal health products and Chinese patent medicines,aiming at providing references for the formulation and development of acne-relieving health products. The information on acne-relieving health products published by the Department of Special Food Safety Supervision and Management,the State Administration for Market Regulation( SAMR) was retrieved,and the Chinese patent medicines on DRUGDATAEXPY were searched. Microsoft Excel and the Traditional Chinese Medicine Inheritance Support System V2. 5( TCMISS) were employed to statistically analyze the characteristics of formulations. Forty-three acne-relieving health products were obtained,including 40 ones containing Chinese herbal medicines. Six Chinese herbal medicines showed a usage frequency ≥8,with 61 times in use totally,including Salviae Miltiorrhizae Radix et Rhizoma( 13),Lonicerae Japonicae Flos( 12),Taraxaci Herba( 11),ALOE( 9),Carthami Flos( 8),and Rhei Radix et Rhizoma( 8). A total of 29 Chinese patent medicines with similar efficacy were collected in DRUGDATAEXPY. Nine Chinese herbal medicines showed a usage frequency ≥6,with 63 times in use,including Rhei Radix et Rhizoma( 10),Glycyrrhizae Radix et Rhizoma( 8),Scutellariae Radix( 8),Lonicerae Japonicae Flos( 7),Angelicae Sinensis Radix( 6),Paeoniae Radix Rubra( 6),Astragali Radix( 6),Paeoniae Radix Alba( 6),and Gardeniae Fructus( 6). Through unsupervised hierarchical entropy-based clustering of the above-mentioned Chinese medicinal health products and Chinese patent medicines,five and three new formulas were obtained,respectively. The selection of Chinese herbal medicines was consistent with the principles of traditional Chinese medicine( TCM) theories in relieving acne,i. e.,dispelling exterior wind,ventilating lungs,clearing heat,dissipating mass,promoting diuresis,relaxing bowels,removing toxin,activating blood,resolving stasis,eliminating phlegm,and regulating Qi. According to the " lung governing skin and body hair" and " interior-exterior relationship between lung and large intestine" in TCM theories,the idea that acne could be treated by relieving constipation was proposed,which provided new methods and references for the research and development of Chinese medicinal health products.

[Literature research of Passiflora incarnata and discussion of its traditional Chinese medicine properties].

Based on the research literatures of Passiflora incarnata and the theory of traditional Chinese medicine, the paper discussed the traditional Chinese medicinal properties of P. incarnate, so as to provide a theoretical basis for the compatibility and application of P. incarnata. The literature databases of CNKI, Wanfang, VIP, Web of Science, PubMed and Scopus were selected, and the literatures relating to P. incarnata were reviewed to screen out the scientific research literatures with a high credibility, rational design and reliable conclusions. Foreign pharmacopoeia was consulted, and the listed products were summarized. The traditional Chinese medicine properties of P. incarnata were studied based on 32 clinical trials, 66 pharmacological researches, 64 chemical constituents researches as well as the theory of traditional Chinese medicine. It was preliminarily concluded that the medicinal properties of P. incarnata are sweet, cool, and enter heart, liver channels. The function is mainly to calm the heart and tranquilizing the mind, and calm the liver wind. It is used for hyperactivity of liver-Yang, stagnation of liver-Qi, restlessness of mind, depression, nervousness, insomnia. This paper summarized the source, characteristics of natures, tastes and channel tropism, usage and dosage, function indications of P. incarnata, and defined its clear traditional Chinese medicine property, which lays a theoretical foundation for the compatibility and clinical application of P. incarnata and Chinese medicine.

[Literature research and traditional Chinese medicine properties of Aspalathus linearis].

