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@#AIM:To investigate the protective effects of Pollen Typhae extract on diabetic retinopathy(DR)in rats. <p>METHODS: Fifty SPF rats were randomly divided into five groups: control group with normal feeding, DR group was given the same amount of normal saline by gavage, experimental group A with Pollen Typhae extract 50mg/(kg·d), experimental group B with Pollen Typhae extract 100mg/(kg·d)and experimental group C with Pollen Typhae extract 200mg/(kg·d). Determination of fasting blood glucose in rats of each group. HE staining was used to observe the pathological changes of retina of rats in each group. The expression of IL-6 and TNF-α in the serum of rats were measured by ELISA. The VEGF, VEGFR2 and Ang-1 protein expression in retina tissue were observed by Western blot. The VEGF, VEGFR2 and Ang-1 mRNA expression in retinal tissue were observed by <i>q</i>RT-PCR. <p>RESULTS: Compared with the control group, the fasting blood glucose, the expression of IL-6 and TNF-α in serum and VEGF, VEGFR2 and Ang-1 protein and mRNA expression in retina tissue in DR group and each experimental group were significantly higher(<i>P</i><0.05). Compared with DR group, fasting blood glucose decreased in all experimental groups, and the fasting blood glucose of experimental groups B and C was significantly decreased than that of DR group(<i>P</i><0.05). In addition, the contents of IL-6 and TNF-α in serum, the protein and mRNA expression levels of VEGF, VEGFR2 and Ang-1 in retina tissue in experimental group B and group C were significantly lower than those in group DR(<i>P</i><0.05). The results of HE showed that the structure of retinal photoreceptor cell layer in DR group was obviously destroyed, the cell edema and gap widened, and the retinal histopathology of rat retina in group B and group C were improved in varying degrees. <p>CONCLUSION: Pollen Typhae extract can down-regulate the level of inflammation and the expression of VEGF, VEGFR2 and Ang-1 in DR rats, so as to improve the retinopathy.
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OBJECTIVE@#To test the hypothesis that the inhibition of endoplasmic reticulum (ER) stress-induced apoptosis in oxidized low-density lipoproteins (ox-LDL)-induced human aortic-vascular smooth muscle cells (HA-VSMCs) was associated with suppression of the protein kinase RNA-like ER kinase (PERK)-eukaryotic translation initiation factor 2α (eIF2α)-activating transcription factor 4 (ATF4)-CCAAT/enhancer binding protein homologous protein (CHOP) signaling pathway by Pollen Typhae total flavone (PTF).@*METHODS@#Primary HA-VSMCs were cultured and identified. The cultured HA-VSMCs were randomized into 5 groups, including a normal control group, an ox-LDL group (70 μg/mL high ox-LDL), an HPTF group (70 μg/mL high ox-LDL+500 μg/mL PTF), an MPTF group (70 μg/mL high ox-LDL+250 μg/mL PTF), and a LPTF group (70 μg/mL high ox-LDL+100 μg/mL PTF) in the first part; and a normal control group, an ox-LDL group (70 μg/mL high ox-LDL), an MPTF group (70 μg/mL high ox-LDL+250 μg/mL PTF), a shRNA group (transducted with PERK shRNA lentiviral particles), a scramble shRNA group (transducted with control shRNA lentiviral particles), an MPTF+ox-LDL+shRNA group (250 μg/mL PTF+70 μg/mL high ox-LDL+PERK shRNA lentiviral particles) and an ox-LDL+shRNA group (70 μg/mL high ox-LDL+PERK shRNA lentiviral particles) in the second part. The protein expression levels of ER-associated apoptosis proteins were detected by Western blot, and their mRNA expression levels were detected by quantitative real-time reverse transcription-polymerase chain reaction. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was applied to test cell viability, and the level of apoptosis was monitored by flow cytometry.@*RESULTS@#The MTT assay and flow cytometry showed that the ox-LDL group had a significant increase in apoptosis, which was attenuated in PTF treatment groups and shRNA groups. Moreover, the ox-LDL group had increased protein and mRNA levels of binding immunoglobulin protein and ER-associated apoptosis proteins, such as PERK, eIF2α, ATF4 and CHOP, which were attenuated in PTF treatment groups and shRNA groups.@*CONCLUSIONS@#The apoptosis induced by ox-LDL had a strong relation to ER stress. The protective effect of PTF on ER stressinduced apoptosis was associated with inhibition of the PERK-eIF2α-ATF4-CHOP pathway, which might be a potential therapeutic strategy for enhancing the stability of atherosclerotic plaques.
