Hawthorn leaf flavonoids alleviate the deterioration of atherosclerosis by inhibiting SCAP-SREBP2-LDLR pathway through sPLA2-â ¡A signaling in macrophages in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Hawthorn leaves are a combination of the dried leaves of the Rosaceae plants, i.e., Crataegus pinnatifida Bge. or Crataegus pinnatifida Bge. var. major N. E. Br., is primarily cultivated in East Asia, North America, and Europe. hawthorn leaf flavonoids (HLF) are the main part of extraction. The HLF have demonstrated potential in preventing hypertension, inflammation, hyperlipidemia, and atherosclerosis. However, the potential pharmacological mechanism behind its anti-atherosclerotic effect has yet to be explored. AIM OF THE STUDY: The in vivo and in vitro effects of HLF on lipid-mediated foam cell formation were investigated, with a specific focus on the levels of secreted phospholipase A2 type IIA (sPLA2-II A) in macrophage cells. MATERIALS AND METHODS: The primary constituents of HLF were analyzed using ultra-high performance liquid chromatography and liquid chromatography-tandem mass spectrometry. In vivo, HLF, at concentrations of 5 mg/kg, 20 mg/kg, and 40 mg/kg, were administered to apolipoprotein E knockout mice (ApoE-/-) fed by high-fat diet (HFD) for 16 weeks. Aorta and serum samples were collected to identify lesion areas and lipids through mass spectrometry analysis to dissect the pathological process. RAW264.7 cells were incubated with oxidized low-density lipoprotein (ox-LDL) alone, or ox-LDL combined with different doses of HLF (100, 50, and 25 µg/ml), or ox-LDL plus 24-h sPLA2-IIA inhibitors, for cell biology analysis. Lipids and inflammatory cytokines were detected using biochemical analyzers and ELISA, while plaque size and collagen content of plaque were assessed by HE and the Masson staining of the aorta. The lipid deposition in macrophages was observed by Oil Red O staining. The expression of sPLA2-IIA and SCAP-SREBP2-LDLR was determined by RT-qPCR and Western blot analysis. RESULTS: The chemical profile of HLF was studied using UPLC-Q-TOF-MS/MS, allowing the tentative identification of 20 compounds, comprising 1 phenolic acid, 9 flavonols and 10 flavones, including isovitexin, vitexin-4″-O-glucoside, quercetin-3-O-robibioside, rutin, vitexin-2″-O-rhamnoside, quercetin, etc. HLF decreased total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and non-high-density lipoprotein cholesterol (non-HDL-C) levels in ApoE-/- mice (P < 0.05), reduced ox-LDL uptake, inhibited level of inflammatory factors, such as IL-6, IL-8, TNF-α, and IL-1ꞵ (P < 0.001), and alleviated aortic plaques with a thicker fibrous cap. HLF effectively attenuated foam cell formation in ox-LDL-treated RAW264.7 macrophages, and reduced levels of intracellular TC, free cholesterol (FC), cholesteryl ester (CE), IL-6, TNF-α, and IL-1ß (P < 0.001). In both in vivo and in vitro experiments, HLF significantly downregulated the expression of sPLA2-IIA, SCAP, SREBP2, LDLR, HMGCR, and LOX-1 (P < 0.05). Furthermore, sPLA2-IIA inhibitor effectively mitigated inflammatory release in RAW264.7 macrophages and regulated SCAP-SREBP2-LDLR signaling pathway by inhibiting sPLA2-IIA secretion (P < 0.05). CONCLUSION: HLF exerted a protective effect against atherosclerosis through inhibiting sPLA2-IIA to diminish SCAP-SREBP2-LDLR signaling pathway, to reduce LDL uptake caused foam cell formation, and to slow down the progression of atherosclerosis in mice.

Uncovering the Mechanism of Chinese Hawthorn Leaf on Myocardial Ischemia Based on Network Pharmacology, Molecular Docking Verification, and In Vitro Studies.

