To investigate the effect of acid and alkali stress on ginsenoside content of Panax ginseng, adventitious roots culture in bioreactors were incubated for 30 d and pH value was adjusted. Ginsenoside content increased by reducing or raising the pH in culture medium, the muxium ginsenoside content was determined on the 5th days after acid treatment and on the 7th days after alkali treatment. The result of histochemical localization of ginsenoside revealed that the red color from light to dark were found in the adventitious root tissue, and ginsenoside mainly located in the pericycle cells where appeared the dark red color.
Ginsenosides ; metabolism ; Hydrogen-Ion Concentration ; Panax ; metabolism ; physiology ; Plant Roots ; metabolism ; Stress, Physiological ; Time Factors

To improve cell suspension culture system of Panax ginseng, the dynamic of cell growth and medium consumption were studied, and the effects of filter on the culture vessel, revolution number, and inoculation density on cell growth and ginsenoside accumulation were also investigated. The maximum cell growth and ginsenoside accumulation was found on the 20th days of suspension culture, therefore, 20 days were confirmed as a suitable culture period for mass production of ginsenoside. Cell growth and ginsenoside content were promoted when the culture vessel had a ventilated filter. Revolution speed during suspension culture affected cell growth, but not ginsenoside content, a peak of ginsenoside productivity was found in the treatment of 120 r x min(-1). Inoculation density also influenced cell growth and ginsenoside accumulation, inoculation density of 6 g was better than other inoculation densities, the ginsenoside content and productivity were up to 12.8 mg x g(-1) DW and 146.6 mg x L(-1), respectively.
Cell Culture Techniques ; methods ; Cell Proliferation ; Culture Media ; chemistry ; Ginsenosides ; metabolism ; Panax ; cytology ; growth & development ; metabolism ; Suspensions

OBJECTIVETo provide a new material for producing the Rhodiolasachalinensis products, the effect of methyl jasmonate (MeJA) on callus biomass and effective compound accumulation of Rhodiolasachalinensis was studied.

METHODThe calluses-cultured in 3 L-air lift balloon type bioreactor were treated with MeJA after 20 d of bioreactor culture and the effect of MeJA concentration and treatment days on callus biomass, salidroside or polysaccharide accumulation and superoxide dismutase (SOD) and peroxidase (POD) activities were investigated.

RESULTThe callus biomass was not significantly different after MeJA treatment (125) for 0-6 d but obviously decreased after 6 d treatment. The maximum salidroside or polysaccharide contents and SOD or POD activities were found after 4 d treatment of MeJA. MeJA concentration significantly affected callus biomass and effective compound accumulation, biomass decreased at MeJA concentrations higher than 125 μmol x L(-1). However, the effective compound contents were determined at higher MeJA concentration, and the highest salidroside and polysaccharide accumulation was found at 225 and 275 μmol x L(-1) MeJA, respectively and the maximum SOD and POD activities was found at 225 μmol x L(-1) MeJA. The effective compound contents in callus were compared with field-grown plants. Salidroside contents in calluses were 1.1-fold and 2. 4-fold more than in plant roots and stem or leave, respectively. Polysaccharide content in calluses were 3. 6-fold and 8.0-fold more than in plant roots and stem or leave, respectively.

CONCLUSIONSalidorside and polysaccharide in Rhodiolasachalinensiscalluses improved by MeJA treatment, 225 μmol x L(-1) MeJA and 4 d treatment were optimal. The effective compound contents in callus were obviously higher than in field-grown plants. Therefore, bioreactor culture is efficient for obtaining mass effective compounds of Rhodiolasachalinensis by culturing calluses. This method could provide an alternative material source for production of Rhodiolasachalinensis products.

Acetates ; pharmacology ; Biomass ; Bioreactors ; Cyclopentanes ; pharmacology ; Glucosides ; metabolism ; Oxylipins ; pharmacology ; Peroxidase ; metabolism ; Phenols ; metabolism ; Polysaccharides ; metabolism ; Rhodiola ; metabolism ; Superoxide Dismutase ; metabolism

OBJECTIVETo cultivate adventitious roots of Hypericum perforatum in bioreactors, in order to seek for suitable conditions for adventitious growth.

METHODThe effect of IBA concentration, sugar type and concentration, inoculum volume and air volume of adventitious roots on the cultivation of adventitious roots of H. perforatum was observed in a 5 L air-lift bioreactor.

RESULTAdventitious roots of H. perforatum were cultivated in a MS culture dish. With the increase of IBA concentration, the propagation coefficient of adventitious roots of H. perforatum was on the rise. The IBA concentration ranging between 1.25-1.75 mg x L(-1) was suitable for the growth of adventitious roots. Adventitious roots grew best with sucrose in MS medium, with the propagation coefficient up to 22.15. When sucrose concentration was 30 g x L(-1), fresh weight, dry weight and propagation coefficient reached the maximum value. An adventitious root reactor with an inoculum volume of 20 g was favorable for the growth of adventitious roots. The air volume of reactors of 0.075 vvm (air volume/culture volume per minute) was favorable for the growth of adventitious roots, with the significant increase in the propagation coefficient of adventitious roots. In the amplification experiment, we found that the cultivation conditions of adventitious roots in a 5 L bioreactor was completely applicable to that in 10 and 20 L bioreactors, and adventitious roots grew well in a large bioreactor.

