ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
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.