Effects and mechanisms of total flavones of Abelmoschus manihot in attenuating diabetic tubulopathy by targeting endoplasmic reticulum stress-induced cell apoptosis / 中国中药杂志

Renal tubular injury in patients with diabetic kidney disease(DKD) may be accompanied by glomerular and microvascular diseases. It plays a critical role in the progression of renal damage in DKD, and is now known as diabetic tubulopathy(DT). To explore the multi-targeted therapeutic effects and pharmacological mechanisms in vivo of total flavones of Abelmoschus manihot(TFA), an extract from traditional Chinese medicine for treating kidney disease, in attenuating DT, the authors randomly divided all rats into four groups: a normal control group(normal group), a DT model group(model group), a DT model+TFA-treated group(TFA group) and a DT model+rosiglitazone(ROS)-treated group(ROS group). The DT rat model was established based on the DKD rat model by means of integrated measures. After successful modeling, the rats in the four groups were continuously given double-distilled water, TFA suspension, and ROS suspension, respectively by gavage every day. After 6 weeks of treatment, all rats were sacrificed, and the samples of their urine, blood, and kidneys were collected. The effects of TFA and ROS on various indicators related to urine and blood biochemistry, renal tubular injury, renal tubular epithelial cell apoptosis and endoplasmic reticulum stress(ERS), as well as the activation of the protein kinase R-like endoplasmic reticulum kinase(PERK)-eukaryotic translation initiation factor 2α(eIF2α)-activating transcription factor 4(ATF4)-C/EBP homologous protein(CHOP) signaling pathway in the kidney of the DT model rats were investigated. The results indicated that hypertrophy of renal tubular epithelial cells, renal tubular hyperplasia and occlusion, as well as interstitial extracellular matrix and collagen deposition occurred in the DT model rats. Moreover, significant changes were found in the expression degree and the protein expression level of renal tubular injury markers. In addition, there was an abnormal increase in tubular urine proteins. After TFA or ROS treatment, urine protein, the characteristics of renal tubular injury, renal tubular epithelial cell apoptosis and ERS, as well as the activation of the PERK-eIF2α-ATF4-CHOP signaling pathway in the kidney of the DT model rats were improved to varying degrees. Therein, TFA was superior to ROS in affecting the pathological changes in renal tubule/interstitium. In short, with the DT model rats, this study demonstrated that TFA could attenuate DT by multiple targets through inhibiting renal tubular ERS-induced cell apoptosis in vivo, and its effect and mechanism were related to suppressing the activation of the PERK-eIF2α-ATF4-CHOP signaling pathway in the kidney. These findings provided preliminary pharmacological evidence for the application of TFA in the clinical treatment of DT.

Effects and mechanisms of total flavones of Abelmoschus manihot in improving insulin resistance and podocyte epithelial-mesenchymal transition in diabetic kidney disease based on IRS1/PI3K/Akt pathway / 中国中药杂志

This study aimed to explore the effects and mechanisms of total flavones of Abelmoschus manihot(TFA), the extracts from traditional Chinese medicine indicated for kidney diseases, on insulin resistance(IR) and podocyte epithelial-mesenchymal transition(EMT) in diabetic kidney disease(DKD), and further to reveal the scientific connotation. Thirty-two rats were randomly divided into a normal group, a model group, a TFA group, and a rosiglitazone(ROS) group. The modified DKD model was induced in rats by methods including high-fat diet feeding, unilateral nephrectomy, and streptozotocin(STZ) intraperitoneal injection. After modeling, the rats in the four groups were given double-distilled water, TFA suspension, and ROS suspension correspondingly by gavage every day. At the end of the 8th week of drug administration, all rats were sacrificed, and the samples of urine, blood, and kidney tissues were collected. The parameters and indicators related to IR and podocyte EMT in the DKD model rats were examined and observed, including the general condition, body weight(BW) and kidney weight(KW), the biochemical parameters and IR indicators, the protein expression levels of the key signaling molecules and structural molecules of slit diaphragm in the renal insulin receptor substrate(IRS) 1/phosphatidylinositol 3-kinase(PI3K)/serine-threonine kinase(Akt) pathway, foot process form and glomerular basement membrane(GBM) thickness, the expression of the marked molecules and structural molecules of slit diaphragm in podocyte EMT, and glomerular histomorphological characteristics. The results showed that for the DKD model rats, both TFA and ROS could improve the general condition, some biochemical parameters, renal appearance, and KW. The ameliorative effects of TFA and ROS were equivalent on BW, urinary albumin(UAlb)/urinary creatinine(UCr), serum creatinine(Scr), triglyceride(TG), and KW. Secondly, they could both improve IR indicators, and ROS was superior to TFA in improving fast insulin(FIN) and homeostasis model assessment of insulin resistance(HOMA-IR). Thirdly, they could both improve the protein expression levels of the key signaling molecules in the IRS1/PI3K/Akt pathway and glomerulosclerosis in varying degrees, and their ameliorative effects were similar. Finally, both could improve podocyte injury and EMT, and TFA was superior to ROS. In conclusion, this study suggested that podocyte EMT and glomerulosclerosis could be induced by IR and the decreased activation of the IRS1/PI3K/Akt pathway in the kidney in DKD. Similar to ROS, the effects of TFA in inhibiting podocyte EMT in DKD were related to inducing the activation of the IRS1/PI3K/Akt pathway and improving IR, which could be one of the scientific connotations of TFA against DKD. This study provides preliminary pharmacological evidence for the development and application of TFA in the field of diabetic complications.

