Cloning and characterization of the promoters of the key genes CPT, SRPP and REF involved in Periploca sepium rubber biosynthesis / 生物工程学报

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.

Mechanism of active ingredients in Periploca forrestii compound against rheumatoid arthritis based on integrative metabolomics and network pharmacology / 中国中药杂志

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.

Phytochemicals of Periploca aphylla Dcne. ameliorated streptozotocin-induced diabetes in rat

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.

A new C21 steroidal saponins from Periplocae Cortex / 中国中药杂志

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.

Quality assessment of Periplocae Cortex from different habitats by UPLC fingerprint and quantitative analysis / 中国中药杂志

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.

Chemical constituents of Periploca forrestii / 中国中药杂志

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.

Lack of Mutagenicity Potential of Periploca sepium Bge. in Bacterial Reverse Mutation (Ames) Test, Chromosomal Aberration and Micronucleus Test in Mice

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.

A new cardiac glycoside from Periploca forrestii / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of Periploca forrestii.</p><p><b>METHOD</b>The 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.</p><p><b>RESULT</b>Ten 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).</p><p><b>CONCLUSION</b>Compound 1 was a new cardiac glycoside and compound 8 was reported for the first time from this plant.</p>

A new oligosaccharide from Periploca calophylla / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of Periploca calophylla.</p><p><b>METHOD</b>Various chromatographic techniques were used to isolate the constituents, and their structures were identified by spectral and chemical methods.</p><p><b>RESULT</b>Two 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).</p><p><b>CONCLUSION</b>Compound 1 is a new compound. Compound 2 is reported for the first time from this plant.</p>

Affect of Ag+ and La3+ elicitors on growth and accumulation of adventitious roots of Periploca sepium / 中国中药杂志

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHOD</b>The 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.</p><p><b>RESULT</b>The 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.</p><p><b>CONCLUSION</b>The 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.</p>

A diterpenoid quinone from Periploca forrestii / 中国中药杂志

Tanshinone II A, which was known unique to the salvia, was separated and purified by silica gel column chromatography and recrystallisation from an ethyl acetate-soluble portion (the anti-inflammatory active portion) of ethanol extract of Periploca forrestii. The diterpenoid quinone was obtained from the Periploca for the first time.

Induction of Periploca sepium in vitro plantlet and studies on dynamic accumulation of periplocin / 中国中药杂志

<p><b>OBJECTIVE</b>To analyze the content of periplocin in different part of the Periploca sepium in vitro plantlet and study its dynamic variation during the process of differentiation.</p><p><b>METHOD</b>The seeds were generated seedling under aseptic condition, and the cut hypocotyl was induced to form the callus and adventitious buds on the MS culture medium with the hormone of IBA 0.1 mg x L(-1) + BA 1 mg x L(-1). The seedling was cut down when the buds grew up to 3 cm and then the root was cultured in the 1/2 MS culture medium with the hormone of IBA 0.5 mg x L(-1) to form intact plantlet. Different parts of it were collected and the content of periplocin was measured during the process of differentiation.</p><p><b>RESULT</b>The contents of periplocin varied widely in different parts during the process of differentiation, with the highest in the roots and then callus, stem and leaf of intact plantlet, stem and leaf of plantlet without root from high to low.</p><p><b>CONCLUSION</b>The periplocin of the secondary metabolite is more likely to be produced and accumulated in root and callus. Periplocin in stem and leaf is probably transported by conducting tissue.</p>

Determination of quercetin-3-O-alpha-L-arabinopyranoside in Periploca forrestii by RP-HPLC / 中国中药杂志

The aim of the paper was to develop a HPLC method for the quality control of Periploca forrestii Schltr. The 18 samples were analyzed on a Hypersil C18 column. The mobile phase was methanol-water (33:67) and the flow rate was 1 mL x min(-1). The detection wavelength was at 370 nm and column temperature was 25 degrees C. The linear relationship was good (r = 0.999 9) in the range of 0.204 4-2.044 microg for quercetin-3-O-alpha-L-arabinopyranoside. The average recovery was 97.78% (RSD 0.8%, n = 9). The contents of 18 samples varied from 0.171% to 0.264%. The method showed high precision, good repeatability and stability, so it can be used to assess the quality of P. forrestii.

Glycosides from Periploca forrestii / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents from n-BuOH fraction of the stems of Periploca forrestii.</p><p><b>METHOD</b>The compounds were separated and purified by various chromatographic techniques and recrystallisation. The physico-chemical properties and spectral data were applied to identify structure of these compounds.</p><p><b>RESULT</b>Six glycosides were isolated and identified as emodin-8-O-beta-D-glucopyranoside (1), physcion-8-O-beta-D-glucopyranoside (2), kaempferol-3-O-beta-D-galactopyranoside (3), kaempferol-3-O-alpha-L-arabinopyranoside (4), quercetin-3-O-beta-D-glucopyranoside (5), quercetin-3-O-alpha-L-arabinopyranoside (6).</p><p><b>CONCLUSION</b>These six compounds were obtained for the first time from this plant.</p>

