Hevea brasiliensis is the main source of natural rubber. Restricted by its tropical climate conditions, the planting area in China is limited, resulted in a low self-sufficiency. Periploca sepium which can produce natural rubber is a potential substitute plant. cis-prenyltransferase (CPT), small rubber particle protein (SRPP) and rubber elongation factor (REF) are key enzymes involved in the biosynthesis of cis-1, 4-polyisoprene, the main component of natural rubber. In this study, we cloned the promoter sequences of CPT, SRPP and REF through chromosome walking strategy. The spatial expression patterns of the three promoters were analyzed using GUS (β-glucuronidase) as a reporter gene driven by the promoters through Agrobacterium-mediated genetic transformation. The results showed that GUS driven by CPT, SRPP or REF promoter was expressed in leaves and stems, especially in the leaf vein and vascular bundle. The GUS activity in stems was higher than that in leaf. This study provided a basis for analyzing the biosynthesis mechanism of natural rubber and breeding new varieties of high yield natural rubber.
Peptide Elongation Factors/genetics* ; Plant Proteins/metabolism* ; Periploca/metabolism* ; Rubber ; Plant Breeding ; Cloning, Molecular

In this study, an ultra-performance liquid chromatography-quadrupole time-of-flight high resolution mass spectrometer(UPLC-Q-TOF-HRMS) was used to investigate the effects of the active ingredients in Periploca forrestii compound on spleen metabolism in rats with collagen-induced arthritis(CIA), and its potential anti-inflammatory mechanism was analyzed by network pharmacology. After the model of CIA was successfully established, the spleen tissues of rats were taken 28 days after administration. UPLC-Q-TOF-HRMS chromatograms were collected and analyzed by principal component analysis(PCA), orthogonal partial least squares discriminant analysis(OPLS-DA), and MetPA. The results showed that as compared with the blank control group, 22 biomarkers in the spleen tissues such as inosine, citicoline, hypoxanthine, and taurine in the model group increased, while 9 biomarkers such as CDP-ethanolamine and phosphorylcholine decreased. As compared with the model group, 21 biomarkers such as inosine, citicoline, CDP-ethanolamine, and phosphorylcholine were reregulated by the active ingredients in P. forrestii. Seventeen metabolic pathways were significantly enriched, including purine metabolism, taurine and hypotaurine metabolism, glycerophospholipid metabolism, and cysteine and methionine metabolism. Network pharmacology analysis found that purine metabolism, glycerophospholipid metabolism, and cysteine and methionine metabolism played important roles in the pathological process of rheumatoid arthritis. This study suggests that active ingredients in P. forrestii compound can delay the occurrence and development of inflammatory reaction by improving the spleen metabolic disorder of rats with CIA. The P. forrestii compound has multi-target and multi-pathway anti-inflammatory mechanism. This study is expected to provide a new explanation for the mechanism of active ingredients in P. forrestii compound against rheumatoid arthritis.
Rats ; Animals ; Periploca ; Cysteine ; Cytidine Diphosphate Choline ; Network Pharmacology ; Phosphorylcholine ; Metabolomics ; Arthritis, Rheumatoid/drug therapy* ; Biomarkers ; Glycerophospholipids ; Methionine ; Purines ; Chromatography, High Pressure Liquid