Effects and mechanisms of total flavones of Abelmoschus manihot in inhibiting podocyte necroptosis and renal fibrosis in diabetic kidney disease / 中国中药杂志

Previous studies have shown that high blood glucose-induced chronic microinflammation can cause inflammatory podocyte injury in patients with diabetic kidney disease(DKD). Therein, necroptosis is a new form of podocyte death that is closely associated with renal fibrosis(RF). To explore the effects and mechanisms in vivo of total flavones of Abelmoschus manihot(TFA), an extract from traditional Chinese herbal medicine Abelmoschus manihot for treating kidney diseases, on podocyte necroptosis and RF in DKD, and to further reveal its scientific connotation with multi-pathway and multi-target, the authors randomly divided all rats into four groups: a namely normal group, a model group, a TFA group and a rapamycin(RAP) group. After the modified DKD rat models were successfully established, four group rats were given double-distilled water, TFA suspension and RAP suspension, respectively by gavage every day. At the end of the 4th week of drug treatment, all rats were sacrificed, and the samples of their urine, blood and kidneys were collected. And then, the various indicators related to podocyte necroptosis and RF in the DKD model rats were observed, detected and analyzed, respectively. The results indicated that, general condition, body weight(BW), serum creatinine(Scr), urinary albumin(UAlb), and kidney hypertrophy index(KHI) in these modified DKD model rats were both improved by TFA and RAP. Indicators of RF, including glomerular histomorphological characteristics, fibronectin(FN) and collagen type Ⅰ(collagen Ⅰ) staining extent in glomeruli, as well as the protein expression levels of FN, collagen Ⅰ, transforming growth factor-β1(TGF-β1) and Smad2/3 in the kidneys were improved respectively by TFA and RAP. Podocyte damage, including foot process form and the protein expression levels of podocin and CD2AP in the kidneys was improved by TFA and RAP. In addition, tumor necrosis factor-α(TNF-α)-mediated podocyte necroptosis in the kidneys, including the morphological characteristics of podocyte necroptosis, the extent and levels of the protein expression of TNF-α and phosphorylated mixed lineage kinase domain like pseudokinase(p-MLKL) was improved respectively by TFA and RAP. Among them, RAP had the better effect on p-MLKL. More importantly, the activation of the receptor interacting serine/threonine protein kinase 1(RIPK1)/RIPK3/MLKL signaling axis in the kidneys, including the expression levels of its key signaling molecules, such as phosphorylated receptor interacting serine/threonine protein kinase 1(p-RIPK1), p-RIPK3, p-MLKL and cysteinyl aspartate specific proteinase-8(caspase-8) was improved respectively by TFA and RAP. Among them, the effect of TFA on p-RIPK1 was superior. On the whole, in this study, the authors demonstrated that TFA alleviates podocyte necroptosis and RF in DKD through inhibiting the activation of the TNF-α-mediated RIPK1/RIPK3/MLKL signaling axis in diabetic kidneys. The authors' findings provide new pharmacological evidence to reveal the scientific connotation of TFA in treating RF in DKD in more depth.