Chemical constituents of Periploca forrestii and their cytotoxicity activity / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the chemical constituents of the roots of Periploca forrestii and evaluate their cytotoxicity activities.</p><p><b>METHOD</b>Silica gel column chromatography was employed for the isolation and purification of chemical constituents. The structures were identified on the basis of spectral data and the cytotoxicities of compounds 2-4 were investigated by several tumors cell lines including blood tumor (HL-60, CCRT-CEM), prostate tumor (PC-3, DU-145) and Melanoma (UACC-62).</p><p><b>RESULT</b>Four compounds were isolated and identified as follows, lupeol-20(29)-en-3-nonadecanoate (1), peroiforoside I (2), 3beta,5beta,14beta-3OH-8beta-H-car-20(22)-enolide (3), perplocin (4).</p><p><b>CONCLUSION</b>Compound 1 is a new lupane triterpene fatty acid ester. Compounds 2-4 showed notable cytotoxicity against all tumor lines.</p>

Chemical constituents of Periploca forrestii / 中国中药杂志

<p><b>OBJECTIVE</b>To study the constituents of the stems of Periploca forrestii.</p><p><b>METHOD</b>The compounds were separated and purified by silica gel column chromatography, recrystallization and high-performance liquid chromatography. The structures were identified by various spectroscopic methods.</p><p><b>RESULT</b>Nine compounds were isolated and identified as 3-O-acetyloleanolic acid (1), 14-ursen-3-ol-1-one (2), taraxasterol (3), jacoumaric acid (4), periplogenin (5), 2alpha,3beta-dihydroxyursolic acid (6), E-p-hydroxy-cinnamic acid (7), caffeic acid (8), proanthocyanidin A2 (9).</p><p><b>CONCLUSION</b>All compounds except 6 were isolated from this plant for the first time, compound 4, 9 were obtained from the Periploca for the first time.</p>

Studies on content determination of polysaccharides in Periploca forrestii / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a method for content determination of polysaccharides in Periplocaforrestii.</p><p><b>METHOD</b>The conversion coefficient of P. forrestii. polysaccharide to glucose was obtained by refined polysaccharides, and then the content of crude polysaccharides was determined by sulfuric-phenol method.</p><p><b>RESULT</b>The content of crude polysaccharides in P. forrestii. was 54.68% (RSD 3.0%, n = 6), and the average recovery was 101.7% (RSD 6.0%, n =6).</p><p><b>CONCLUSION</b>The method was simple, rapid, and accurate.</p>

Chemical constituents from root barks of Periploca sepium / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the chemical constituents of the root barks of Periploca sepium.</p><p><b>METHOD</b>Column chromatographic techniques were used to isolate the chemical constituents. NMR and MS methods were employed for their structural elucidation.</p><p><b>RESULT</b>Eight compounds were isolated and identified as isovanillin (1), vanillin (2), 4-methoxysalicylic acid (3), (24R)-9, 19-cycloart-25-ene-3beta, 24-diol (4), (24S)-9, 19-cycloart-25-ene-3beta, 24-diol (5), cycloeucalenol (6), beta-amyrin acetate (7) and alpha-amyrin (8).</p><p><b>CONCLUSION</b>Compounds 1-6 were isolated from this plant for the first time.</p>

Clinical efficacy of compound periploca liquid in treating condyloma acuminatum and its effect on human papilloma virus DNA expression in vitro / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the clinical efficacy of compound periploca liquid (CPL) in treating condyloma acuminatum (CA), and to explore its mechanisms at molecular level.</p><p><b>METHODS</b>Eighty-one patients with CA were randomly divided into three groups, 30 patients in Group A were treated with CPL, 21 in Group B were treated with periploca syrup and 30 in Group C were treated with Youtuoxin (YXG). The clinical efficacy and adverse reaction occurred in the three groups was evaluated respectively. Besides, change of human papilloma virus (HPV) DNA in two patients with CA was determined by polymerase chain reaction (PCR) in vitro after being treated with CPL and periploca, the monarchic drug of CPL.</p><p><b>RESULTS</b>The cure rate obtained in group A, B, and C was 56.67%, 42.86% and 63.33% respectively, the total effective rate in them was 83.33%, 71.43% and 86.67% respectively, the difference of therapeutic efficacy was insignificant among the three groups. But the adverse reaction occurrence in them (6.67%, 4.76% and 86.67%) was significantly different (P < 0.01). The recurrent rate in them was 14.29%, 12.5% and 47.1% respectively. PCR showed negative expression of HPV in the two samples of CA of same concentration after the verrucous homogenate suspension being treated with CPL or periploca syrup of different concentration, while it was positive after the suspension was treated with the group of normal saline without any drug.</p><p><b>CONCLUSION</b>The therapeutic efficacies of CPL, periploca syrup and Youtuoxin are not significantly different, but the former two have advantages of less adverse effects and lower recurrent rate. And they are possibly having germicidal action on HPV-DNA in CA tissue in vitro.</p>

Isolation and structure elucidation of glycosides in n-butanol extracts from rhizome of Periploca calophylla / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents from n-butanol extracts of Periploca calophylla.</p><p><b>METHOD</b>The constituents were isolated and purified by chromatographic technology and their structures were elucidated on the basis of physicochemical property and spectroscopic methods.</p><p><b>RESULT</b>Eight glycosides were isolated and identified as periplocin (I), kaempferol 3-alpha-D-arabinoside (II), kaempferol 3-O-beta-D-glucoside (III), 3',4',5,7-tetrahydroxyflavanone-2(S)-3'-O-beta-D-glucopyranoside (IV), (+)-syringaresinol-4'-O-beta-D-monoglucoside (V), 1-sinapoylglucoside (VI), erigeside C (VII), 2,6-dimethoxy-4-hydroxyphenol 1-O-beta-D-glucoside (VIII).</p><p><b>CONCLUSION</b>All the compounds were isolated for the first time from this plant.</p>