CONCLUSIONIBA concentration, sugar type and concentration, inoculum volume and air volume had a significant effect on the growth of adventitious roots.

Air ; Biomass ; Bioreactors ; Carbohydrates ; pharmacology ; Dose-Response Relationship, Drug ; Hypericum ; drug effects ; growth & development ; Indoles ; pharmacology ; Plant Roots ; drug effects ; growth & development ; Sucrose ; pharmacology ; Tissue Culture Techniques ; instrumentation ; methods

OBJECTIVETo explore the factors affecting the growth of protocorms of Dendrobium candidum and substance synthesis in a reactor, in order to provide a new method for mass production of raw materials of D. candidum.

METHODProtocorms in vitro were used as experimental materials to study the effect of inoculum volume, light intensity and air volume on the growth of protocorms of D. candidum and the accumulation of polysaccharide and dendrobine in a 3 L-air lift balloon type bioreactor.

RESULTAfter 30 days of cultivation in a bioreactor, protocorms became dark green and grew well at the inoculum volume of 10 g x L(-1). The polysaccharide content in protocorms showed no difference at various inoculum volumes; whereas the dendrobine content showed differences (with the highest treatment at the inoculum volume of 10 g x L(-1)), particularly the productions of polysaccharide and alkaloid reached the maximum at the inoculum volume of 10 g x L(-1). The condition of 1 600 lx of light intensity was the most favorable for the growth of protocorms. Though light played a role of improving the accumulation of polysaccharide in protocorms of D. candidum, it could inhibit the accumulation of dendrobine. Polysaccharide content and production were better under light conditions of 1 600 and 2 400 lx than dark conditions. Despite the maximum dendrobine content in dark conditions, the dendrobine production showed the maximum in the light condition of 1 600 lx due to poor growth of protocorms. Protocorms grew well and became dark green at the air volume of 0.2 vvm (air volume culture volume per minute) , which was better than at 0.1 and 0.3, with maximum polysaccharide and dendrobine contents and productions.

CONCLUSIONIn a 3 L-air lift balloon type bioreactor with a working volume of 2 L, the conditions of 10 inoculum volume, 1 600 lx light intensity and 0.2 air volume were favorable for the growth of protocorms and the production of dendrobine. This demonstrates that the cultivation of D. candidum and substance synthesis in a reactor is an effectie approach for mass production of polysaccharide and dendrobine.

Air ; Alkaloids ; metabolism ; Bioreactors ; Dendrobium ; growth & development ; metabolism ; radiation effects ; Dose-Response Relationship, Radiation ; Light ; Plants, Medicinal ; growth & development ; metabolism ; radiation effects ; Polysaccharides ; metabolism ; Tissue Culture Techniques ; methods

The apoptotic effects of shikonin (5,8-dihydroxy-2-[(1R)-1-hydroxy-4-methylpent-3-enyl]naphthalene-1,4-dione) on the human colon cancer cell line SNU-407 were investigated in this study. Shikonin showed dose-dependent cytotoxic activity against SNU-407 cells, with an estimated IC50 value of 3 µM after 48 h of treatment. Shikonin induced apoptosis, as evidenced by apoptotic body formation, sub-G1 phase cells, and DNA fragmentation. Shikonin induced apoptotic cell death by activating mitogen-activated protein kinase family members, and the apoptotic process was mediated by the activation of endoplasmic reticulum (ER) stress, leading to activation of the PERK/elF2α/CHOP apoptotic pathway, and mitochondrial Ca2+ accumulation. Shikonin increased mitochondrial membrane depolarization and altered the levels of apoptosis-related proteins, with a decrease in B cell lymphoma (Bcl)-2 and an increase in Bcl-2-associated X protein, and subsequently, increased expression of cleaved forms of caspase-9 and -3. Taken together, we suggest that these mechanisms, including MAPK signaling and the ER-and mitochondria-mediated pathways, may underlie shikonin-induced apoptosis related to its anticancer effect.
Apoptosis ; bcl-2-Associated X Protein ; Caspase 9 ; Cell Death* ; Cell Line ; Colon* ; Colonic Neoplasms* ; DNA Fragmentation ; Endoplasmic Reticulum* ; Extracellular Vesicles ; Humans* ; Inhibitory Concentration 50 ; Lymphoma, B-Cell ; Mitochondria ; Mitochondrial Membranes ; Protein Kinases