Effects of total flavonoids of Bidens pilosa on insulin resistance in HepG 2 cells / 中国药房

OBJECTI VE To explore the effects of total flavonoids of Bidens polisa L.（TFB）on insulin resistance （IR）of HepG2 cells. METHODS B. polisa L. was refluxed and extracted with 80％ ethanol to obtain TFB. Palmitic acid was used to induce IR mode of HepG 2 cells in vitro . The effects of low-concentration ，medium-concentration and high-concentration （20，40， 80 mg/L） of TFB on the consumption of glucose were investigated. Using metformin as positive control ，the effects of low-concentration，medium-concentration and high-concentration （20，40，80 mg/L）of TFB on the protein expression of insulin receptor substrate- 1（IRS-1），c-Jun N-terminal kinase （JNK）and protein kinase C （PKC）were investigated. Molecular docking technology was used to explore the interaction between eight main active components of TFB such as quercetin ，quercitrin and IRS-1，JNK and PKC proteins. RESULTS The glucose consumption of TFB low-concentration ，medium-concentration and high-concentration groups were increased significantly （P＜0.05 or P＜0.01）. Compared with normal group ，the expression of IRS-1 and JNK protein in the model group decreased significantly ，and the expression of PKC protein increased significantly （P＜ 0.01）. Compared with model group ，the protein expression of IRS- 1 and JNK could up-regulated while the protein expression of PKC down-regulated in TFB low-concentration ，medium-concentration and high-concentration groups and metformin positive control group （P＜0.05 or P＜0.01）. The score of molecular docking energy between maritimetin in TFB and IRS- 1 protein was －7.9 kcal/mol（1 kcal＝4.816 kJ）. The scores of molecular docking energy of maritimetin ，rutin and JNK protein were －9.3 kcal/mol. The score of molecular docking energy between quercitrin and PKC protein was －4.9 kcal/mol. Interactions between components and proteins included forming hydrogen bonds ，hydrophobic bonds and so on. CONCLUSIONS TFB can significantly improve IR of HepG 2 cells，the mechanism of which may be related to the regulation of protein expression of IRS ，JNK and PKC. Maritimetin，rutin and quercitrin may be potential active ingredients for improving IR.

Content determination of total flavones and polysaccharides in Fangshu Qingre mixture by UV-Vis spectrophotometry / 药学实践杂志

Objective To establish a method for content determination of total flavones and polysaccharides in Fangshu Qingre mixture by UV-Vis Spectrophotometry. Methods The contents of total flavones and polysaccharides in Fangshu Qingre mixture were determined by UV-Vis spectroscopy with rutin and anhydrous glucose as reference substance, and the wavelength was set at 508 nm and 487 nm. Results The contents were from 0.00 to 59.20 μg/ml for total flavones and from 10.92 to 109.20 μg/ml for total polysaccharides in Fangshu Qingre mixture. The recoveries of total flavones and total polysaccharides were 104.4% and 104.8% respectively. Conclusion The method of using ultraviolet spectroscopy was simple, reproducible, accurate and reliable, which could be preferably used as the method for content determination of total flavones and polysaccharides in Fangshu Qingre mixture.

Effects of lactagogue of four total alkaloids of barley malt via dopamine D2 receptor in postpartum hypogalactia rat / 中草药

Objective: To explore the regulatory effect of malt alkaloid on prolactin (PRL) secretion in the model rat with postpartum hypogalactia induced by bromocriptine based on dopamine D2 receptor, and determine the active fraction of the malt with galactogogue effect. Methods: The postpartum hypogalactia model was established by intragastric administration of bromocriptine mesylate. After the model was successfully established, all groups were given corresponding drug treatment. The concentration of serum PRL, estradiol (E2) and progesterone (P) in each group was detected by ELISA kits. HE staining was used to observe the pathologic changes of breast tissue. RT-PCR was used to determine the mRNA levels of prolactin receptor (PRLR) and dopamine D2 receptor (DRD2) in pituitary gland of rats. Results: Compared with the control group, the levels of serum PRL, P, and E2 were significantly decreased in the model group as well as the mRNA expression of the pituitary PRL cells. But the mRNA expression of the pituitary DRD2 in the model group was significantly increased compared with the control group. Compared with the model group, the malt total alkaloid significantly increased the volume of mammary lobule and dilated the duct. There was a lot of milk in the duct and acinar in the malt total alkaloid group. Besides, the total alkaloids increased the concentration of serum PRL, P, and E2 and the mRNA expression of the pituitary PRL cells, and decreased the mRNA expression of the pituitary DRD2. Conclusion: The primary the active fraction of malt for galactogogue action is total alkaloids, and its mechanism may be related to promoting PRL secretion, increasing serum PRL receptor level and decreasing the mRNA expression of dopamine D2 receptor.