BACKGROUND: Periploca aphylla is used by local population and indigenous medicine practitioners as stomachic, tonic, antitumor, antiulcer, and for treatment of inflammatory disorders. The aim of this study was to evaluate antidiabetic effect of the extract of P. aphylla and to investigate antioxidant and hypolipidemic activity in streptozotocin (STZ)-induced diabetic rats. METHODS: The present research was conducted to evaluate the antihyperglycemic potential of methanol extract of P. aphylla (PAM) and subfractions n-hexane (PAH), chloroform (PAC), ethyl acetate (PAE), n-butanol (PAB), and aqueous (PAA) in glucose-overloaded hyperglycemic Sprague-Dawley rats. Based on the efficacy, PAB (200 mg/kg and 400 mg/kg) was tested for its antidiabetic activity in STZ-induced diabetic rats. Diabetes was induced via intraperitoneal injection of STZ (55 mg/kg) in rat. Blood glucose values were taken weekly. HPLC-DAD analysis of PAB was carried out for the presence of various polyphenols. RESULTS: HPLC-DAD analysis of PAB recorded the presence of rutin, catechin, caffeic acid, and myricetin. Oral administration of PAB at doses of 200 and 400 mg/kg for 21 days significantly restored (P < 0.01) body weight (%) and relative liver and relative kidney weight of diabetic rats. Diabetic control rats showed significant elevation (P < 0.01) of AST, ALT, ALP, LDH, total cholesterol, triglycerides, LDL, creatinine, total bilirubin, and BUN while reduced (P < 0.01) level of glucose, total protein, albumin, insulin, and HDL in serum. Count of blood cells and hematological parameters were altered in diabetic rats. Further, glutathione peroxidase, catalase, superoxide dismutase, glutathione reductase, and total soluble protein concentration decreased while concentration of thiobarbituric acid reactive substances and percent DNA damages increased (P < 0.01) in liver and renal tissues of diabetic rats. Histopathological damage scores increased in liver and kidney tissues of diabetic rats. Intake of PAB (400 mg/kg) resulted in significant improvement (P < 0.01) of above parameters, and results were comparable to that of standard drug glibenclamide. CONCLUSION The result suggests the antihyperglycemic, antioxidant, and anti-inflammatory activities of PAB treatment in STZ-compelled diabetic rat. PAB might be used as new therapeutic agent in diabetic patients to manage diabetes and decrease the complications.
1-Butanol/chemistry* ; Administration, Oral ; Animals ; Diabetes Mellitus, Experimental/drug therapy* ; Dose-Response Relationship, Drug ; Hypoglycemic Agents/chemistry* ; Male ; Periploca/chemistry* ; Phytochemicals/chemistry* ; Plant Extracts/chemistry* ; Rats ; Rats, Sprague-Dawley ; Streptozocin/adverse effects*

To study the chemical constituents of Periplocae Cortex, the separation and purification of 70% alcohol extract were carried out by column chromatographies on AB-8 macroporous resin, silica gel and preparative HPLC. The structure of the compounds were identified by NMR and TOF-MS. A new compound was isolated and identified as 21-O-methyl-Δ5-pregnene-3β, 14β, 17β, 21-tetraol-20-one-3-O-β-D-oleandropyranosyl(1-->4)-β-D-cymaropyranosyl-(1-->4)-β-D-cymaropyranosyl (1), named as periplocoside P.
Glycosides ; chemistry ; isolation & purification ; Periploca ; chemistry ; Pregnenes ; chemistry ; isolation & purification ; Saponins ; chemistry ; isolation & purification

Ultra performance liquid chromatography (UPLC) was employed for simultaneous determination of three components and fingerprint analysis of Periplocae Cortex with gradient elution of mehtanol and water containing 0.1% phosphoric acid as mobile phase. Three components including chlorogenic acid, 4-methoxysalicylaldehyde and periplocoside were well separated under the analytical condition. Seventeen peaks were selected as the common peaks of 30 batches of Periplocae Cortex. The results showed that there is a significant difference in contents of periplocoside between the samples collected from Henan and Shanxi province. Based on the results of three components quantification and fingerprint analysis, hierarchical clustering analysis ( HCA) and principle component analysis (PCA) were used to further prove the differences between two group samples, and the results indicated that quality of Periplocae Cortex from Shanxi was more stable than that from Henan. The established UPLC fingerprint and quantitative analysis methods could be used efficiently in the quality control of Periplocae Cortex, and this study might contribute to the reasonable clinical application.
Benzaldehydes ; analysis ; China ; Chlorogenic Acid ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Ecosystem ; Periploca ; chemistry ; classification ; growth & development ; Plant Roots ; chemistry ; Quality Control

The chemical constituents of Periploca forrestii were studied by means of macroporous resin, silica gel, ODS column chromatography and PHPLC. Nine compounds were isolated from this plant. By using ESI-MS and NMR, the structures of the nine compounds were determined as scopoletin (1), trans-3, 4-methylenedioxycinnamyl alcohol (2), syringaresinol (3), syringaresinol 4-0-beta-D-glucopyranoside (4), 6'-0-(E)-feruloylsucrose (5), loliolide (6), 4-hydroxy-3-methoxyl-benzaidehyde (7), delta5-pregnene-3beta, 17alpha, 20alpha-triol (8) and delta5-pregnene-3 beta, 17alpha, 20 alpha-triol-20-0-beta-D-canaropyranoside (9), respectively. Compounds 1, 2 and 5-7 are isolated from Periplocagenus for the first time.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Glucosides ; chemistry ; Mass Spectrometry ; Molecular Structure ; Periploca ; chemistry ; Triterpenes ; chemistry

OBJECTIVETo study the chemical constituents of Periploca calophylla.

METHODVarious chromatographic techniques were used to isolate the constituents, and their structures were identified by spectral and chemical methods.

