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Disruption of the ubiquitin-mediated proteolysis pathway: a study of seed aging in Saposhnikovia divaricata caused by UBC1 gene family suppression.

BACKGROUND: Saposhnikovia divaricata (Turcz.) Schischk. is a perennial herb whose seed aging during storage significantly reduces germination rates, limiting industrial-scale production. Reactive oxygen species (ROS)-induced oxidative damage is a key driver of seed aging, but the underlying mechanisms in Saposhnikovia divaricata remain unclear. RESULTS: Suppression of the UBC1 gene family reduces the activity of ubiquitin-conjugating enzymes, leading to dysfunction of the ubiquitin-mediated proteolysis pathway, which in turn decreases protein degradation efficiency and causes the accumulation of damaged proteins. Transcriptome analysis revealed predominant downregulation of genes crucial for seed physiological maintenance. By the fourth year of storage, germination dropped sharply to 30.67%, accompanied by embryo cavitation. Downregulation of ribosome pathway genes hindered ribosome assembly and protein synthesis, while suppression of endoplasmic reticulum protein processing genes led to unfolded/misfolded protein accumulation and intensified cellular stress, accelerating aging. Proteomic analysis showed increased total differential and antioxidant-related proteins. ROS content fluctuated with storage time: peroxyl radicals peaked in year two (5.68 RFU/mg), whereas hydroxyl radicals and hydrogen peroxide were highest in year four (0.0655 pg/mL and 0.0946 pg/mL, respectively), with significant differences across periods. Elevated membrane-related proteins, increased electrical conductivity, and malondialdehyde content (maximum 54.30 nmol/g at year four) confirmed oxidative membrane damage. ROS-induced stress promotes protein misfolding, and reduced UBC1 expression is associated with impaired clearance of misfolded proteins by the ubiquitin-mediated proteolysis pathway. CONCLUSIONS: This study provides the first integrated transcriptomic and proteomic insight into UBC1 deficiency-mediated seed aging in Saposhnikovia divaricata. The findings enhance molecular understanding of seed aging and offer new directions for improving seed storage and viability.

Ubiquitin-Conjugating Enzymes↗

[Studies on the method for the processing roots of cultivated Saposhnikovia divaricata].

OBJECTIVE: To establishing a standard processing method for cultivated Saposhnikovia divaricata. METHOD: Crude drug was dried at different temperature, and the contents of prim-O-glucosyl-cimifugin,cimifugin,4'-O-beta-glucosyl-5-O-methylvisamminol, sec-O-glucosyl-hamaudol in the processed crude drug were determined by HPLC. RESULT: Of the contents of the four compounds, the sample that heated by 45 degrees C was at the best level. Prim-O-glucosggl-cinmifugin was 0.636%, cimifugin was 0.043%, 5-O-methtlvsamminol was 0.598%, sec-O-glu-cosyl-hamaudol was 1.85%, and total content was 1.302%. CONCLUSION: The best drying temperature for processing is 45 degrees C.

Apiaceae↗

An acidic polysaccharide having activity on the reticuloendothelial system from the roots and rhizomes of Saposhnikovia divaricata.

An acidic polysaccharide, named saposhnikovan C, was isolated from the roots and rhizomes of Saposhnikovia divaricata Schischk. It was homogeneous as judged by electrophoresis and gel chromatography, and showed remarkable reticuloendothelial system-potentiating activity in a carbon clearance test. It is composed of D-galacturonic acid:L-rhamnose:L-arabinose:D-galactose in a molar ratio of 27:7:8:8, and its molecular mass was estimated to be 132000. About 30% of the D-galacturonic acid residues exist as the methyl esters. O-Acetyl groups were identified, and the content amounted to 3.3%. Methylation analysis, carbon-13 nuclear magnetic resonance, and controlled Smith degradation studies indicated the structural features. It has a pectin-like rhamnogalacturonan backbone with branched arabinan and galactan side chains.

Carbohydrate Sequence↗

The major pectic arabinogalactan having activity on the reticuloendothelial system from the roots and rhizomes of Saposhnikovia divaricata.