Effects of total flavones of rhododendra on contractility of rat myocardial cells and its mechanism / 安徽医科大学学报

Objective To investigate the total flavones of rhododendra(TFR) on contractility of rat myocardial cells and its possible mechanism. Methods The contraction amplitude and contraction frequency of primary cultured rat myocardial cells were observed by image analysis system. The intracellular free Ca2 + content was measured by calcium ion imaging system. Results 10 and 100 nmol /L hU Ⅱ significantly accelerated the contraction frequency of myocardial cells,and 10 nmol /L hU Ⅱ increased the contraction amplitude of myocardial cells,but 100 nmol /L hU Ⅱ reduced the contraction amplitude of myocardial cells. TFR 300 mg /L significantly slowed the contraction frequency of rat myocardial cells and increased the contraction amplitude. TFR in the range of 33. 3 ～ 300 mg /L could significantly inhibit the increase of contraction frequency,the decrease of contraction amplitude and the increase of intracellular free Ca2 + content induced by 100 nmol /L hU Ⅱ. Conclusion TFR can slow down the contraction frequency of myocardial cells and increase its contractility,which may be related to the decrease of free Ca2 + content in myocardial cells.

Study on Protective Effect of Total Flavonoids from the Leaves of Mongolian Medicine Choerospondias axillaris on Myocar- dial Ischemia Reperfusion Injury Model Rats / 中国药房

OBJECTIVE： To investigate the protective effect of total flavonoids from the leaves of Choerospondias axillaris （TFLC） on myocardial ischemia reperfusion injury （MIRI） model rats. METHODS： Male SD rats were randomly divided into sham operation group， model group， positive control group （verapamil， 0.02 g/kg）， TFLC low-dose and high-dose groups （0.1， 0.4 g/kg）， with 10 rats in each group. Administration groups were intragastrically given relevant medicine （2 mL/100 g）； sham operation group and model group were given constant volume of normal saline intragastrically， once a day， for consecutive 7 d. After last medication， MIRI model was induced by modified ligation method. The times and duration of ventricular tachycardia （VT） and ventricular fibrillation （VF） in rats were recorded with biological function experiment system during reperfusion period.The activity of CK and contents of TNF-α， IL-6， NF-κB and NO in serum were determined by ELISA double antibody clip art assay. The morphological characteristics of myocardial tissue was observed by HE staining. The myocardial infarction scope （i.e. the ratio of myocardial tissue mass to ventricular mass） was measured by TTC method. RESULTS： Compared with sham operation group， the times and duration of VT and VF were increased or prolonged significantly in model group； CK activity， serum contents of TNF-α， IL-6 and NF-κB were enhanced or increased significantly， while NO content was decreased significantly （P＜0.05 or P＜0.01）. Obvious myocardial infarction focus， serious cell structure damage， disorderly muscle fibers arrangement， cell nucleus pyknosis and accompanied inflammatory cell infiltration were all observed in cardiac tissue； the mass of infarcted myocardial tissue and ventricular as well as the scope of myocardial infarction increased significantly （P＜0.01）. Compared with model group， the times and duration of VT and VF were decreased or shortened significantly in administration groups； CK activity， serum contents of TNF-α， IL-6， NF-κB were decreased significantly， while NO content was increased significantly （P＜0.05 or P＜0.01）. The above symptoms of myocardial injury were improved；the mass of infarcted myocardial tissue and ventricular as well as the scope of myocardial infarction was significantly reduced （P＜0.05 or P＜0.01）. CONCLUSIONS： TFLC can relieve MIRI-induced ischemic arrhythmia and myocardial damage， reduce the release of inflammatory factors， promote the recovery of myocardial and endothelial cell function， reduce the scope of myocardial infarction and has a certain protective effect.

