Radix Scrophulariae Extracts Exert Effect on Hyperthyroidism via MST1/Hippo Signaling Pathway / 中国结合医学杂志

OBJECTIVE@#To explore the mechanism of Radix Scrophulariae (RS) extracts in the treatment of hyperthyroidism rats by regulating proliferation, apoptosis, and autophagy of thyroid cell through the mammalian sterile 20-like kinase 1 (MST1)/Hippo pathway.@*METHODS@#Twenty-four rats were randomly divided into 4 groups according to a random number table: control, model group, RS, and RS+Hippo inhibitor (XMU-MP-1) groups (n=6 per group). Rats were gavaged with levothyroxine sodium tablet suspension (LST, 8 μ g/kg) for 21 days except for the control group. Afterwards, rats in the RS group were gavaged with RS extracts at the dose of 1,350 mg/kg, and rats in the RS+XMU-MP-1 group were gavaged with 1,350 mg/kg RS extracts and 1 mg/kg XMU-MP-1. After 15 days of administration, thyroid gland was taken for gross observation, and histopathological changes were observed by hematoxylin-eosin staining. The structure of Golgi secretory vesicles in thyroid tissues was observed by transmission electron microscopy. The expression of thyrotropin receptor (TSH-R) was observed by immunohistochemistry. Terminal-deoxynucleoitidyl transferase mediated nick end labeling assay was used to detect cell apoptosis in thyroid tissues. Real-time quantity primer chain reaction and Western blot were used to detect the expressions of MST1, p-large tumor suppressor gene 1 (LATS1), p-Yes1 associated transcriptional regulator (YAP), proliferating cell nuclear antigen (PCNA), G1/S-specific cyclin-D1 (Cyclin D1), B-cell lymphoma-2 (Bcl-2), Caspase-3, microtubule-associated proeins light chain 3 II/I (LC3-II/I), and recombinant human autophagy related 5 (ATG5). Thyroxine (T4) level was detected by enzyme-linked immunosorbent assay.@*RESULTS@#The thyroid volume of rats in the model group was significantly increased compared to the normal control group (P<0.01), and pathological changes such as uneven size of follicular epithelial cells, disorderly arrangement, and irregular morphology occurred. The secretion of small vesicles by Golgi apparatus was reduced, and the expressions of receptor protein TSH-R and T4 were significantly increased (P<0.01), while the expressions of MST1, p-LATS1, p-YAP, Caspase-3, LC3-II/I, and ATG5 were significantly decreased (P<0.01). The expressions of Bcl-2, PCNA, and cyclin D1 were significantly increased (P<0.01). Compared with the model group, RS extracts reduced the volume of thyroid gland, improved pathological condition of the thyroid gland, promoted secretion of the secretory vesicles with double-layer membrane structure in thyroid Golgi, significantly inhibited the expression of TSH-R and T4 levels (P<0.01), upregulated MST1, p-LATS1, p-YAP, Caspase-3, LC3-II/I, and ATG5 expressions (P<0.01), and downregulated Bcl-2, PCNA, and Cyclin D1 expressions (P<0.01). XMU-MP-1 inhibited the intervention effects of RS extracts (P<0.01).@*CONCLUSION@#RS extracts could inhibit proliferation and promote apoptosis and autophagy in thyroid tissues through MST1/Hippo pathway for treating hyperthyroidism.

Protective effect of iridoid glycosides of radix scrophulariae on endoplasmic reticulum stress induced by oxygen-glucose deprivation and reperfusion / 浙江大学学报·医学版

OBJECTIVE@#To investigate the regulatory effect of iridoid glycoside of radix scrophulariae (IGRS) on endoplasmic reticulum stress induced by oxygen-glucose deprivation and reperfusion @*METHODS@#Rat pheochromocytoma PC12 cells were pretreated with IGRS (50, 100, 200 μg/mL) for 24h, and the @*RESULTS@#The damage caused by OGD/R to PC12 cells was significantly reduced by IGRS, with significant effect on increasing survival rate and reducing LDH release (all @*CONCLUSIONS@#IGRS has neuroprotective effect, which may alleviate cerebral ischemia-reperfusion injury by regulating SERCA2, maintaining calcium balance, and inhibiting endoplasmic reticulum stress-mediated apoptosis.

