Catalpol as a Component of Rehmannia glutinosa Protects Spinal Cord Injury by Inhibiting Endoplasmic Reticulum Stress-Mediated Neuronal Apoptosis.

Disturbance of the internal environment in the spinal cord after spinal cord injury (SCI) is an important cause of the massive death of neurons in the injury area and one of the major problems that lead to the difficult recovery of motor function in patients. Rehmannia glutinosa, a famous traditional Chinese medicine, is commonly used in neurodegenerative diseases, whereas an iridoid glycoside extract of catalpol (CAT), with antioxidant, antiapoptotic, and neuroprotective pharmacological effects. However, the neuroprotective and anti-apoptosis mechanism of CAT in SCI remains unclear. In our study, we found that CAT has a restorative effect on the lower limb motor function of rats with SCI by establishing a rat model of SCI and treating CAT gavage for 30 days. Our study further found that CAT has the effect of inhibiting apoptosis and protecting neurons, and the action pathway may reduce endoplasmic reticulum (ER) stress by inhibiting CHOP and GRP78 expression and then reduce apoptosis and protect neurons through the Caspase3/Bax/Bcl-2 pathway. In conclusion, we demonstrated that CAT can treat SCI by inhibiting ER stress-mediated neuronal apoptosis and has the potential to be a clinical drug for the treatment of SCI.

[Comparative study on HPLC fingerprints between crude and processed Ligustri Lucidi Fructus].

To establish high performance liquid chromatography(HPLC) fingerprints for crude and processed Ligustri Lucidi Fructus,and to evaluate their quality through the similarity calculation and chemical pattern recognition. The separation was performed with Syncronis C_(18) column(4.6 mm × 250 mm, 5 µm), with acetonitrile(A) and 0.1% phosphoric acid solution(B) as the mobile phase for gradient elution, and a detection wavelength of 280 nm. HPLC was used to detect 22 batches of crude and processed Ligustri Lucidi Fructus,and the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(2012 Edition) was used to evaluate the similarity among 22 batches. The research on pattern recognition was conducted with cluster analysis(CA), principal component analysis(PCA), and partial least squares discriminate analysis(PLS-DA). HPLC fingerprints of crude and processed Ligustri Lucidi Fructus were established, with similarity ranging from 0.9 to 1.0. The crude and processed Ligustri Lucidi Fructus can be obviously distinguished by using CA, PCA and PLS-DA. According to the results of PLS-DA,11 constituents including hydroxytyrosol, tyrosol, specnuezhenide and oleuropein were the main marker components leading to the difference. The established fingerprint method is stable and reliable, and can provide method basis for quality control of crude and processed Ligustri Lucidi Fructus. Chemical pattern recognition is proved to be helpful in comprehensive quality control and evaluation of Ligustri Lucidi Fructus before and after the process.

Mechanism-based anti-anxiety effects of polysaccharides extracted from shudihuang (radix rehmanniae preparata) by two-dimensional electrophoresis analysis in rat hippocampus proteins.