Hawthorn leaf has shown therapeutic effects in the patients with myocardial ischemia. Our study combines network pharmacology, molecular docking techniques, and in vitro experiment with the aim of revealing the mechanism of hawthorn leaves in the treatment of myocardial ischemia. The active ingredients and corresponding targets of hawthorn leaf through Traditional Chinese Medicine System Pharmacology and Swiss Target Prediction databases. Targets related to myocardial ischemia were retrieved by Gene Card, Online Mendelian Inheritance in Man, Disgenet, and Therapeutic Targets Database databases. Cytoscape software was used to construct an ingredient-target-organ network and enrichment analysis of common targets was analyzed. Molecular docking verification of the core compound and target interactions was performed using MOE software. In vitro cell experiment was performed to verify the findings from bioinformatics analysis. Six active components and 107 potential therapeutic targets were screened. The protein-protein interaction network analysis indicated that 10 targets, including AKT1 and EGFR, were hub genes. Quercetin, kaempferol and isorhamnetin were taken as core active components. Through pathway enrichment analysis, nearly 455 Gene Ontology entries and 77 Kyoto Encyclopedia of Genes and Genomes pathways were obtained, mainly including PI3K/Akt, estrogen and other signaling pathways. Molecular docking prediction showed that three main active ingredients were firmly combined with the core targets. Cellular experiments showed that quercetin alleviated oxidative damage in cells and regulated the expression of PI3K, P-AKT/AKT and Bax/Bcl-2 proteins. This study identified the potential targets of Hawthorn leaf against myocardial ischemia using network pharmacology and in vitro verification, which provided a new understanding of the pharmacological mechanisms of Hawthorn leaf in treatment of myocardial ischemia.

An Interpretable Screening Approach Derived Through XGBoost Regression for the Discovery of Hypolipidemic Contributors in Chinese Hawthorn Leaf and its Counterfeit Malus Doumeri Leaf.

The active ingredient group is a prominent feature reflecting the inherent characteristics of plant-based functional foods. Chinese hawthorn leaf (CHL), a tea substitute possessing intrinsic nutritional properties in anti-hyperlipidemia, was first found to be adulterated with Malus doumeri leaf (MDL) owing to similar commercial labels. In this context, the above-mentioned two contrasting species were explored through phytochemical profiling and activity assessment. The amelioration effect of CHL on free fatty acids-elicited lipid deposition in HepG2 cells was significantly better than that of MDL. Molecular networking-based metabolic profiles identified 68 and 67 components in CHL and MDL, with 33 shared components. Extreme gradient boosting (XGBoost) algorithm with outstanding performance was selected to screen candidate components contributing to hypolipidemic activity, and the output was later interpreted by Shapley additive explanations (SHAP) method. Twelve and eight components were separately screened as hyperlipidemic inhibitors in CHL and MDL, while only four constituents were shared. The bioactivity evaluation of selected ingredients and combinations further confirmed their anti-hyperlipidemia capacity. These findings emphasized the feasibility of filtering bioactivity-related compounds using interpretable machine learning approaches and illustrated that related species may contain different hypolipidemic contributors, even if shared constituents existed.

Neuroprotective effects of hawthorn leaf flavonoids in Aß25-35 -induced Alzheimer's disease model.

Alzheimer's disease (AD) is a progressive neurodegenerative disease characterized by ß-amyloid (Aß) plaques, neurofibrillary tangles, neuronal cell loss, and oxidative stress. Further deposition of Aß in the brain induces oxidative stress, neuroinflammation, and memory dysfunction. Hawthorn (Crataegus pinnatifida Bge.) leaf, a known traditional Chinese medicine, is commonly used for the treatment of hyperlipidemia, heart palpitations, forgetfulness, and tinnitus, and its main bioactive components are Hawthorn Leaf Flavonoids (HLF). In this study, we investigated the neuroprotective effects of the HLF on the Aß25-35 (bilateral hippocampus injection) rat model of AD. The results showed that the oral administration of HLF at a dose of 50, 100, and 200 mg/kg for 30 days significantly ameliorated neuronal cell damage and memory deficits, and markedly increased the enzyme activities of superoxide dismutase and catalase, and the content of glutathione whereas it decreased the malondialdehyde content in the Aß25-35 rat model of AD as well as suppressed the activation of astrocytes. In addition, HLF up-regulated Nrf-2, NQO-1, and HO-1 protein expressions. Also, it reduced neuroinflammation by inhibiting activation of astrocytes. In summary, these results indicated that HLF decreased the oxidative stress via activating Nrf-2/antioxidant response element signaling pathways, and may suggest as a potential candidate for AD therapeutic agent.