Effect of Rosae Chinensis Flos Total Flavones on Focal Cerebral Ischemia-reperfusion in Rats / 中国实验方剂学杂志

Objective: To study the effect of Rosae Chinensis Flos total flavones(RCTF) on the focal cerebral ischemia-reperfusion model in rats, in order to preliminarily explore the mechanism of action. Method: Rats were randomly divided into sham-operated group, model group, large, medium, and low-dose RCTF group(200,100,50 mg ·kg-1) and positive group[Nimodipine group(20 mg ·kg-1) and Naoluotong group (500 mg ·kg-1)]. After 7 days of continuous administration, 1 hour later after the last administration, the middle cerebral artery middle cerebral artery occlusion (MCAO) model was duplicated. After 2 hours of modeling, perfusion was performed for 22 hours. Mortality and neurological deficits were scored. Serum S-100β was detected; brain tissue malondialdehyde(MDA), superoxide dismutase (SOD), nitric oxide (NO), nitric oxide synthase (NOS), tumour necrosis factor-α(TNF-α), interleukin-1β(IL-1β), intercellular adhesion molecule-1(ICAM-1), adenosine triphosphate (ATP)ase were measured. The brain tissue morphological changes were observed. Result: The rat model of focal cerebral ischemia and reperfusion was successfully replicated. Compared with the model group, RCTF in large, medium, and low-dose RCTF group significantly decreased the score of neurological deficit in rats (Pβ in serum (PPP+K+-ATPase, Mg2+-ATPase, and Ca2+ in brain tissue (Pα content, IL-1β, ICAM-1 content in brain tissue (PPConclusion: RCTF have a protective effect on cerebral ischemia-reperfusion injury in rats. The mechanism may be related to the resistance of anti-free radicals, the reduction of inflammation in brain tissue and the improvement of brain energy metabolism after cerebral ischemia reperfusion injury.

Content Determination of Total Flavones and Polysaccharides in Hedysari Radix in Different Producing Areas in Gansu Province / 中国中医药信息杂志

Objective To establish a method for content determination of total flavones and polysaccharides in Hedysari Radix in different producing areas in Gansu Province.Methods The contents of total flavones and polysaccharides in Hedysari Radix in different producing areas in Gansu Province were determined by ultraviolet spectroscopy with calycosin-7-glucoside and glucose as reference substance, and the wavelength was set at 260 nm and 484 nm.Results The contents were from 2.82 mg/g to 6.79 mg/g for total flavones and from 106.14 mg/g to 746.40 mg/g for total polysaccharides in Hedysari Radix in different producing areas in Gansu Province. The recoveries of total flavones and total polysaccharides were 97.96％ and 102.90％, respectively.Conclusion There was difference in contents of total flavones and polysaccharides of Hedysari Radix in different producing areas in Gansu Province, and the method of using ultraviolet spectroscopy is simple, reproducible, accurate and reliable, which can be preferably used as the method for content determination of total flavones and polysaccharides in Hedysari Radix.

Comparison of the quantitative analytical methods of total flavones in three quality inspection regulations / 国际药学研究杂志

Objective To compare the difference of the results of the total flavones acquired by three kinds of quantitative an-alytical methods in the People's Republic of China Pharmacopoeia(method A),Health Food Function Components and Health Indica-tors Inspection Specifications(method B),and Health Food Inspection and Evaluation of Technical Specifications(method C)respec-tively. Methods Twenty-one drug and food homologies,medicines and other samples were detected by using the above mentioned methods respectively. Comparisons of the results were established. Results The measured values by method A were the highest,that of method C were the lowest. Moreover,the determination values of method A were higher than those by the methed B(1.4 ± 0.12) times,and the method C(2.1±0.31)times. Conclusion There are significant differences among the results of the total flavonoids de-terminated by the 3 methods in the same sample and the numerical relationship will be helpful for the quantity control of homologous food and drug.

Protective effects of total flavones from Xueli Formula on LPS-stimulated RAW264.7 cell inflammation / 中成药

AIM To observe the anti-inflammatory effects of total flavones from Xueli Formula (Violae Herba,Serissa japonica,Plantaginis Herba and Salvia plebeia,TFXL) on LPS-stimulated RAW264.7 macrophages.METHODS The effects of different concentrations of TFXL on RAW264.7 cell viability were examined by MTT assay.The NO kit assay was adopted to detect the NO release amount of TFXL on LPS-stimulated RAW264.7 cells.The secretions of tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6) and interleukin-10 (IL-10) were tested by enzyme linked immunosorbent assay (ELISA).Reverse transcriptase-polymerase chain reaction (RTPCR) was used to determine the expressions of inducible nitric oxide synthase (iNOS),TNF-α,IL6 and IL10 mRNA.The protein expressions of IκB-α and p65 were tested by Western blot.RESULTS Compared with the LPS model group,TFXL could significantly reduce the secretions of NO,TNF-α and IL-6;increase the secretion of IL-10;inhibit the expressions of iNOS,TNF-α and IL6 mRNA;promote the expression of IL10 mRNA;inhibit the phosphorylations of IκB-α and p65.The TFXL high-dose group could inhibit the degradation of IκB-α.CONCLUSION This study preliminarily proves the protective effects of TFXL on LPS-stimulated RAW264.7 cell inflammation,whose action mechanism may be related to NF-κB signal pathway.