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Objective To study the effect of total flavonoids of Pollen Typhae on the autophagy of macro-phages and its effect on Akt/mTOR signaling pathway. Methods To observe the effect of serum containing drugs on the proliferation of SD rats. The serum containing 20%of the drug-containing serum and the macrophages were selected by the method of MMT. Interleukin(IL-10),interferon-γ(IFN-γ)and autophagy-related genes Akt and mTOR mRNA and protein were detected. Results There was no significant difference between the OD value of TYTF serum in the concentration range of 10% ~ 40% and so as the OD value of normal control(P > 0.05). The expression of autophagosomes in each dose group of TYTF was significantly increased under TEM. The autophagic expression was not typical in the normal control group. The expression of Beclin1 protein in TYTF group was signifi-cantly higher than that in normal control group(P 0.05). The expression of Beclin1 mRNA in TYTF group was significantly higher than that in normal control group (P < 0.05),while the expression of Akt and mTOR mRNA was significantly lower than that of normal control group (P < 0.05). The expression of IL-10 in TYTF group was significantly higher than that in normal control group (P < 0.05). The expression of IFN-γ was significantly lower than that of normal control group(P<0.05). Conclusions The total flavonoid serum has anti-atherosclerotic effect. The mechanism may be related to inhibition of the activation of Akt/mTOR signaling path-way,including macrophage autophagy activity induction ,plaque macrophage infiltration reduction and inflamma-tory response factors secretion inhibition ,inflammatory response activities inhibition. Thereby it can promote atherosclerosis plaque stability of the purpose of vulnerability.
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Objective To investigate the effects of Pollen Typhae total flavone (PTF) on INS-1 pancreaticβcell damage induced by palmitic acid ( PA) . Methods INS-1 pancreatic β cells were given long-term induction with PA to establish the impaired cell model, and then were intervened with PTF. Cell viability was determined by tetrazolium salt ( XTT) colorimetry. Results PA impaired the viabilities of INS-1 pancreatic β cells in concentration- and time-dependent manner, and PTF improved the impairment of INS-1 pancreatic β cells induced by PA in concentration -dependent manner. Moreover, PTF showed better improvement on the impairment when the INS-1 pancreatic β cells were impaired more seriously by PA. Conclusion PTF has effects on ameliorating the impairment of INS-1 pancreaticβcells induced by PA for long time.
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OBJECTIVE: To investigate the effect of compound pollen typhae extract on nephritis hematuria and renal function in rats, the pharmacodynamics evaluation laid a foundation for the development of hospital preparations compound pollen typhae granule. METHODS: The rat model of renal hematuria was induced by immunogen bovine serum albumin (BSA) gavages, carbon tetrachloride (CCl4) subcutaneous injection, and lipopolysaccharide (LPS) tail vein injection. Intragastric administration of high, medium and low-dose compound pollen typhae extract was performed for 6 weeks. Testing concentration of serum creatinine (SCr), urea nitrogen (BUN) and total protein (TP) in rat blood, and 24 h urine protein, urine creatinine quantitative, urine deformed erythrocyte number were detected in the fourth week and sixth week respectively. RESULTS: Compound pollen typhae extract reduced the urine deformed erythrocyte number, 24 h urinary proteins quantitative, cut down SCr and BUN, and increased TP and CCr, there are significant differences(P < 0.01 or P < 0.05). CONCLUSION: Compound pollen typhae extract is effective in the treatment of glomerulonephritis hematuria in rat model through improving renal function, with high dose group of best effect.