High-throughput LC-MS method for the rapid characterisation and comparative analysis of multiple ingredients of four hawthorn leaf extracts.

INTRODUCTION: The comprehensive component characterisation of Chinese herbal medicine is the premise of effectively driving the discovery of pharmacodynamic substances or new drugs in recent years. OBJECTIVE: To use the high-throughput liquid chromatography-mass spectrometry (LC-MS) approach to systematically characterise phytochemical compounds from four hawthorn leaf extracts, along with evaluating their classification. METHODS: In the present study, the compounds from 50% ethanol extract, macro porous resin extract, ethyl acetate extract and standard decoction of hawthorn leaves were completely analysed by ultrahigh-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS). RESULTS: Eight-nine compounds were putatively identified by comparison with secondary MS data and available references. Of these compounds identified, 56 compounds were found for the first time in hawthorn leaves, which was somewhat inconsistent with the findings of other studies. It could be inferred that falconoid, organic acids and nitrogenous compounds were the most abundant in 50% ethanol extract and standard decoction extract, which were considered as better choices for extracting hawthorn leaves. CONCLUSIONS: This work developed a simple, accurate and rapid method for the compound identification of hawthorn leaves, which laid the basis for further discovering pharmacodynamic material basis or new drugs from hawthorn leaves.

Comparative analysis of chemical constituents in hawthorn leaves from different sources / 药学学报

We identified and analyzed the components and chemical constituents of hawthorn leaves of Crataegus pinnatifida Bge. (wild) and C. pinnatifida Bge. var major N. E. Br (cultivated) by using ultra high-performance liquid chromatography and quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MSE) combined with UNIFI data analysis platform and multivariate statistics. Fifty-eight chemical compounds were identified, including organic acids, flavonoids, triterpenoic acids, monoterpenes and sesquiterpenoids; among them, terpenoid content was the most abundant. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to identify the differential components of hawthorn leaves from two sources. The results show that there are differences in the chemical compositions of the two sources, including 24 flavonoids and terpenoids (including monoterpenoids, sesquiterpenoids and triterpenoid acids). The types of flavonoids (such as rutin, vitexin-2''-O-rhamnoside, isovitexin-2''-O-rhamnoside, hyperoside, quercetin-3-O-β-D-glucoside) and terpenoids (crataegolic acid, corosolic acid and ursolic acid) in C. pinnatifida were more varied than those found in C. pinnatifida Bge. var major N. E. Br, and their contents were relatively higher. This study provides a comprehensively analysis of the different chemical components of hawthorn leaves from two sources listed in the Chinese Pharmacopoeia, and provides a basis for the selection of raw materials and the potential development and utilization of hawthorn leaves.

Ecofriendly microwave-assisted reaction and extraction of bioactive compounds from hawthorn leaf.

INTRODUCTION: The main active components in hawthorn leaves possess various biological activities such as anti-inflammatory, antioxidant, and hypolipidemic effects. Therefore, it is necessary to develop an effective and reliable extraction method to extract these active compounds from hawthorn leaves. OBJECTIVE: To establish a simple, rapid, and sensitive method for extraction and determination of polyphenolic compounds from hawthorn leaves. METHODS: In this study, a microwave-assisted reaction and extraction (MARE) combined with ultra-high-performance liquid chromatography with ultraviolet detector method was established to extract and determine the polyphenolic compounds in hawthorn leaves. The solid reagent aqueous solutions were applied as extraction solvents, preventing the use of organic solvents. The target analytes were identified by quadrupole time-of-flight tandem mass spectrometry. Several experimental parameters that can significantly affect the extraction efficiency were evaluated and optimised. RESULTS: The optimal conditions were as follows: 0.1 g of sodium carbonate was used as solid reagent, the amount of sodium borate was set at 0.01 g, extraction time was 10 min, extraction temperature was set at 50°C, pH value was adjusted to 7. The validation experiments demonstrated that the method had high sensitivity with the limits of detection in the range 26.5-37.7 ng/mL. The average recoveries ranged from 80.22% to 93.27%. CONCLUSION: In this work, the proposed MARE method was successfully applied to extract and determine polyphenolic compounds in hawthorn leaf samples. Compared with other reported methods, the present method was faster, greener, and more sensitive.