RESULTTwo oligosaccharides were isolated from the chloroform part of P. calophylla and their structures were identified as 4-O-acetyl-beta-cymaropyranosyl (1-->4)-O-beta-D-cymaropyranosyl(1-->4)-O-beta-D-canaropyranosyl (1-->4)-O-beta-D-cymaropyranosy(1-->4)-O-oleandronic acid-delta-lactone(1), and perisaccharide B (2).

CONCLUSIONCompound 1 is a new compound. Compound 2 is reported for the first time from this plant.

Carbohydrate Sequence ; Magnetic Resonance Spectroscopy ; methods ; Molecular Sequence Data ; Molecular Structure ; Oligosaccharides ; analysis ; isolation & purification ; Periploca ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods ; Spectroscopy, Fourier Transform Infrared ; methods

OBJECTIVES: The root barks of Periploca sepium Bge. (P. sepium) has been used in traditional Chinese medicine for healing wounds and treating rheumatoid arthritis. However, toxicity in high-doses was often diagnosed by the presence of many glycosides. The potential mutagenicity of P. sepium was investigated both in vitro and in vivo. METHODS: This was examined by the bacterial reverse mutation (Ames) test using Escherichia coli WP2uvrA and Salmonella typhimurium strains, such as TA98, TA100, TA1535, and TA1537. Chromosomal aberrations were investigated using Chinese hamster lung cells, and the micronucleus test using mice. RESULTS: P. sepium did not induce mutagenicity in the bacterial test or chromosomal aberrations in Chinese hamster lung cells, although metabolic activation and micronucleated polychromatic erythrocytes were seen in the mice bone marrow cells. CONCLUSIONS: Considering these results, it is suggested that P. sepium does not have mutagenic potential under the conditions examined in each study.
Animals ; Arthritis, Rheumatoid ; Biotransformation ; Bone Marrow ; Chromosome Aberrations ; Cricetinae ; Cricetulus ; Erythrocytes ; Escherichia coli ; Glycosides ; Lung ; Medicine, Chinese Traditional ; Mice ; Micronucleus Tests ; Periploca ; Salmonella typhimurium

OBJECTIVETo study the chemical constituents of Periploca forrestii.

METHODThe constituents were separate using such various column chromatographic techniques as silica gel, RP-18 silica gel, MCI and Sephadex LH-20. Their structures were identified by such methods as spectral analysis.

RESULTTen compounds were isolated and identified as periforgenin A-3-O-beta-digitoxopyranoside (1), beta-sitosterol (2), periforoside I (3), ursolic acid (4), periplogenin (5), periplocin (6), glycoside E (7), periplocoside M (8) , daucosterol (9), 2alpha, 3alpha, 23-trihydroxy-urs-12-en-28-oic acid (10).

CONCLUSIONCompound 1 was a new cardiac glycoside and compound 8 was reported for the first time from this plant.

Cardiotonic Agents ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Molecular Structure ; Periploca ; chemistry

OBJECTIVETo study the characteristics of the growth and periplocin accumulation of the adventitious roots of Periploca sepium, and on this basis, study the effect of Ag+ and La3+ elicitors on the growth and periplocin accumulation of the adventitious roots.

METHODThe adventitious roots were sampled every four days, and the dry weight and the contents of the periplocin were measured. The curves of the growth and periplocin accumulation of the roots were plotted. The Ag+ and La3+ elicitors with different concentrations were added to the medium when the adventitious roots grew in the stable phase to study the optimal concentration which was good to synthesize the periplocin. Besides, the optimal concentration of Ag+ and La3+ elicitors was added to the different growth phases to study the effect of the elicitors on the growth and periplocin synthesis of adventitious roots.

RESULTThe characteristics of the growth of adventitious roots of P. sepium showed a typical growth S-Curve, which displayed a half-coupling relationship with the metabolism of periplocin. The optimal concentrations of Ag+ and La3+ elicitors were both 0.05 mmol L(-1). Besides, it was the best period for the Ag+ and La3+ elicitors to elicit the synthesis of periplocin when in the terminally exponential phase.

CONCLUSIONThe growth of adventitious roots and the accumulation of periplocin show a half-coupling relationship. Besides, the concentration and additive time of Ag and La3+ elicitors obviously influences the growth of adventitious roots and synthesis of periplocin.

Drugs, Chinese Herbal ; analysis ; metabolism ; Lanthanum ; metabolism ; Periploca ; chemistry ; growth & development ; metabolism ; Plant Roots ; chemistry ; growth & development ; metabolism ; Saponins ; analysis ; metabolism ; Silver ; metabolism