The major acidic polysaccharide, named saposhnikovan A, was isolated from the roots and rhizomes of Saposhnikovia divaricata. It showed remarkable reticuloendothelial system-potentiating activity in a carbon clearance test. It is composed of L-arabinose: D-galactose: D-galacturonic acid in the molar ratio of 6:15:10, and its molecular weight was estimated to be 54000. About 35% of the D-galacturonic acid residues exist as the methyl esters. Methylation analysis, 13C-nuclear magnetic resonance, and controlled Smith degradation studies indicated that the polysaccharide has the alpha-1----4-linked D-galacturonan backbone bearing alpha-1----5-linked L-arabino-beta-3,6-branched D-galactan side chains.

Animals↗

Analgesic components of saposhnikovia root (Saposhnikovia divaricata).

By activity-oriented separation using the writhing method in mice, the analgesic components of Saposhnikovia root (Saposhnikovia divaricata Schischkin; Umbelliferae) were identified to be chromones, coumarins, polyacetylenes and 1-acylglycerols. Two new components, divaricatol and (3'S)-hydroxydeltoin, were also isolated. The most potent analgesia was observed in chromones such as divaricatol, ledebouriellol and hamaudol, which inhibited writhing inhibition at an oral dose of 1 mg/kg in mice. Acylglycerols also showed inhibition significantly at a dose of 5 mg/kg. In some pharmacological tests using sec-O-glucosylhamaudol, the compound showed analgesia by the tail pressure and the Randall & Selitto methods, and its writhing inhibition was not reversed by naloxone.

Analgesics, Non-Narcotic↗

Inducible nitric oxide synthase inhibitors from Saposhnikovia divaricata and Panax quinquefolium.

A series of polyacetylenes, falcarinone, panaxynol, falcarindiol, panaxydol, and panaxytriol, were isolated from Saposhnikovia divaricata (Turcz.) Schischk and Panax quinquefolium L. These polyacetylenes were identified as active principles on the inhibition of nitrite production by inducible nitric oxide synthase (iNOS). Treatment with 10 microM of panaxynol, falcarindiol, panaxydol and panaxytriol decreased the LPS/IFN-gamma-stimulated accumulation of nitrite by 71.92 +/- 3.07, 69.95 +/- 3.68, 45.48 +/- 6.11 and 36.85 +/- 8.80%, respectively. The IC50 value of falcarinone, panaxynol, falcarindiol, panaxydol and panaxytriol was > 20, 2.23, 1.98, 6.58 and 9.85 microM, respectively.

Apiaceae↗

[Determination of four components in root of Saposhnikovia divaricata by HPLC gradient elution].

OBJECTIVE: To develop an HPLC method for determination of components in root of Saposhnikovia divaricata. METHOD: The separation of four components was achieved on a reversed-phase Alltima C18 column with MeOH-H2O gradient elution at a flow rate of 1.0 mL x min(-1). RESULT: Six samples were determined and there is no distinct difference between the contents determined by the new and old methods. CONCLUSION: The new method is more simple and convenient than the old method.

Apiaceae↗

[Comparative studies on the febrifugal analgesic and anticonvulsive activities of water extracts from cultivated and wild Saposhnikovia divaricata].

In this paper, comparative studies on main pharmacological activities and toxilogical activities of water extracts from cultivated Saposhnikovia divaricata (SD) and wild SD in Qixia county of Shandong province were performed. The results showed that the febrifugal analgesic and anticonvulsived activities of water extracts from cultivated SD and wild SD were primary same. Estimative results of LD50 showed that order of toxicity were such as firstly direct cultivated SD, secondly cuttage SD and finally wild SD. Therefore it was recognized that cultivated SD completely can replaced wild one in medical use.

Animals↗

[GC-MS analysis of essential oils from the roots of Saposhnikovia divaricata (Turcz.) Schischk, Libanotis laticalycina Shan et Sheh, Seseli yunnanense Franch. and Peucedanum dielsianum Fedde ex Wolff].