Aspalathus linearis is a needle-shaped shrub that grows in the Cedarberg mountains in southern South Africa, with an extremely high medicinal value. In 2014, China has approved A. linearis as a new food material. Through retrieval in CNKI, Wanfang, VIP, Web of Science, PubMed and Scopus databases, the literatures were excluded, classified and summarized.On the basis of Chinese medicine theory, the traditional Chinese medicine properties were deducted. Finally, 264 relevant li-teratures were included and classified into 6 categories: review, planting, chemical composition, clinical study, pharmacological effects and safety. The traditional Chinese medicinal properties were deducted as sweet flavor and neutral property. It enters kidney, spleen, heart and liver channels. The major functions are to tonify the kidney and benefit the essence, nourish Qi and spleen, nourish Yin and prompt the production of body fluid, tranquilize mind, and relieve pain. It can be used for soreness of the waist and fatigue, sexual disinterest, limbs heaviness, thirst due to insufficiency of fluid and internal heat, irritability and insomnia, forget fulness, stomachache, joint pain, dysmenorrhea, headache. Preparation for external use can treat eczema itching. Water decoction(2-15 g) can also be used as tea directly. This paper defined the traditional Chinese medicine properties of A. linearis, so as to provide the theoretical basis for further clinical application.

[Therapeutic effect of Wei's triple nine needling combined with medication on optic atrophy].

OBJECTIVE: To explore the effect of Wei 's triple nine needling on visual acuity and visual field in patients with optic atrophy. METHODS: A total of 90 patients with optic atrophy were randomized into an observation group and a control group, 45 cases in each one. Treatment of Wei 's triple nine needling combined with conventional medication were adopted in the observation group, conventional medication was given in the control group. Treatment for 4 weeks was required in both groups. Before treatment and 2, 4 weeks into treatment, the visual acuity and visual field were observed, and the clinical efficacy was evaluated in both groups. RESULTS: The total effective rate was 57.8% (26/45) in the observation group, which was superior to 28.9% (13/45) in the control group (P<0.05). After 2-week and 4-week treatment, the visual acuity was improved (P<0.01), the mean defect (MD) of visual field was decreased (P<0.01), the mean sensitivity (MS) of visual field was increased in the observation group (P<0.05, P<0.01). After 2-week and 4-week treatment, the visual acuity and the MD of visual field were improved (P<0.01, P<0.05), while the difference of MS of visual field compared before treatment had no statistical significance in the control group (P>0.05). The improvement of visual acuity, MD and MS of visual field after 2-week and 4-week into treatment in the observation group were superior to those in the control group (P<0.05, P<0.01). CONCLUSION: Wei 's triple nine needling can effectively improve the visual acuity and the defect of visual field in patients with optic atrophy.

Genomic Sequence Resource of Alternaria alternata Strain B3 Causing Leaf Blight on Ophiopogon japonicus.

Black spot of Ophiopogon japonicus is an economically important disease, which can result in significant losses of both yield and quality of this traditional Chinese medicinal plant. The disease is caused by the small-spored fungal pathogen Alternaria alternata, a necrotrophic fungus that is ubiquitously distributed in the environment. Here, we present the draft whole-genome sequence of A. alternata strain B3 that caused black spot on O. japonicus. The assembly consists of 76 contigs with an estimated genome size of 33.8 Mb. Furthermore, we identified genes that may be associated with the pathogenicity, such as carbohydrate-active enzymes, secreted proteins, and secondary metabolite gene clusters. This genome resource will provide a useful source for future research on the evolution of pathogenicity of A. alternata and phylogenomic analysis in delineating phylogenetic lineages within the genus Alternaria.

Use of life cycle assessment and water quality analysis to evaluate the environmental impacts of the bioremediation of polluted water.

The lakes along the Yangtze River are important source of pollutants that ultimately flow from the river into the East China Sea. Bioremediation is a green technology used to treat polluted water in lakes along the Yangtze River. Life cycle assessment and a comprehensive water quality index are used to evaluate the potential environmental impacts of constructed wetlands (CWs), ecological floating beds (EFBs), and combined ecological floating beds (CEFBs). The results showed that the raw material acquisition, construction, and operation of the CWs, EFBs, and CEFBs accounted for 24.1%, 35.3%, and 40.6%, respectively, of the total environmental impact. The acquisition of raw materials to construct the bioremediation system accounted for 51.6% of the total environmental impact. Among the nine impact categories considered, the system's global warming potential was the largest. Among the three stages of the project (raw material acquisition, construction, and operation), construction had the largest impact on eutrophication (the eutrophic potential of the construction stage was the largest). Furthermore, the operation of the project reduced the human eco-toxicity potential. The evaluation of the water quality before and after implementing the project revealed that CEFBs purified the water more effectively than CWs and EFBs did, particularly with respect to the removal of the total phosphorus.