OBJECTIVE: To investigate mechanism-based anti-anxiety effects of Shudihuang (Radix Rehmanniae Preparata) polysaccharides (RRPPs) through two-dimensional electrophoresis (2-DE) analysis with mass spectrometry (MS) of hippocampus proteins in rats treated with monosodium L-glutamate (MSG). METHODS: MSG (4 g/kg) or normal saline (NS) was injected subcutaneously into infant male rats on days 2, 4, 6, 8, 10 after birth. MSG-treated rats at 8 weeks old were given NS, diazepam, or RRPPs daily for seven consecutive days by intragastric administration, while NS-treated rats given the same volume of NS. Elevated plus maze (EPM) and light/dark transition (LDT) tests were used to observe anti-anxiety effects of RRPPs at 1 h after the last administration. After EPM and LDT tests, hippocampus tissues were excised on ice rapidly from the brains of rats. Thereafter, 2-DE and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) were used for detecting differential proteins in hippocampus of rats so as to explore the potential mechanisms. RESULTS: RRPPs (200, 400 mg/kg) could significantly inhibit MSG-induced decrease of time and entries percentages in open zones in EPM test and numbers of light-dark transition in LDT test. Further analysis of 2-DE and MALDI-TOF/MS indicated that beta-synuclein, protein DJ-1, peroxiredoxin-2, peroxiredoxin-6, dimethylarginine dimethylaminohydrolase 1 (DDAH-1) and iron-sulfur proteins were all found to be down-regulated significantly in MSG-treated rats, while such down-regulation was significantly inhibited after treatment with RRPPs. CONCLUSION: RRPPs showed anti-anxiety effects and potential mechanisms might be related to inhibiting MSG-induced down-regulation of beta-synuclein, DJ-1, peroxiredoxin-2, peroxiredoxin-6, DDAH-1 and iron-sulfur proteins in hippocampus of rats.

[Fingerprint research on immunosuppressive fraction of Rehmanniae Radix by HPLC].

OBJECTIVE: To study the HPLC fingerprint of immunosuppressive fraction of Rehmanniae Radix, the genuine medicinal materials in Henan province. METHODS: By HPLC method,Dikma Platisil-C18 column (250 mm x 4.6 mm, 5 microm) was used with acetonitrile-0.1% phosphoric acid as mobile phase by gradient elution. The detection wavelength was 210 nm, flow rate was 1.0 mL/min and column temperature was set at 30 degrees C. RESULTS: Compared the 10 batches sample of the fingerprint similarity, 18 common peaks were marked and five peaks were identified. The similarities were more than 0.9. CONCLUSION: HPLC fingerprint of immunosuppressive fraction of Rehmanniae Radix is established, which lays the foundation for further pharmacodynamics research on Rehmanniae Radix.

[Study on the triterpenoids from the fruits of Ligustrum lucidum].

OBJECTIVE: To study the triterpenoids from the fruits of Ligustrum lucidum. METHODS: The constituents were extracted with 95% EtOH, partitioned with different solvents, isolated and purified by silica gel, Sephadex LH-20 and PHPLC column chromatography. The structures were elucidated by physicochemical properties and spectroscopic data. RESULTS: Thirteen compounds were isolated and identified as methyl oleanolate (1), 3-O-acetylursolic acid (2), 3-ketooleanolic acid (3), 19alpha-hydroxyl ursolic acid (4), lupeol (5), 3-O-acetyl oleanolic acid (6), oleanolic acid (7), betulin (8), ursolic acid (9), fouquierol (10), 19alpha-hydroxyl acetylursolic acid (11), 3-O-cis-P-coumaroyl maslinic acid (12) and 3-O-trans-P-coumaroyl maslinic acid (13). CONCLUSION: Compounds 1 - 4 are obtained from this plant for the first time.

[Experimental research about the effect of Rehmannia glutinosa on the drug effect fingerprints of the rabbit's blood serum of Yin deficiency and heat flourishing syndrome].

OBJECTIVE: To study about the effect of Rehmannia glutinosa Libosch on the drug effect fingerprints of the rabbit's blood serum of yin deficiency and heat flourishing syndrome. METHODS: The infusion of furosemide and escherichia coli endotoxin were used to replicate rabbit' s yin deficiency and heat flourishing syndrome animal model; high performance liquid chromatography gradient elution was adopped to study the change of the rabbit serum drug effect fingerprint in copying between before and after model-making and administration. RESULTS: Rehmannia glutinosa Libosch mainly affected metabolism level of the inner substance with the function of nourishing yin clearing heat. CONCLUSION: The serum HPLC fingerprint established is capable to reflect the function of nourishing yin clearing heat of Rehmannia glutinosa Libosch; the further study on its drug effect fingerprint will help to clarify its activity and metabolism of action.