Effect of Hawthorn Leaf Flavonoids in Dehydroepiandrosterone-Induced Polycystic Ovary Syndrome in Rats.

OBJECTIVE: To evaluate the potential beneficial effects of hawthorn leaf flavonoids (HLF) against polycystic ovary syndrome (PCOS) in a rat model of disease and elucidate the underlying molecular mechanism. METHODS: The PCOS model was established by subcutaneous injection of dehydroepiandrosterone (DHEA, 60 mg/kg/day) for 21 consecutive days. HLF (200 mg/kg/day) were orally administered simultaneously or after the injection. The body weight was regularly monitored and recorded. The ovaries were weighed and histologically examined via hematoxylin and eosin staining. The number of follicular cysts was counted under a light microscope. The serous hormones were measured using enzyme-linked immunosorbent assay kits. Insulin resistance (IR) was calculated as HOMA-IR = fasting insulin (µU/L) × fasting glucose (mM)/22.5. The estrous cycle was determined by vaginal smear. The relative expression of tumor necrosis factor-α and interleukin-6 was measured by real-time polymerase chain reaction. The superoxide dismutase activity and malondialdehyde content was determined using commercially available kits. RESULTS: DHEA induced a significant increase of body weight, ovary weight, number of follicular cysts, serous hormones, IR, inflammatory cytokines, and oxidative stress, and it also impaired the estrous cycle. Oral administration of HLF greatly alleviated these complications. Little toxicity of HLF was observed in our rat model. CONCLUSION: HLF manifest protective effects against PCOS progression in the animal model, which may hold great promise for future clinical applications.

[Calcium mobilizing effect of hawthorn leaf procyanidins in vascular endothelial cells].

The hawthorn leaves have the effect of activating blood, removing blood stasis, regulating qi through the veins, dissolving turbidity and lowering lipid. Procyanidinis is one of its main active components and plays an important role in regulating vasoactivity. Previous studies showed that the regulating effect of procyanidins was related to its regulation on nitric oxide secretion from vascular endothelial cells, and this effect was dependent on the extracellular calcium concentration, suggesting that the changes in intracellular calcium ion concentration in endothelial cells may play a key role in this process. However, the research on this issue is still insufficient so far. This study is aimed to observe the effect of hawthorn leaf oligomeric procyanidins (HLP) on calcium mobilization of vascular endothelial cells, and investigate the underlying mechanism. Human umbilical vein endothelial cells (HUVEC) were cultured in vitro and labeled with Fura-2. HUVEC were treated with HLP at concentrations of 6.25, 12.5, 25 and 50 mg·L⁻¹, and the intracellular calcium concentrations were measured with a living cell microscope for 30 min. HLP increased the intracellular calcium concentration of HUVEC in a concentration dependent manner; and the intracellular calcium concentrations in 25 and 50 mg·L⁻¹ HLP groups were significantly higher than that in the normal group. With the use of calcium-free incubation buffer, addition of calcium chelating agent EGTA in incubation buffer, or use of inhibitors for sodium calcium exchanger, the effect of HLP was significantly inhibited. On the other hand, the effect of HLP could also be weakened by inhibiting the calcium release from the intracellular storage. In conclusion, these results suggest that HLP can elicit calcium mobilization in vascular endothelial cells, which may be one of the mechanisms for its vascular modulatory activity; and this calcium mobilizing effect may be achieved through promoting both extracellular calcium influx and intracellular calcium release, additionally the former may be related to activating the reverse transport of Na⁺-Ca²âº exchangers on the cell membrane.

