BACKGROUND: Kinetin is a plant hormone that regulates growth and differentiation. Keratinocytes, the basic building blocks of the epidermis, function in maintaining the skin barrier. OBJECTIVE: We examined whether kinetin induces skin barrier functions in vitro and in vivo. METHODS: To evaluate the efficacy of kinetin at the cellular level, expression of keratinocyte differentiation markers was assessed. Moreover, we examined the clinical efficacy of kinetin by evaluating skin moisture, transepidermal water loss (TEWL), and skin surface roughness in patients who used kinetin-containing cream. We performed quantitative real-time polymerase chain reaction to measure the expression of keratinocyte differentiation markers in HaCaT cells following treatment. A clinical trial was performed to assess skin moisture, TEWL, and evenness of skin texture in subjects who used kinetin-containing cream for 4 weeks. RESULTS: Kinetin increased involucrin, and keratin 1 mRNA in HaCaT cells. Moreover, use of a kinetin-containing cream improved skin moisture and TEWL while decreasing roughness of skin texture. CONCLUSION: Kinetin induced the expression of keratinocyte differentiation markers, suggesting that it may affect differentiation to improve skin moisture content, TEWL, and other signs of skin aging. Therefore, kinetin is a potential new component for use in cosmetics as an anti-aging agent that improves the barrier function of skin.
Antigens, Differentiation* ; Cell Culture Techniques ; Epidermis ; Humans ; In Vitro Techniques ; Keratin-1 ; Keratinocytes* ; Kinetin* ; Plants ; Real-Time Polymerase Chain Reaction ; RNA, Messenger ; Skin Aging ; Skin* ; Treatment Outcome ; Water

Liver fibrosis is an important health problem that can further progress into cirrhosis or liver cancer, and result in significant morbidity and mortality. Inhibiting proliferation and inducing apoptosis of hepatic stellate cells (HSCs) may be the key point to reverse liver fibrosis. At present, anti-fibrosis drugs are rare. Kinetin is a type of plant-derived cytokinin which has been reported to control differentiation and induce apoptosis of human cells. In this study, the HSCs were incubated with different concentrations of kinetin. The proliferation of rat HSCs was measured by MTT assay, cell cycle and apoptosis were analyzed by flow cytometry, and the apoptosis was examined by TUNEL method. The expression of Bcl-2 and Bax proteins was detected by immunocytochemistry staining. It was found that kinetin could markedly inhibit proliferation of HSCs. In a concentration range of 2 to 8 μg/mL, the inhibitory effects of kinetin on proliferation of HSCs were increased with the increased concentration and the extension of time (P < 0.01). Flow cytometry indicated that kinetin could inhibit the DNA synthesis from G0/G1 to S phase in a dose-dependent manner (P < 0.01). The apoptosis rates of the HSCs treated with 8, 4 and 2 μg/mL kinetin (25.62% ± 2.21%, 15.31% ± 1.9% and 6.18% ± 1.23%, respectively) were increased significantly compared with the control group (3.81% ± 0.93%) (P < 0.01). All the DNA frequency histogram in kinetin-treated groups showed obvious hypodiploid peak (sub-G1 peak), and with the increase of kinetin concentrations, the apoptosis rate of HSCs also showed a trend of increase. It was also found that kinetin could down-regulate the expression of Bcl-2, and up-regulate the expression of Bax, leading to the decreased ratio of Bcl-2/Bax significantly. The kinetin-induced apoptosis of HSCs was positively correlated with the expression of Bax, and negatively with the expression of Bcl-2. It was concluded that kinetin can inhibit activation and proliferation of HSCs by interrupting the cell cycle at G1/S restriction point and inducing apoptosis of HSCs via reducing the ratio of Bcl-2/Bax.
Animals ; Apoptosis ; drug effects ; Cell Line, Transformed ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; G1 Phase Cell Cycle Checkpoints ; drug effects ; genetics ; Gene Expression Regulation ; Growth Inhibitors ; pharmacology ; Hepatic Stellate Cells ; cytology ; drug effects ; metabolism ; Kinetin ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; antagonists & inhibitors ; genetics ; metabolism ; Rats ; Signal Transduction ; bcl-2-Associated X Protein ; agonists ; genetics ; metabolism