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Objective To investigate the effects of Pollen Typhae total flavone ( PTF) , an extract from Pollen Typhae which has the actions of activating blood and removing stasis, on inflammatory factors and insulin sensitivity in type 2 diabetic rats. Methods SD rats were used as the experimental animal. Type 2 diabetic rats induced by high fat diet plus low dose of streptozotocin were randomly divided into model group, PTF group (in the dosage of 200 mg·kg-1·d-1) , and rosiglitazone group (in the dosage of 4 mg·kg-1·d-1) . Additionally, normal control group was set up. After treatment for 4 weeks, plasma interleukin 6 ( IL-6) and tumor necrosis factor alpha ( TNF-α) levels were detected, the insulin tolerance test ( ITT) was performed, and the protein expression of suppressor of cytokine signaling-3 ( SOCS-3) in skeletal muscle was determined. Results After treatment for 4 weeks, the plasma levels of IL-6 and TNF-α, the homeostasis model of insulin resistance ( HOMA-IR) , and expression level of SOCS-3 in skeletal muscle in the model groups were significantly increased ( P﹤0.05) as compared with those in the normal control group, and insulin tolerance was also impaired in the model group ( P﹤0.05) . Compared with the model group, IL-6 level and HOMA-IR were markedly decreased in the PTF group ( P﹤0.05) , and the impaired insulin tolerance was obviously improved (P﹤0.05) . The level of SOCS-3 in the skeletal muscle of PTF group was also much lower than that of the model group and rosiglitazone group (P﹤0.05) . Conclusion PTF has effects on decreasing the levels of plasma IL-6 and SOCS-3 in the skeletal muscle and on improving insulin sensitivity in type 2 diabetic rats.
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Objective To establish a TLC identification method for pollen typhae in Xifeng-Tongnao capsule,and to set up quality control standard for the product.Methods According to the literatures,methods and improved methods were adopted to identify the pollen typhae in Xifeng-Tongnao capsule.Results The fifth method in this article was simple,with TLC spots being clear,the separation effect showing good results without the interference of negative control.Conclusion The fifth method has feasibility and a strong specificity.It can be used as qualitative identification method for pollen typhae in Xifeng-Tongnao capsule.
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bjective:To study and establish the fingerprint of Pollen Typhae.Methods:Hypersil BDS C18(5?m,4.6mm?250mm) chromatographic column mobile phase acetonitrile-0.1 % phosphoric acid solution(10:90)with fiow rate of 1.0 ml/min and UV detector at 254 nm.ResultsTaking Isorhamnetin-3-O-neohesperidoside as the reference peak,11 common peaks were selected as the fingerprint peaks of Pollen Typhae,Technology investigation indicated that the analytical method this study established has desirable precision,reproducibility,and stability.The similarity of Pollen Typhae fingerprints from different batches is better.Conclusion HPLC fingerprint analysis can be a method for quality control of medical material of Pollen Typhae.
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Alcohol-infusion. Conclusion: With the cell-wall-broken extraction process, a higher content of total flavones was obtained from Pollen Typhae .
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Objective To establish optimum processing method for Pollen Typhae Carbonisatus. Method Processing method was studied by orthogonal test and the total flavones were determined by HPLC. Conditions of HPLC used to determine the total flavones were: Waters Nove-Park C18 (150 mm?3.9 mm, 4 ?m), mobile phase: MeOH-THF-0.05%TFT (16∶24∶60), flow rate: 0.8 mL/min, column temperature: 30 ℃, detection wavelength: 360 nm. Results The optimum processing method was skir-baked for 8 min at 210 ℃. Conclusion The optimized processing method is available for the processing of Pollen Typhae Carbonisatus.
