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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.
Subject(s)
Rats , Animals , Diabetic Nephropathies/drug therapy , Proto-Oncogene Proteins c-akt/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Abelmoschus/chemistry , Podocytes , Rats, Sprague-Dawley , Epithelial-Mesenchymal Transition , Flavones/pharmacology , Insulin Resistance , Reactive Oxygen Species , Diabetes MellitusABSTRACT
Angelicae Sinensis Radix (Danggui) and Ligusticum Chuanxiong Rhizoma (Chuan Xiong) herb-pair (DC) have been frequently used in Traditional Chinese medicine (TCM) prescriptions for hundreds of years to prevent vascular diseases and alleviate pain. However, the mechanism of DC herb-pair in the prevention of liver fibrosis development was still unclear. In the present study, the effects and mechanisms of DC herb-pair on liver fibrosis were examined using network pharmacology and mouse fibrotic model. Based on the network pharmacological analysis of 13 bioactive ingredients found in DC, a total of 46 targets and 71 pathways related to anti-fibrosis effects were obtained, which was associated with mitogen-activated protein kinase (MAPK) signal pathway, hepatic inflammation and fibrotic response. Furthermore, this hypothesis was verified using carbon tetrachloride (CCl
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Objective: To investigate the toxicity and mechanisms of celastrol (CEL) on human biliary epithelial cells. Methods: The effects of CEL on cell morphology and cell viability changes were observed by CCK-8 experiment and microscope. Cell scratch experiment was used to detect the effect of CEL on cell migration. The effects of CEL on cell cycle and cell apoptosis were detected by flow cytometry. The mRNA and protein expression of apoptosis-related genes Caspase-3, Bax and Bcl-2 were detected by qRT-PCR and Western blotting. Results: CEL inhibited cell proliferation and changed cell morphology at 400-2 000 nmol/L. At 200-800 nmol/L, cell migration was inhibited. At 800-1 200 nmol/L, G0/G1 phase was arrested. At 400-1 200 nmol/L, cell apoptosis was induced and the expression of apoptosis-related genes was increased. Conclusion: CEL showed cholangiocyte toxicity through affecting cell viability, cell migration, preventing cell cycle and promoting cell apoptosis of human biliary epithelial cells.
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Shaoyao Gancaotang, first seen in Treatise on Febrile and Miscellaneous Diseases, is composed of Paeoniae Radix Alba and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle in equal proportion.It has the functions of preserving Yin to nourish blood, harmonizing liver and spleen, relieving spasm and pain.This formula is applied in leg and foot spasm and abdominal pain caused by blood deficiency, body fluid consuming and the unmoistened muscles and veins.It has been highly praised and used by medical experts throughout the ages and has extended its application scope.Modern pharmacological studies have shown that Shaoyao Gancaotang has significant effects in antispasmodic, analgesic, antitussive and other areas, and is used to treat spastic diseases, painful diseases, inflammatory diseases and so on.This paper will systematically elaborate the historical evolution, compatibility analysis, pharmacological and pharmacodynamic studies, modern clinical application of Shaoyao Gancaotang, in order to provide theoretical basis and reference for the development of this famous classical formula.
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Objective:To investigate the relation between aquaporins (AQPs) and fecal water content in rats with spinal cord injury. Methods:A total of 48 female Sprague-Dawley rats were divided into control group (n= 24) and spinal cord injury group (SCI group,n = 24). SCI group underwent transection at T8, while the control group was only subjected to laminectomy. Posterior limb function was assessed by Basso-Beattie-Bresnahan (BBB) score before modeling and on the 1st, 3rd, 7th, 14th and 28th day after SCI. Fecal water content was measured before modeling and on the 3rd, 14th and 28th day after SCI. Colon specimens were collected to detect the expression of AQP1, AQP3 and AQP4 by immunohistochemistry on the 3rd, 14th and 28th day after SCI. Results:The BBB score was significantly lower in SCI group than in the control group (t > 69.230,P< 0.001) after SCI, as well as the fecal water content (t > 5.814,P< 0.001). The expression of AQP1, AQP3, and AQP4 in the colon was higher in SCI group than in the control group (|t|> 5.165,P < 0.01) on the 3rd, 14th, and 28th day after SCI. The expression of AQPs negatively correlated with fecal water content (r = -0791~-0.730,P< 0.001). Conclusion:The expression of AQPs in the colon of rats after SCI was up-regulated, which was correlated with excessive water absorption after SCI.
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Objective:To investigate the changes of metabolite concentration in the precentral gyrus (primary motor cortex, M1) in spinal cord injury (SCI) patients. Methods:From December, 2018 to October, 2019, 20 SCI patients and 15 healthy controls were scaned with magnetic resonance spectroscopy to measure the concentrations of N-acetylaspartate (NAA), choline (Cho), creatine (Cr) and myo-inositol (MI) in region of interest (ROI) of left M1. Results:Concentration of MI was more in the patients than in the controls (t = 3.745, P < 0.01). There was no significant difference in the concentrations of NAA, Cho and Cr, as well as the ratios of NAA/Cr, Cho/Cr, Cho/NAA between the patients and the controls (t < 1.431, P > 0.05). Conclusion:There may be hyperplasia of glial cells in M1 of SCI patients, indicating compensatory repair in cerebral motor cortex.