Calcium mobilizing effect of hawthorn leaf procyanidins in vascular endothelial cells / 中国中药杂志

The hawthorn leaves have the effect of activating blood, removing blood stasis, regulating qi through the veins, dissolving turbidity and lowering lipid. Procyanidinis is one of its main active components and plays an important role in regulating vasoactivity. Previous studies showed that the regulating effect of procyanidins was related to its regulation on nitric oxide secretion from vascular endothelial cells, and this effect was dependent on the extracellular calcium concentration, suggesting that the changes in intracellular calcium ion concentration in endothelial cells may play a key role in this process. However, the research on this issue is still insufficient so far. This study is aimed to observe the effect of hawthorn leaf oligomeric procyanidins (HLP) on calcium mobilization of vascular endothelial cells, and investigate the underlying mechanism. Human umbilical vein endothelial cells (HUVEC) were cultured and labeled with Fura-2. HUVEC were treated with HLP at concentrations of 6.25, 12.5, 25 and 50 mg·L⁻¹, and the intracellular calcium concentrations were measured with a living cell microscope for 30 min. HLP increased the intracellular calcium concentration of HUVEC in a concentration dependent manner; and the intracellular calcium concentrations in 25 and 50 mg·L⁻¹ HLP groups were significantly higher than that in the normal group. With the use of calcium-free incubation buffer, addition of calcium chelating agent EGTA in incubation buffer, or use of inhibitors for sodium calcium exchanger, the effect of HLP was significantly inhibited. On the other hand, the effect of HLP could also be weakened by inhibiting the calcium release from the intracellular storage. In conclusion, these results suggest that HLP can elicit calcium mobilization in vascular endothelial cells, which may be one of the mechanisms for its vascular modulatory activity; and this calcium mobilizing effect may be achieved through promoting both extracellular calcium influx and intracellular calcium release, additionally the former may be related to activating the reverse transport of Na⁺-Ca²⁺ exchangers on the cell membrane.

Effects of hawthorn leaf polymeric procyanidins on calcium mobilization in human umbilical vein endothelial cells / 中国病理生理杂志

AIM:To observe the effects of hawthorn leaf polymeric procyanidins ( PPC) on calcium mobiliza-tion of vascular endothelial cells , and to study the underlying mechanism .METHODS: Free calcium in cultured human umbilical vein endothelial cells (HUVECs) was labeled with Fura-2.HUVECs were treated with ATP, a positive control drug, and PPC at concentrations of 12.5, 25 and 50 mg/L..The intracellular calcium concentrations were measured with a living cell microscope for 30 min.RESULTS:PPC concentration-dependently increased the intracellular calcium concen-tration of HUVECs .The intracellular calcium concentrations in 25 and 50 mg/L PPC groups were significantly higher than that in normal group (P<0.01).The dynamic manner of calcium concentration elevations elicited by PPC was a slow in -crease which happened after a latency time of several minutes , lasted for several minutes , and reached a plateau finally . This manner was quite different from that elicited by ATP , a typical SOC operator , hinting different mechanisms between them .Inhibiting the intracellular calcium release did not influence the effects of PPC;however , deleting extracellular calci-um, inhibiting the reverse mode of Na +-Ca2+exchange, or deleting extracellular sodium , restrained or even abolished the effects of PPC.CONCLUSION:PPC elicits calcium mobilization in vascular endothelial cells , which may be one of the mechanisms of the vascular modulatory activity of hawthorn procyanidins .This effect may be achieved through inducing the influx of sodium and then activating the reverse mode of Na +-Ca2+exchange.

Effects of Hawthorn Leaf Procyanidins on Over Expressions of ICAM-1 and E-selectin in Human Umbilical Vein Endothelial Cells Induced by TNF-α / 中国中医药信息杂志

Objective To observe the effects of hawthorn leaf procyanidins (HLP) on over expressions of ICAM-1 and E-selectin in human umbilical vein endothelial cells (HUVEC) induced by TNF-α,and clarify the mechanism of HLP's anti-inflammation effect. Methods HUVEC were cultured in vitro. MTT assay was used to detect cell viabilities. The expressions of ICAM-1 and E-selectin in HUVEC were detected by flowcytometry. Results Up to 200 mg/L, HLP showed no significant decrease in cell viabilities; the expression levels of ICAM-1 and E-selectin in the model group significantly increased, compared with that in the normal group; 10, 20, 30, 40, 50 mg/L HLP inhibited the expression elevations of ICAM-1 and E-selectin in concentration-dependent manner; and there were statistical significances in 40, 50 mg/L HLP groups, compared with the model group. Conclusion HLP can inhibit the over expressions of ICAM-1 and E-selectin of vascular endothelial cells induced by TNF-α, which possibly underlies HLP's anti-inflammation effect.