[Chemical composition analysis of Eucalyptus essential oil and allelopathic effects of α-terpineol]. / æ¡‰æ ‘æ²¹åŒ–å­¦æˆåˆ†åˆ†æžåŠα-松油醇的化感作用.

We extracted Eucalyptus essential oil by steam distillation and analyzed its chemical components by GC-MS. The inhibitory effects of α-terpineol, a component in Eucalyptus essential oil, were tested in Petri dishes on the germination and growth of Amaranthus retroflexus. The allelopathic effects of α-terpineol on A. retroflexus were examined with a pot experiment by measuring germination rate and biomass. The yield of essential oil was 0.04%. Ninety-two chemical components were detected from the essential oil. In Petri dish experiment, the germination inhibitory effect under 5 µL and 7.5 µL of α-terpineol per dish reached 100%. In pot experiment, germination, aboveground fresh and dry weights of A. retroflexus were significantly affected by α-terpineol. At the concentration of 1.6 µL·cm-3, the inhibition effect was strongest, with the allelopathic response index being -0.51 for germination, -0.62 for aboveground fresh weight, and -0.44 for aboveground dry weight, and the inhibition ratio being 51%, 62% and 44%, respectively.

[Study on literature of artichoke and properties of traditional Chinese medicine].

In this article, the foreign and domestic literature on alien plant medicine Cynara scolymus was reviewed to explore its properties and functions in traditional Chinese medicine theory, and provide theoretical basis for clinical application and reasonable compatibility. Based on the literature databases of PubMed, Web of Science, Embase, the Cochrane Library, CNKI, VIP and Sinomed, the articles with high reliability related to C. scolymus were screened out and the obtained articles were systematically classified according to clinical application, chemical compositions, pharmacological action, toxic and side effects, etc. In the analysis with traditional Chinese medicine theory, it is concluded that: C. scolymus tastes bitter and slightly cold, attributing to spleen, stomach, liver and gall meridians. It has the functions of eliminating accumulation and guiding stagnation, regulating Qi-flowing for harmonizing stomach, clearing away dampness and heat, resolving turbidity and lowering blood lipids. It can be used for the treatment of dyspepsia, diet reduction, vomiting, nausea, abdominal distention, hypochondriac pain, jaundice, hyperlipidemia, etc. Through the analysis and research of the relevant literature on C. scolymus, the properties and functions of the drug were clarified, which could provide a theoretical basis for further animal experiments and clinical research. The research model of "traditional Chinese medicine theory" for alien plant medicines can provide reference for the introduction and research of botanical drugs around the world, which can greatly enrich Chinese medicine resources and is of great significance for promoting the sustainable development of traditional Chinese medicine.

Impact of early diagnosis of carotid artery stenosis by carotid ultrasound: A protocol of systematic review and meta-analysis.

BACKGROUND: The purpose of this study is to explore the impact of carotid ultrasound (CU) for early diagnosis of carotid artery stenosis (CAS). METHODS: Literatures will be sought from the following electronic databases: MEDLINE, EMBASE, Cochrane Library, PSYCINFO, Web of Science, Allied and Complementary Medicine Database, and China National Knowledge Infrastructure. The search will cover from the start of indexing to the present without any limitations of language and publication status. All study quality will be assessed by Quality Assessment of Diagnostic Accuracy Studies tool, and data will be analyzed by RevMan V.5.3 software and Stata V.12.0 software. RESULTS: This study will investigate the impact of CU for early diagnosis of CAS through sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, and diagnostic odds ratio. CONCLUSION: The findings of this study may provide helpful evidence for the impact of CU for early diagnosis of CAS. SYSTEMATIC REVIEW REGISTRATION: PROSPERO CRD42019153904.

Two novel polysaccharides from rhizomes of Cibotium barometz promote bone formation via activating the BMP2/SMAD1 signaling pathway in MC3T3-E1 cells.