OBJECTIVE: To analyze the constituents of the essential oils from the roots of Saposhnikovia divaricata, Libanotis laticalycina, Seseli yunnanense and Peucedanum dielsianum. METHOD: GC-MS method was used. RESULT: 58 compounds were identified. CONCLUSION: To compare the constituents and their contents of 6 samples, 3 samples of S. divaricata growing in 3 places are similar, L. laticalycina, S. yunnanense and P. dielsianum are different from S. divaricata in varying degrees.

Alkynes↗

Inducible nitric oxide synthase inhibitor of the Chinese herb I. Saposhnikovia divaricata (Turcz.) Schischk.

L-Arginine derived nitric oxide (NO) and its derivatives, such as nitrogen dioxide and peroxynitrite, play a role in inflammation and also possibly in the multistage process of carcinogenesis. Four furanocoumarins and eight chromones isolated from the dried root of Saposhnikovia divaricata (Fang Feng in Chinese) and evaluated for their effects on the synthesis of NO induced by lipopolysaccharide (LPS) in macrophage cell line RAW 264.7. The inhibition of nitrite production, as an index for NO released from the macrophage cells, was quantitatively analyzed by Griess reaction. The results showed that imperatorin and deltoin are potential NO production inhibitor, and their IC50 values for inhibition of nitrite production were 17.3 and 11.6 microg/ml, respectively. Western-blot analysis demonstrated that iNOS enzyme activity was not inhibited by treatment with imperatorin or deltoin, but revealed that both compounds inhibited the expression of the iNOS protein.

Animals↗

[Studies on quality specification standards for Saposhnicovia divaricata (Turcz.) Schischk].

OBJECTIVE: To work out a qualitative-quantitative standard for Saposhnikovia divaricata. METHOD: TLC and HPLC were adopted. RESULT: Four chromone compounds were identified by TLC, and the contents of two chromone compounds were determined by HPLC. CONCLUSION: Prim-O-glucosylcimifugin, 4'-O-beta-D-glucosyl-5-O-methylvisamminol, sec-O-glucosylhamaudol and cimifugin should be present simultaneously in S. divaricata. The contents of prim-O-glucosylcimifugin and 4'-O-beta-D-glucosyl-5-O-methylvisamminol in S. divaricata should not be lower than 0.12% respectively.

Apiaceae↗

Inhibitory potential of herbal medicines on human cytochrome P450-mediated oxidation: properties of umbelliferous or citrus crude drugs and their relative prescriptions.

To investigate the possible drug interaction with herbal medicine, hot water decoctions or 40% ethanol infusions of several Umbelliferous or Citrus crude drugs and their prescriptions were examined in vitro for their abilities to inhibit human cytochrome P450 3A (CYP3A). Addition of each decoction or infusion from Baizhi (Angelica dahurica and varieties), Qianghuo (Notopterygium incisum or N. forbesii), Duhuo (Angelica biserrata), Fangfeng (Saposhnikovia divaricata), Danggui (Angelicasinensis), Zhishi or Zhiqiao (Citrus aurantium) resulted in various degrees of human CYP3A inhibition as determined by microsomal testosterone 6beta-hydroxylation. The inhibitory potency was consistent with the abundance of the hydrophobic components for each sample. Experiments on the infusion of a Japanese Baizhi (BZ1) showed the major role of furanocoumarins on human CYP3A inhibition. Some of the crude drugs and a related prescription showed increased inhibition after the preincubation, suggesting the involvement of a mechanism-based inhibition. Some formulated prescriptions, however, showed intense inhibition with their hydrophobic fractions rather than with their hydrophobic fractions, suggesting that components other than furanocoumarins in herbal prescriptions may also cause CYP3A inhibition. These results indicate the necessity of intensive investigations on the possible drug interaction with traditional medicines.

Aryl Hydrocarbon Hydroxylases↗

[Herbalogic studies on the Chinese drug fangfeng].

Through a critical study of the descriptions of Fangfeng in the ancient Chinese herbals, it may be concluded that the chief botanical source of this drug was Saposhnikovia divaricata, besides, the roots of Peucedanum ledebourielloides and P. wawrii may also be used. The root of Glehnia littoralis may be used as an substitute and called Shifangfeng. The difference in geographical sources of Fangfeng in ancient and present times is also discussed.

China↗