Study on Radix Scrophulariae for the treatment of hyperthyroidism rat model of yin deficiency based on urine metabonomics / 药学学报

The Chinese herbal Radix Scrophulariae is the main medicine for nourishing yin and reducing fire. It can be used to treat hyperthyroidism due to yin deficiency and fire hyperactivity, but its mechanism is not clear. The present study was aimed to explore the mechanism of Radix Scrophulariae treatment of hyperthyroidism due to yin deficiency and fire hyperactivity. The urine metabolomic approach was conducted using the method of UPLC-TOF-MS. The results showed that Radix Scrophulariae has good therapeutic effects on hyperthyroidism rat model of yin deficiency. After treatment with Radix Scrophulariae, through metabolic profiling and protocol analysis, 6 potential metabolic markers may be closely related with the treatment mechanism of Radix Scrophulariae on this disease, including proline betaine, estrone, thymidine, 5-hydroxyindoleacetic acid, cyclic AMP and L-dopa. The strongest metabolic pathways were associated with tryptophan metabolism, pyrimidine metabolism, tyrosine metabolism, purine metabolism and steroid hormone biosynthesis. The urine metabolomic approach can be applied to clarify the therapeutic mechanism of Radix Scrophulariae on hyperthyroidism rat of yin deficiency, and provide the theoretical basis for the clinical practice of Radix Scrophulariae on nourishing yin to reduce pathogenic fire.

Comparison of two new on-line methods on screening peroxynitrite scavengers in Radix Scrophulariae / 中国药科大学学报

@#Two simple and efficient methods have been developed for screening and identification of natural peroxynitrite scavengers in Radix Scrophulariae (RS). Method I was based on HPLC-DAD-(luminol-peroxynitrite)-CL techniques combined with Q-TOF MS/MS analysis, while method II was based on the pre-column reaction with peroxynitrite followed by HPLC separation with Q-TOF MS/MS analysis. Five active constituents, P1(decaffeoylacteoside), P9(eoside), I6(6″-O-feruloylharpagide), P11(cis-acteoside)and P13(angoroside), were found to possess potential peroxynitrite-scavenging activity by method I, while P9 and P13 were also screened by method II. Method I requires more complex apparatus, but has advantages on simple detection and high sensitivity. Method II requires simpler apparatus than method I, but with more tedious detection and lower sensitivity. However, the methods established above would provide new ways for rapid detection of natural peroxynitrite-scavenging compounds in RX complex matrices.

Simultaneous TLC Identification of Paeoniflorin and Radix Scrophulariae in Yangyin Qingfei Granules / 中国药师

Objective:To improve the TLC identification method in the quality standard for Yangyin Qingfei granules by identifying paeoniflorin and radix scrophulariae on the same condition plate. Methods:The preparation method for paeoniflorin sample solution in the original standard of TLC identification was improved, the extraction method was water-saturated n-butanol extraction, and the reference crude herb solution of radix scrophulariae was prepared. Simultaneous TLC identification of paeoniflorin and radix scrophular-iae was carried out according to the method described in the original standard. Results:The improved TLC method could be used in the simultaneous identification of paeoniflorin and radix scrophulariae, the spots were clear, and the separation and reproducibility were promising without interference. Conclusion:The improved method is more useful in the quality control of the product with simplified operation.

Structural characterization and identification of major constituents in radix scrophulariae by HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry / 中国天然药物

AIM@#To analyze the major constituents in Radix Scrophulariae (Scrophularia ningpoensis).@*METHOD@#Radix Scrophulariae was analyzed by HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (ESI-Q-TOF MS/MS). Compounds were separated by HPLC using a C18 column and gradient elution of acetonitrile and 0.1 % (V/V) acetic acid-water. Negative ion mode was employed.@*RESULTS@#A total of thirty-six compounds, including fourteen iridoid glycosides, nineteen phenylpropanoid glycosides, and three organic acids, were identified from Radix Scrophulariae based on the accurate mass measurement of precursor and product ions. Twenty-one of the constituents were identified by comparing their retention times (tR) and ESI-MS/MS data with those of reference standards and/or previous publications, while another fifteen compounds were tentatively identified or deduced according to their Q-TOF MS/MS data which afforded sufficient structural information.@*CONCLUSION@#It is believed that this study is useful for the identification of constituents in Radix Scrophulariae, as well as related plants and complex prescriptions.

Study on the Chromatography Fingerprint of Radix Scrophulariae by RP-HPLC / 中药新药与临床药理

Objective To establish the chromatography fingerprint of Radix Scrophulariae by RP-HPLC.Methods HPLC was applied in this study.Kromasil KR100-5C18(4.6 mm? 250 mm,5 ? m)column and DAD detector were used with a mixture of methanol and 0.1 % methanoic acid as mobile phase in a gradient mode.Results The chemical substances of Radix Scrophulariae were optimally separated.Conclusion This method is simple,accurate with good reproducibility,thereby can be used specifically for the quality control of Radix Scrophulariae.