Cibotium barometz, an important traditional Chinese medicine, is used in strengthening bones and tendons. We found that C. barometz crude polysaccharides (CB70) could alleviate bone loss and markedly improve the biomechanical properties of OVX rats. Thus, to clarify biological active ingredient(s) of CB70, two homogeneous polysaccharides (CBP70-1-1 and CBP70-1-2) were purified from CB70. A combination of monosaccharide composition, FT-IR, GC-MS and NMR analysis indicated that CBP70-1-1 was composed of →6)-D-Galp-(1→, D-Glcp-(1→, →3,6)-D-Manp-(1→, →4)-D-Glcp-(1→ and →6)-D-Glcp-(1→ with relative molecular weights of 12,724 Da, and CBP70-1-2 was composed of →4)-D-Glcp-(1→, D-Glcp-(1→, →3,6)-D-Manp-(1→, →6)-D-Galp-(1→, →4,6)-D-Glcp-(1→ and →3)-L-Araf-(1→ with relative molecular weights of 3611 Da. Morphological analyses revealed that CBP70-1-1 and CBP70-1-2 appeared as a sheet that were irregular in size and shape, while the surface of CBP70-1-1 was full of sharp protuberances and CBP70-1-2 was smooth. Furthermore, the effects of CBP70-1-1 and CBP70-1-2 on the proliferation, differentiation and mineralization of mouse pre-osteoblastic MC3T3-E1 cells were assessed via CCK-8 assay, alkaline phosphatase activity assay, and alizarin red-based assay, respectively. These results revealed that CBP70-1-1 and CBP70-1-2 significantly promoted the proliferation, differentiation and mineralization of MC3T3-E1 cells, even better than E2. More importantly, quantitative real-time PCR and Western blot analysis indicated that CBP70-1-2 pronouncedly promoted the expression of osteogenic-related marker genes (Runx2, Osx, Ocn and Opn) and proteins (BMP2, RUNX2, OSX and p-SMAD1), which implies that the osteogenic activity of CBP70-1-2 is accomplished mainly by activating the BMP2/SMAD1 signaling pathway. These findings suggest CBP70-1-2 as a potential natural anti-osteoporotic agent for pharmacotherapy.

Structural identification and osteogenic activity of a novel heteropolysaccharide obtained from female flowers of Humulus lupulus.

Humulus lupulus is distributed mainly in Asia, Europe, and North America, and the flowers and infructescences are used worldwide in beer brewing as an essential flavoring ingredient. In China, the main producing areas are the Xinjiang Uygur Autonomous Region and Gansu provinces. The annual production of H. lupulus in China accounts for about 13% of worldwide production, ranking third after the United States and Germany. The female flower of H. lupulus also is used in traditional Chinese medicine, but the bioactive components responsible for its benefits remain unclear. In this study, we isolated a homogeneous polysaccharide (HLP50-2) from female flowers of H. lupulus via DEAE-52 cellulose and Sephadex G-75 gel filtration chromatography. The molecular weight was 73.25 kDa. Chemical methods, gas chromatography-mass spectrometry, and nuclear magnetic resonance analyses showed that HLP50-2 consisted of →4)-ß-d-GlcAp-(1→, →4)-α-l-Rhap-(1→, →5)-α-l-Araf-(1→, α-l-Araf-(1→, →3,6)-ß-d-Galp-(1→, α-d-Galp-(1→, →3,4,6)-ß-d-Galp-(1→, ß-d-Xylp-(1→, →6)-α-d-Galp-(1→, and →4)-ß-d-Galp-(1→. Advanced structural analysis indicated that HLP50-2 has a sheet-like structure with uneven distribution and the surface was smooth. In addition, we used MC3T3-E1 cells to evaluate the osteogenic activity of HLP50-2. The results show that HLP50-2 significantly promoted the proliferation, differentiation, and mineralization of MC3T3-E1 cells at concentrations of 12.5 µg mL-1 and 25 µg mL-1. In particular, at 25 µg mL-1, the promoting mineralization effect of HLP50-2 was better than that of the positive control, 17ß-estradiol, and the alkaline phosphatase activity after 10 d of treatment was comparable to that of the positive control. The extraction, isolation, purification, structural identification, and osteogenic activity of H. lupulus polysaccharides suggest that homogeneous polysaccharide HLP50-2 has pronounced anti-osteoporosis activity. Therefore, it has broad application prospects in pharmaceutical industries and health-care products.