Effects of total flavone of hawthorn leaf on expression of p38MAPK signaling pathway and inflam-mation factors in rats brain with chronic cerebral ischemia / 中华行为医学与脑科学杂志

Objective To investigate the therapeutic effect and mechanism of total flavone of haw-thorn leaf ( TFHL) on p38MAPK signaling pathway and inflammation factors in rats brain with chronic cere-bral ischemia.Methods SPF class healthy male SD rats were randomly divided into sham group, model group,TFHL group and Ginkgo leaf group( 12 rats in each group) .Permanent bilateral carotid artery ligation was used to prepare chronic cerebral ischemia model.Morris water maze method was used to evaluate learn-ing and memory abilities of rats.Immunohistochemistry and Western blot methods were used to measure the expression of caspase-3 and p38MAPK proteins.ELISA method was used to measure the amounts of TNF-αand IL-1βin hippocampal tissue.Results Compared with the model group,TFHL treatment (36 d) can im-prove learning and memory capabilities of vascular dementia rats,shorten the escape latency ( TFHL group(10.01±2.85) s vs Model group (19.54±6.12) s, P<0.05) and the course of searching platform(TFHL group(2.6044±0.3219)m vs model group(3.3502±0.6231)m, P<0.05),increase the numbers of crossing the platform (TFHL group(5.17±2.12) times vs Model group (3.96±1.34) time,s P<0.05) and the platform quadrant swimming distance percentage (TFHL group(48.22±7.39)%vs model group (33.42±5.32) %, P<0.01).The number of caspase-3 positive cells in the hippocampus significantly reduced (TFHL group(1.677 ±0.164) vs Model group (2.387±0.171), P<0.05),the expression level of P38MAPK protein (TFHL group (0.0161±0.0003) vs Model group (0.0254±0.0018), P<0.05),TNF-α(TFHL group(19.61±3.61) ng/10 mg vs Model group (27.82±6.57) ng/10 mg, P<0.01)and IL-1β(TFHL group(24.41±2.56) ng/10 mg vs Model group (29.43±5.26) ng/10 mg, P<0.05) were significantly decreased.Conclusion TFHL plays a protective role in nerve function of the chronic cerebral ischemia rats.The mechanism of its antia-poptosis might be associated with the activation of P 38MAPK signaling pathway,inflammation and the apoptosis of neurons in the brain.

Hawthorn leaf flavonoids alleviate nonalcoholic fatty liver disease by enhancing the adiponectin/AMPK pathway.

Hawthorn (Crataeguspinnatifida) belongs to the genus Rosaceae family of plants. The hawthorn leaf, Crataeguspinnatifida Bunge, is used for both condiment and medicinal purposes to prevent and treat metabolic dysfunctions, such as hyperlipidemia, hypertension, and cardiovascular disease in traditional Chinese medicine. However, its effects on nonalcoholic fatty liver disease (NAFLD) remain obscure. The purpose of the present study was to investigate the protective effect of hawthorn leaf flavonoids (HLF), the dominant bioactive extracts of hawthorn leaves, on high fat diet (HFD)-induced hepatic steatosis and to elucidate its underlying mechanisms. HLF supplementation significantly lowered body weight, liver weight, liver/body weight ratio, improved serum parameters and liver dysfunction and markedly decreased hepatic lipid accumulation in HFD-fed rats. In addition, HLF intervention dramatically increased circulating adiponectin levels and up-regulated the expression of adiponectin receptors, particularly adiponectin receptor 2 (AdipoR2) in the liver. Moreover, adenosine monophosphate (AMP)-activated protein kinase (AMPK) was also activated, as well as AMPK-mediated alteration of sterol regulatory element binding protein-1c (SREBP-1c), peroxisome proliferator-activated receptor α (PPARα) and their downstream targets. Taken together, our data suggest that HLF ameliorates hepatic steatosis by enhancing the adiponectin/AMPK pathway in the liver of HFD-induced NAFLD rats.