Extraction Optimization of Qishan Sugar-free Granule by Orthogonal Design / 中国药师

Objective:To study the best extraction technology of Qishan sugar-free granule. Methods:The effects of boiling dura-tion, boiling times and water addition ratio on the extraction process were investigated by using L9 (34 ) orthogonal design, and the con-tents of total flavonoids, baicalin and berberine were taken as the indices. Results:The optimum conditions were as follows:10-fold a-mount of water for 2 cycles, and every extracting time was 1 hour. Conclusion: The best extraction technology of Qishan sugar-free granule is obtained, and the process is stable and feasible, and the contents of active components are high.

Effect of Dracocephalum moldavica total flavones on expression of TGF-β1/Smad signaling pathway and matrix metalloproteinase of atherosclerosis ApoE-/- mice / 中国中药杂志

To investigate the effect and mechanism of Dracocephalum moldovica total flavones (TFDM) on the formation of atherosclerosis ApoE-/- mice induced by high fat diet. A total of 40 SPF 8-week-old male ApoE-/- mice were fed with high fat diet and randomly divided into 5 groups. TFDM high, medium, low-dose group were given 21, 42, 84 mg•kg⁻¹•d⁻¹ by gavage; Simvastatin group was fed with simvastatin 3.5 mg•kg⁻¹•d⁻¹; and model group was given the same dose of normal saline. The other eight male C57BL/6J mice of the same genetic background and age were set up as control group and fed with common diet. All of the groups were intragastrically intervened for 12 weeks. The aortic pathologic changes were observed with HE; qRT-PCR was adopted to detect TGF-β1, Smad2, Smad3, MMP-2 and MMP-9 gene levels in tissues. Compared with model group, HE staining in TFDM group showed obvious relief of aortic atherosclerotic tissue injury; each TFDM group showed inhibition in mRNA expressions of TGF-β1, Smad2, Smad3, MMP-2 and MMP-9. This suggests that TFDM can inhibit atherosclerosis formation, which may be related to the intervention of TGF-β1/Smads signal transduction.

Mechanisms of total flavones of Epimedium on oxidative stress induced by myocardial ischemia/reperfusion injury in rats / 中国中药杂志

To investigate the effects and mechanisms of total flavones of Epimedium (TFE) on oxidative stress induced by myocardial ischemia/reperfusion injury in rats, forty male SD rats were randomly divided into sham operated group, model group, diltiazem group and flavonoids of Epimedium low and high doses groups with 8 rats in each. Myocardial ischemia/reperfusion injury model was induced by ligaturing the left anterior descending artery for 30 min followed reperfusion for 4 h after TFE was taken by intragastric administration for 4 days. The degree of myocardial infarct was observed by N-BT staining. The concentrations of MDA and activities of SOD and T-AOC in cardiac tissue were measured by colorimetry. Serum TnI concentrations were checked by ELISA. HE stain was used to observe myocardium structure under light microscope. Expressions of SIRT1 and Nrf2 in cardiac tissue were evaluated by immunohistochemistry method and Western blot, respectively. Compared with the model group, the degree of myocardial infarct, MDA concentration in cardiac tissue and the levels of TnI in serum significantly decreased in the diltiazem group and flavonoids of Epimedium low and high doses groups (P<0.05 or P<0.01); flavonoids of Epimedium low and high doses groups and the diltiazem group also showed improvements in myocardium structure under ischemia/reperfusion injury. TFE significantly increased the activity of SOD and T-AOC and the expression of SIRT1 and Nrf2 in cardiac tissue when compared with the model group (P<0.05 or P<0.01). Therefore, TFE can increase anti-peroxidant capacity of myocardium tissue by using intrinsically anti-oxidant signaling pathway of SIRT1 and Nrf2, which can inhibit irreversible damage of cardiomyocytes in myocardial ischemia/reperfusion injury and protect normal function of cardiac tissue.