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Vanda falcata (Thunb.) Beer (Orchidaceae), a famous native orchid of China, Japan, and Korea, is known as one of the most beautiful and charming orchid species in the world (Ohwi, 1965; Lawler, 1984; Arditti, 2008). V. falcata is widely cultivated and delights the world with its compact plant shape, elegant white blooms, and sweet coconut-like scent. However, vegetative propagation by division has limited the development of V. falcata because of its inefficiency (Mitsukuri et al., 2009a, 2009b).
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Objective:To study the dry extract rate,determination and transfer rate of maker compounds,fingerprint and others of standard decoction of Chrysanthemi Indici Flos and provide basic data for the preparation of this standard decoction and its dispensing granules by establishing 15 batches of standard decoction of Chrysanthemi Indici Flos from 5 different places. Method:The standard decoction of Chrysanthemi Indici Flos was prepared based on the traditional decoction process,the content of linarin was determined by UPLC-DAD,the transfer rate of this composition was calculated,the fingerprint was drawn,the extract powder was prepared by vacuum drying,and the dry extract rate was calculated. Result:The concentration of linarin in 15 batches of standard decoction of Chrysanthemi Indici Flos was 0.19-0.74 g·L-1,the transfer rate of linarin was 21.95%-66.23%,its average transfer rate was 37.12% with RSD of 11.8%,the pH value was 5.1-5.5,the range of dry extract rate was 24.7%-32.5%,the average dry extract rate was 27.87% with RSD of 2.4%.There were 9 major common peaks in the fingerprint and 2 peaks(No. 2 and No. 9) were confirmed,such as chlorogenic acid and linarin. Conclusion:The preparation method in this research conforms to the traditional decoction method and is stable and feasible.It can be used for the preparation and quality evaluation of standard decoction of Chrysanthemi Indici Flos.
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Objective To compare and analyze the transcriptome of rhizome and leaves of Dioscorea zingiberensis, and excavate the key enzyme genes related to the saponin biosynthetic pathway in D. zingiberensis. Methods The transcriptome of rhizome and leaves of D. zingiberensis were sequenced by Illumina HiSeq2000 high-throughput sequencing technique. According to sequence annotate results to find the differentially expressed genes. Then the key enzyme genes related to the biosynthesis of diosgenin were identified according to the content of saponins in rhizomes and leaves of D. zingiberensis. The expression levels of some candidate genes were analyzed by real-time fluorescence quantitative PCR (qRT-PCR). Results A total of 81 660 Unigenes were gained and 64.33% of them were annotated in NT, NR, Swiss-Prot, KOG, GO, and KEGG databases. Based on their expression and KEGG annotation, totally 227 catalytic enzyme genes of 29 kinds that may participate in D. zingiberensis saponin biosynthetic pathway were screened. The expression pattern of some catalytic enzymes was correlated with the content of saponin. Also were found five D. zingiberensis endophyte genes. Conclusion This experiment obtained candidate key enzyme genes tentatively that involved in the biosynthesis of saponin. Some candidate enzyme genes may participate in the post-modification process of steroidal saponins in D. zingiberensis. In additon, it was found that D. zingiberensis endotrophic bacteria may be involved in the saponin biosynthesis. The results laid a foundation to further elucidate the molecular mechanism of sapogenin synthesis pathway.
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Based on plant species databases, species lists and literature records, general situation of the Chinese medicinal endemic plant (vascular plant) has been systematically summarized, and its quantity and distribution characteristics of Chinese medicinal endemic plants are presented in this paper. The results showed that 3 150 endemic species are Chinese medicinal plants belonging to 785 genera in 153 families, which includes 38 species of 22 genera in 12 families of pteridophyta, 42 species of 14 genera in7 families of gymnosperms, and 3 070 species of 749 genara in 134 families of angiosperms. The top four families involving medicinal endemic species are Asteraceae (218 species), Ranunculaceae (182 species), Labiatae (151 species), and Liliaceae (133 species). The top four provincial administration distributed medicinal endemic species are Sichuan (1 568 species), Yunnan (1 533 species), Guizhou (955 species) and Hubei (930 species).On the regional scale, the most abundant one is the southwest region (2 465 species), followed by the central region (1 226 species) and the northwest region (949 species). Localization characteristics for domestication and artificial cultivation of medicinal endemic species are more prominent due to their narrower and limited distribution areas, indicating it is possible for these species acting as local potential resource for reasonable economic development.
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An HPLC method was established to determine the contents of catalpol, acteoside, rehmaionoside A, rehmaionoside D, leonuride in three part of Rehmanni glutinosa in Beijing No.1 variety R. glutinosa during the growth period, This method, in combination with its HPLC fingerprint was used to evaluate its overall quality characteristics.The results showed that：① the content of main components of R. glutinosa varied in different growth stages ;② there was a great difference of the content of main components between theradial striations and the non-radial striations; ③ the two sections almost have the same content distribution of catalpol, acteoside and rehmaionoside D; ④the content of rehmaionoside A in non-radial striations was higher than that in radial striations,while the content of leonuride in radial striations was higher than that in non-radial striations.; ⑤the HPLC fingerprint of radial striations, non-radial striations and whole root tuber were basically identical, except for the big difference in the content of chemical components. The result of clustering displayed that the radial striations, non-radial striations, and whole root were divided into two groups. In conclusion, there was a significant difference in the quality characteristics of radial striations and non-radial striations of R. glutinosa. This research provides a reference for quality evaluation and geoherbalism of R. glutinosa.