Tissue distribution comparison between healthy and fatty liver rats after oral administration of hawthorn leaf extract.

Hawthorn leaves, a well-known traditional Chinese medicine, have been widely used for treating cardiovascular and fatty liver diseases. The present study aimed to investigate the therapeutic basis treating fatty liver disease by comparing the tissue distribution of six compounds of hawthorn leaf extract (HLE) in fatty liver rats and healthy rats after oral administration at first day, half month and one month, separately. Therefore, a sensitive and specific HPLC method with internal standard was developed and validated to determine chlorogenic acid, vitexin-4''-O-glucoside, vitexin-2''-O-rhamnoside, vitexin, rutin and hyperoside in the tissues including heart, liver, spleen, kidney, stomach and intestine. The results indicated that the six compounds in HLE presented some bioactivity in treating rat fatty liver as the concentrations of the six compounds varied significantly in inter- and intragroup comparisons (healthy and/or fatty liver group).

Nutritional and functional potential of Beta vulgaris cicla and rubra.

Swiss chard (Beta vulgaris cicla, BVc) and beetroot (Beta vulgaris rubra, BVr) are vegetables of the Chenopodiaceae family, widely consumed in traditional western cooking. These vegetables represent a highly renewable and cheap source of nutrients. They can be cultivated in soils with scarce organic material and little light and water. BVc and BVr have a long history of use in folk medicine. Modern pharmacology shows that BVc extracts possess antihypertensive and hypoglycaemic activity as well as excellent antioxidant activity. BVc contains apigenin flavonoids, namely vitexin, vitexin-2-O-rhamnoside and vitexin-2-O-xyloside, which show antiproliferative activity on cancer cell lines. BVr contains secondary metabolites, called betalains, which are used as natural dyes in food industry and show anticancer activity. In this light, BVc and BVr can be considered functional foods. Moreover, the promising results of their phytochemicals in health protection suggest the opportunity to take advantage of the large availability of this crop for purification of chemopreventive molecules to be used in functional foods and nutraceutical products.

Effects of hawthorn leaf flavonoids on reducing blood lipids and preventing fatty liver in the quails / 复旦学报（医学版）

Objective To investigate the effect of hawthorn leaf flavonoids (HLFs) with the different concentrations in prevention of high blood lipids and fatty liver in quails. Methods Sixty quails were divided into six groups:normal (N),high lipids food model (M),positive control (ZBT),HLF1,HLF2 and HLF3 groups.The animals in the N group were fed with normal food,while the M group was feed with high lipids food to establish the animal model of fatty liver.Besides high-fat diet,HLF1,HLF2 and HLF3 groups were also fed with 1 mL HLFs in the doses of 20 mg/kg,40 mg/kg and 80 mg/kg respectively.The ZBT group was fed with 1 g/kg of ZhiBiTuo and used as the positive control.After the experiments of six months,the serum and liver of the quails were taken and used to evaluate the contents of total cholesterol (TC),triglyceride (TG),high density lipid cholesterol(HDL-C),low density lipid cholesterol (LDL-C),superoxide dismutase (SOD),malondialdehyde (MDA),glutathione (GSH),alanine amino transferase (ALT) and glutamic oxaloacetic transaminase (AST).Meanwhile,the microscopic and electronic microscopic sections were used to observe the pathological changes of the livers of the quails. Results Compared with the M group,HLFs significantly lowered the content of TC,TG,LDL-C,MDA,AST and ALT in the serum and livers of the quails in other groups (P<0.001).However,the levels of HDL-C,SOD and GSH of the other groups were increased markedly (P<0.001).Moreover,the ratios of increase/decrease in the three HLF groups were depended on the concentrations of HLFs.Also,better liver structure and less lipid accumulation were also observed in the HLF groups when compared to the M group. Conclusions These results showed HLFs can lower blood lipid and cholesterol accumulation,improve the antioxidant capacity and protect the physiological function of liver.So,HLFs were effective in preventing and treating high blood lipids and fatty liver induced by high-fat diet.