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Objective To establish the HPLC Fingerprint of Pollen Typhae and give a new method for quality control of Pollen Typhae. Methods RP-HPLC was used on a YWG-C_ 18 column (250 mm?4.6 mm, 5 ?m) with the gradient elution solvent system composed of acetonitrile-0.05% H_3PO_4 water solution, the detection wavelength was 254 nm, sample injection was 5 ?L. Results HPLC Fingerprint of Pollen Typhae was established. Taking typhaneoside as the reference peak, 12 common peaks were selected as the fingerprint peaks of Pollen Typhae, the similarity between the fingerprint of eight batches of Pollen Typhae samples and reference fingerprint was determined. Conclusion The established HPLC fingerprint has desirable precision, reproducibility, and stability, and can be applied to the quality control of Pollen Typhae.
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Objective Dissolution test was applied to study the dissolution rates of total flav ones from Pollen Typhae before and after wall breaking.Methods The content of total flavones was determined by UV -spectrophotometry an d Basket -Stirring Method.Results Using total flavones as the index,T 50 was 29.57min and T d was 41.28min before wall break-ing,and 41.27min and 54.26min respectively after wall breaking,the dif ference being very significant.Conclusion The dissolution rate of the total fla vones from Pollen Typhae is faster after wall -breaking treatment than th at before treat-ment.
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AIM: To optimize the extraction process of the total flavonoids of Pollen typhae. METHODS: The optimum extraction process was selected with the orthogonal design (L 27 (3 13 )), using the contents of typhaneoside and isorhamnetin 3 O neohesperidoside as the evaluating criteria. RESULTS: The significant effects of alcohol concentration, extraction time and extraction times on the extraction of the total flavonoids of Pollen typhae were discovered. CONCLUSION: The optimum extraction process was as follows: Pollen typhae was extracted with 40% alcohol for two times, each with the solvent volume 14 times of the weight of the raw materials and extraction time for 3 h.
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Objective: To study the extraction process of the total flavones of Pollen Typhae ( Typha angustifolia L.). Methods: The optimum extraction process was selected with the orthogonal design. The content of total flavones was determined by UV spectrophotometry. Results: The significant effects of alcohol concentration, extraction time and extraction times on the extraction of total flavones were discovered. Conclusion: The optimum extraction process was as follows: Pollen Typhae was extracted with 70% alcohol for 3 times, with the solvent volume 8, 6, 6 times amount and extraction time 60, 60, 30 minutes in turn.
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AIM: To investigate the influence of microwave-assisted extraction on flavone contents of Pollen Tyhae micropowder. METHODS: Ultraviolet spectrophotometer and HPLC were applied to analyze Pollen Tyhae micropowder, total flavon and isorhamnetin-3-O-neohespridoside were adopted as the marker, respectively. RESULTS: Appropriate conditions of microwave-assisted extraction included: extraction time of 8min, ethanol concentration of 70%, Solid/liquid ratio of 1∶18 (g?mL -1) and the power of microwave oven of 540w. CONCLUSION: Compared with normal reflux method, microwave-assisted extraction of Pollen Typhae micropowder is more useful and can improve the extraction rate the reduce the extracting time.
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Objective: To study the quality standard for Puhuang Dispensing Granule (Pollen Typhae). Methods : Puhuang Dispensing Granule was identified by TLC and isorhamnetin-3-O-neohesperidoside and typhaneoside in Puhuang Dispensing Granule were determined by HPLC. Results : The linear relationship was at the range of 0.4～1.21?g for isorhamnetin-3-O-neohesperidoside and 0.42～1.25?g for typhaneoside, respectively. The average recovery was 98.88% for isorhamnetin-3-O-neohesperidoside and 99.68% for typhaneoside, respectively. Conclusion : The method is available with a good reproducibility and can control the quailty of Puhuang Dispensing Granule.