Hepatoprotective Effect of Total Flavones of Artemisia capillaris Thunb. on Acute Hepatic Injury in Rats / 医药导报

Objective To investigate the protective effect of total flavones of Artemisia capillaris Thunb.on acute hepat-ic injury in rats. Methods Rats were randomly divided into blank control group,model control group, Yinzhihuang group,and groups of total flavones of Artemisia capillaris Thunb.(low,medium and high dose) in terms of 7-day different treatments.All rats except those in the blank control group were administrated with D-galactosamine hydrochloride ( 500 mg?g-1 , ip ) once at the sixth day.Then,concentrations of ALT and AST were detected 48 h later,and the liver samples were collected from each group for pathological examination. Results The serum ALT and AST in high-dose group of total flavones of Artemisia capillaris Thunb. was [(189.2±112.9) and (231.7±149.9) U?L-1],respectively,significantly lower than those in model control group ALT [(391.9±181.3) U?L-1] and AST [(403.9±133.8) U?L-1].Fragmented necrosis,fatty degeneration,inflammatory cells infil-tration and acidophilic degeneration of hepatic cells were improved to varying degrees in groups of total flavones of Artemisia capil-laris Thunb.compared with model control group.Fragmented necrosis of liver cells and steatosis occurred in 20 and 19 rats,respec-tively,in the model control group,while those appeared in 1 and 2 rats,respectively,in high-dose group of total flavones of Artemi-sia capillaris Thunb.. Conclusion Total flavones of Artemisia capillaris Thunb. are effective in protecting D-galactosamine hydrochloride-induced acute hepatic injury in rats.

Effect and mechanism of total flavones of oldenlendia diffusa willd on epithelial-mesenchymal transition of cell line MHCC97-H induced by TGF-β1 / 西安交通大学学报(医学版)

ABSTRACT:Objective To investigate the effects of total flavones of oldenlandia diffusa (FOD)on epithelial-mesenchymal transition in hepatocellular cancer cell line MHCC97-H.Methods TGF-β1 induced EMT in routinely cultured liver cancer cell line MHCC97-H;then MHCC97-H cell was divided into 5 groups:normal control group, TGF-β1 group,TGF-β1 + FOD group,TGF-β1 + 5-FU group,and TGF-β1 + FOD + 5-FU group.After 48 h of treatment,the invasion ability of MHCC97-H cell was detected by Transwell;the proteins of E-cadherin and vimentin were determined by Western blot.Results Compared with the normal form of MHCC97-H cell line,the cell had obvious long fusiform after TGF-β1 induction,and the invasion ability enhanced (P = 0.02 ).But after treatment,the invasion ability of MHCC97-H cell decreased in FOD group and 5-FU group compared with that in TGF-β1 group (P = 0.03,P = 0.02 ),and decreased more significantly in FOD + 5-FU group (P = 0.01 ).The expression of E-cadherin at the protein level decreased significantly (P = 0.01 )in TGF-β1 group,which was abolished in FOD group (P =0.03 )and 5-FU group (P = 0.02 ).The expression of vimentin at the protein level increased significantly (P =0.01)in TGF-β1 group,which was abolished in FOD group (P =0.04)and 5-FU group (P =0.03)and more obviously in FOD+5-FU group (P =0.01).Conclusion FOD can reverse the invasion of MHCC97-H cells in EMT induced by TGF-β1 through decreasing the expression of E-cadherin protein and inhibiting the epithelial-mesenchymal transition of MHCC97-H cell.

Effect of Rhizoma Drynariae Total Flavones on Expression of Bone Morphogenetic Protein 2 and Transforming Growth Factorβ1 in Distraction Osteogenesis / 广州中医药大学学报

Objective To investigate the effect of total flavones of Rhizoma Drynariae on femur distraction osteogenesis in the rabbits. Methods Thirty-two healthy rabbits were randomly divided into treatment group and control group, 16 rabbits in each group. The femoral fracture was treated with unilateral femoral distraction and was fixed with a self-made distraction instrument. After 7-day intermittent period, the fractured femur was distracted at a rate of 1 mm/d, twice a day for 10 continuous days. The treatment group was fed with total flavones of Rhizoma Drynariae from the first post-operative day to the end of the experiment. And then all of the animals were sacrificed after fixation for 28 days. The bone specimens were used for histological observation and immunohistochemical detection. Results The area of mature bone in the newborn bone tissue of the treatment group was increased, and osteoblasts number and the percentage of trabecular bone area were significantly higher than those of the control group . The bone morphogenetic protein-2 (BMP-2) and transforming growth factor-β1 (TGF-β1) were stained brown deeply, the staining degree being stronger than that of the control group. Conclusion Rhizoma Drynariae total flavones can effectively accelerate the formation and maturation of newborn bone tissue during bone distraction.