Effect of total flavonoids of Chinese hawthorn leaf on expression of NF-?B and its inhibitor in rat liver with non-alcoholic steato-hepatitis / 中华中医药杂志

Objective:To study the effect of NF-?B and its inhibitor in rats liver during the development of non-alcoholic steato-hepatitis and observe the effect of total flavonoids of Chinese hawthorn leaf on expressions of NF-?B and its inhibitor,and explore the prevention and treatment mechanism of non-alcoholic steato-hepatitis. Methods:Non-alcoholic steato-hepatitis SD rats model were established by administering a high-fat diet for 12 weeks and were treated with total flavonoids of Chinese hawthorn leaf at dosages of 250mg/kg body weight /d,125 mg/kg body weight /d and with Essentiale at a dosage of 195.4mg /kg body weight /d. Pathological changes of liver tissues were observed with HE,the serum MDA,SOD and TNF-? were measured,the protein and mRNA expressions of NF-?B and I?B in rats liver were detected with RT-PCR and immunohistochemical method. Results:There were severe steatosis inflammatory cell infiltration and necrotic foci in non-alcoholic steato-hepatitis rat. Compared with the control group,contents of the serum MDA and TNF-? in non-alcoholic steato-hepatitis group obviously increased,the activity of SOD decreased,the mRNA and protein expressions of NF-?B P65 and I?B? in liver obviously strengthened. There was a positive relativity of the protein expression of NF-?B p65,I?B? with the contents of serum TNF-?,MDA,but a negative relativity with the activity of serum SOD. Conclusion:Total flavonoids of Chinese hawthorn leaf can obviously lower lipid peroxidation in rats,lower the harm of cytokine to hepatocytes,and regulate the expression of protein and mRNA of NF-?B and I?B. It maybe the important mechanism of total flavonoids of Chinese hawthorn leaf for preventing and treating the development of non-alcoholic steato-hepatitis.

Protective Effect of Total Flavones of Hawthorn Leaf on Renal Ischemia/Reperfusion Injury in Rats / 实用儿科临床杂志

Objective To study the protective effect of total flavones of hawthorn leaf (TFHL) on renal ischemia/reperfusion (I/R) in rats.Methods A model of renal I/R was made by clamping double renal pedical for 60 min and reperfusion for 24 h.TFHL (30 or 60 mg/kg) was injected before clamping renal ischemia. The level of serum urea nitrogen(BUN), serum creatinine(Scr), tumor necrosis factor-?(TNF-?),interleukin-1(IL-1) in serum were detected after reperfusion for 1 and 24 h,respectively,and the level of malonaldehyde (MDA) and erythrocuprein (SOD) in renal tissues were measured after reperfusion for 24 h.Results TFHL significantly decreased the level of BUN and Scr, decreased the concentration of TNF-? and IL-1 in serum. Pretreatment with TFHL also enhanced the activity of SOD reduced the concentration of MDA in renal after I/R.Conclusions TFHL has significantly protective effect on renal I/R and markedly improve the renal function. The mec-hanisms are associated with decreasing the lipid peroxidation reaction and reducing the level of TNF-? and IL-1.

Apparent partition coefficient of Hawthorn leaf flavonoids in phospholipids complex / 中成药

AIM: To explore the appearent partition coefficient of flavonoids from Hawthorn leaf in phospholids complex. METHODS: With the aid of the shake-flask method,The contents of rhamnosylvitexin,rutin and quercetin were determined by HPLC and apparent partition coefficient in the system of n-octyl alcohol and water of each ingredient in the physical mixture of Hawthorn leaf flavonoids and phospholipids,Hawthorn leaf flavonoids and its phospholipids complex were calculated respectively. RESULTS: Apparent partition coefficient of rhamnosylvitexin and rutin and quercetin in phospholipids complex were enhanced comparing with both in Hawthorn leaf flavonoids and the physical mixture of Hawthorn leaf flavonoids and phospholipids. CONCLUSION: Phospholipid complex may be an effective carrier to elevate the oral bioavailability of total flavonoids from effective fraction of Hawthorn leaf.