BACKGROUND: Reactive oxygen species (ROS) are involved in various cellular diseases. Excessive ROS can cause intracellular oxidative stress, resulting in a calcium imbalance and even aging. In this study, we evaluated the protective effect of esculetin on oxidative stress-induced aging in human HaCaT keratinocytes. METHODS: Human keratinocytes were pretreated with esculetin for 30 minutes and treated with H₂O₂. Then, the protective effects on oxidative stress-induced matrix metalloproteinase (MMP)-1 were detected by Flou-4-AM staining, reverse transcription-PCR, Western blotting, and quantitative fluorescence assay. RESULTS: Esculetin prevented H₂O₂-induced aging by inhibiting MMP-1 mRNA, protein, and activity levels. In addition, esculetin decreased abnormal levels of phospho-MEK1, phospho-ERK1/2, phospho-SEK1, phospho-JNK1/2, c-Fos, and phospho-c-Jun and inhibited activator protein 1 binding activity. CONCLUSIONS: Esculetin prevented excessive levels of intracellular calcium and reduced the expression levels of aging-related proteins.
Aging ; Blotting, Western ; Calcium ; Fluorescence ; Humans ; Hydrogen Peroxide ; Hydrogen ; Keratinocytes ; Matrix Metalloproteinase 1 ; Oxidative Stress ; Reactive Oxygen Species ; RNA, Messenger ; Skin ; Transcription Factor AP-1

Background/Aims: Accumulating evidence indicates that L-carnitine (LC) protects against multiorgan damage through its antioxidant properties and preservation of the mitochondria. Little information is available about the effects of LC on renal fibrosis. This study examined whether LC treatment would provide renoprotection in a rat model of unilateral ureteral obstruction (UUO) and in vitro. Methods: Sprague-Dawley rats that underwent UUO were treated daily with LC for 7 or 14 days. The influence of LC on renal injury caused by UUO was evaluated by histopathology, and analysis of gene expression, oxidative stress, mitochondrial function, programmed cell death, and phosphatidylinositol 3-kinase (PI3K)/ AKT/forkhead box protein O 1a (FoxO1a) signaling. In addition, H2O2-exposed human kidney cells (HK-2) were treated with LC. Results: LC treatment inhibited expression of proinflammatory and profibrotic cytokines, and was followed by a significant attenuation of tubulointerstitial inflammation and fibrosis. The increased oxidative stress caused by UUO was associated with mitochondrial dysfunction and excessive apoptosis and autophagy via PI3K/AKT/FoxO1a-dependent signaling, and this was abrogated by administration of LC. In H2O2-exposed HK-2 cells, LC decreased intracellular production of reactive oxygen species, and suppressed expression of profibrotic cytokines and reduced the number of apoptotic cells. Conclusions LC protects against the progression of tubulointerstitial fibrosis in an obstructed kidney.

Few studies have evaluated the role of autophagy in the development of oxaliplatin (OXT) resistance in colon cancer cells. In this study, we compared the role of autophagy between SNU-C5 colon cancer cells and OXT-resistant SNU-C5 (SNU-C5/OXTR) cells. At the same concentration of OXT, the cytotoxicity of OXT or apoptosis was significantly reduced in SNU-C5/OXTR cells compared with that in SNU-C5 cells. Compared with SNU-C5 cells, SNU-C5/OXTR cells exhibited low levels of autophagy. The expression level of important autophagy proteins, such as autophagy-related protein 5 (Atg5), beclin-1, Atg7, microtubule-associated proteins 1A/1B light chain 3B I (LC3-I), and LC3-II, was significantly lower in SNU-C5/OXTR cells than that in SNU-C5 cells. The expression level of the autophagy-essential protein p62 was also lower in SNU-C5/OXTR cells than in SNU-C5 cells. In SNUC5/OXTR cells, the production of intracellular reactive oxygen species (ROS) was significantly higher than that in SNU-C5 cells, and treatment with the ROS scavenger N-acetylcysteine restored the reduced autophagy levels. Furthermore, the expression of antioxidant-related nuclear factor erythroid 2-related factor 2 transcription factor, heme oxygenase-1, and Cu/Zn superoxide dismutase were also significantly increased in SNU-C5/OXTR cells. These findings suggest that autophagy is significantly reduced in SNU-C5/OXTR cells compared with SNU-C5 cells, which may be related to the production of ROS in OXT-resistant cells.

Background/Aims: Cigarette smoking is an important modifiable risk factor in kidney disease progression. However, the underlying mechanisms for this are lacking. This study aimed to assess whether nicotine (NIC), a major toxic component of cigarette smoking, would exacerbates tacrolimus (TAC)-induced renal injury. Methods: Sprague-Dawley rats were treated daily with NIC, TAC, or both drugs for 4 weeks. The influence of NIC on TAC-caused renal injury was examined via renal function, histopathology, oxidative stress, mitochondria, endoplasmic reticulum (ER) stress, and programmed cell death (apoptosis and autophagy). Results: Both NIC and TAC significantly impaired renal function and histopathology, while combined NIC and TAC treatment aggravated these parameters beyond the effects of either alone. Increased oxidative stress, ER stress, mitochondrial dysfunction, proinf lammatory and profibrotic cytokine expressions, and programmed cell death from either NIC or TAC were also aggravated by the two combined. Conclusions Our observations suggest that NIC exacerbates chronic TAC nephrotoxicity, implying that smoking cessation may be beneficial for transplant smokers taking TAC.