Protective effect of total flavones of buckwheat flowers and leaves on myocardial fibrosis of type 2 diabetic rats and its mechanism / 吉林大学学报(医学版)

Objective: To study the protective effect of the total flavones of buckwheat flowers and leaves (TFBFL)on the myocardial injury of the type 2 diabetic rats,and to clarify its mechanism.Methods:The diabetic rat models were established by intraperitoneally injecting streptozotocin (STZ)and taking high-fat diet.The model rats were divided into diabetic model (DM)group and TFBFL treatment (TFBFL)group,while normal control group was set up, 10 rats per group.The rats in TFBFL group were administered with 200 mL· kg-1 · d-1 TFBFL,while the rats in other two groups were given with normal water (10 mL·kg-1 ·d-1 )instead.The rats in various groups was administered once a day for 8 weeks.The body weight (BW),heart weight index (HWI), level of fasting blood glucose (FBG)and cardiac function indexes (HR,LVSP,LVEDP and ± dp/dtmax )of the rats in various groups were measured.The myocardial tissue morphology of the rats in various groups was evaluated by electron microscope and the collagen levels in myocardium tissue of the rats in various groups were observed by Masson staining.The expression levels of TGF-β1 protein in myocardium tissue of the rats in various groups were detected by Western blotting method.Results:Compared with normal control group,the HWI,FBG and LVEDP levels of the rats in DM group were increased (P 0.05).The Masson staining results showed the morphology of myocardium tissue of the rats in TFBFL group was improved and the collagen level was decreased. The Western blotting results suggested that the TGF-β1 protein expression level in myocardium tissue of the rats in DM group was significantly increased compared with normal control group (P < 0.05),and the TGF-β1 protein expression level of the rats in TFBFL group was decreased compared with DM group (P <0.05).Conclusion:TFBFL exerts a certain protective effect on the myocardial fibrosis of diabetic rats through inhibiting the TGF-β1 expression in myocardium tissue.

Glucolipid metabolism and antioxidant effect of the total flavones of propolis in GK rats / 中国药学杂志

OBJECTIVE: To study on glucolipid metabolism effect of the total flavones of propolis (TFP) in Goto-Kakisaki rats (GK Rats). METHODS: Fifty GK rats were randomly divided into 5 groups by blood glucose, the model group, positive drug group (2 000 mg · kg-1) and the high (240 mg · kg-1) dose of the TFP group, the medium (120 mg · kg-1) doses of the TFP group, low (60 mg · kg-1) doses of the TFP group. The GK rats in TFP groups were given with TFP by intragastric administration, continuously treated for 8 weeks, the control group were given equal sodium carboxymethylcellulose(CMC-Na) and positive drug group were given (2 000 mg · kg-1) dose of Xiaoke pills by intragastric administration. The glucose (Glu), total cholesterol (TC), triglyceride (TG), high density lipoprotein(HDL-C), low density lipoprotein(LDL-C), glycated hemoglobin(GHb), superoxide dismutase (SOD), ma-londialdehyde(MDA), glutathione(GSH), serum insulin(lNS), C-peptide(C-P), tumor necrosis factor-α(TNF-α) free fatty acid (FFA), nitric oxide(NO), and hepatic glycogen were measured at the end of the experiment. RESULTS: Compared with the control group, the levels of GLU, TG, TC, LDL-C, GHb, MDA and TNF-α were decreased significantly (P 0.05) in the low-dose group. The levels of SOD, hepatic glycogen, NO and C-P were increased significantly(P < 0.05 or P < 0.01), the levels of GLU, MDA, TG and GHb were decreased significantly (P < 0.05 or P < 0.01 or P < 0.001) in Xiaoke pills group. CONCLUSION: TFP can significantly decrease the level of blood glucose, significantly improve the glucose and lipid metabolism and inhibit insulin resistance in deficiency of GK rats.