Kinetin (Kn) is a cytokinin growth factor that exerts several anti-aging and antioxidant effects on cells and organs. To investigate the mechanism underlying apoptotic events in aging cells induced by D-galactose (D-gal), we examined the effect of Kn delivered via nuchal subcutaneous injection on D-gal-induced aging and apoptosis in rats. Our results showed that interleukin (IL)-2 levels and mitochondrial membrane potential (DeltaPsim) were decreased by Kn in aging rats while IL-6 production and apoptosis increased. In addition, the expression of anti-apoptotic Bcl-2 was low while that of Bax was high in the aging group. After treated with Kn, compared with aging group, there showed obvious difference in Kn group with elevated IL-2, proliferation index, Bcl-2, DeltaPsim and decreased IL-6 and Bax in splenic lymphocyte. Based on these results, we concluded that Kn can effectively protect the rat spleen from aging, apoptosis, and atrophy.
Aging/drug effects/physiology ; Animals ; Apoptosis/drug effects/*physiology ; Female ; Galactose/*pharmacology ; Interleukin-6/physiology ; Interleukins/physiology ; Kinetin/pharmacology/*physiology ; Male ; Membrane Potential, Mitochondrial/drug effects/physiology ; Rats ; Spleen/*cytology/drug effects/physiology

In order to study the effect of phytohormone on growth and isoflavones contents of Pueraria phaseoloides hairy roots, we cultured the hairy roots with different concentrations of 6-benzylaminopurine (6-BA) alone or in combination with α-naphthaleneacetic acid (NAA). Then we determined the effects of 6-BA alone or in combination with NAA on the growth and the contents of isoflavones compounds and levels of antioxidase activities of hairy roots by spectrophotometry. The results show that 6-BA inhibited the growth, and decreased biomass and total isoflavones compounds of P. phaseoloides hairy roots. Furthermore, the inhibition was increased with the concentrations of 6-BA. Compared with the controls, different concentrations of 6-BA in combination with NAA 2.0 mg/L could inhibit the growth of hairy roots and decrease the content of total isoflavone compounds, and also significantly enhanced the contents of soluble protein and levels of peroxidase (POD) activities, but decreased the activities of superoxide dismutase (SOD). DNA ladders detected by agarose gel electrophoresis can be observed after hairy roots of P. phaseoloides were cultured with 6-BA alone for 30 days, but can appear on the 20th day after culture with 6-BA in combination with NAA 2.0 mg/L. This result indicates that 6-BA or 6-BA in combination with NAA can both stimulate appearance of programmed cell death (PCD), and NAA may play a synergistic role on PCD.
Benzyl Compounds ; Isoflavones ; chemistry ; Kinetin ; pharmacology ; Naphthaleneacetic Acids ; pharmacology ; Plant Growth Regulators ; pharmacology ; Plant Roots ; chemistry ; drug effects ; growth & development ; Pueraria ; drug effects ; growth & development ; Purines

The purpose of this paper is to study the effect of kinetin (Kn) on immunity and splenic lymphocyte proliferation in vitro of aging rats induced by D-galactose (D-gal). Fifty SD rats were randomly divided into five groups: control group, aging model group, Kn low dose group, Kn middle dose group and Kn high dose group. The aging model group was proposed by napes subcutaneous injection of D-gal (125 mg/kg) for 45 d, and anti-aging groups were intragastrically administered with 5, 10, 20 mg/kg of Kn respectively from day 11. IgG, IgA, IgM contents of serum, the apoptosis percentage, stimulation index (SI) and proliferation index (PI) of splenic lymphocyte in vitro were evaluated. The results showed that the apoptosis percentage of splenic lymphocyte in aging model rats was higher, the serum IgG, IgA and IgM contents, SI and PI were lower than control group. Kn significantly decreased the apoptosis percentage of splenic lymphocyte, while increased the serum IgG, IgA and IgM contents, SI and PI in aging model group. These results suggest that Kn could inhibit the apoptosis, while promote the proliferation of splenic lymphocyte, and then effectively enhance the immune power of the aging rats and slow down the aging process.
Aging ; drug effects ; immunology ; Animals ; Antibodies ; blood ; Apoptosis ; Cell Proliferation ; drug effects ; Galactose ; adverse effects ; Kinetin ; pharmacology ; Lymphocytes ; cytology ; Rats ; Rats, Sprague-Dawley ; Spleen ; cytology

The present study was to investigate the effect of kinetin on ovary and uterus of D-galactose-induced female mouse model of aging. Aging female mice model caused by D-galactose were used as model group, the aging model mice intragastrically administered with kinetin solution (daily 25 mg/kg or 50 mg/kg) were used as kinetin groups, and the mice with solvent as normal group (n = 20). To detect the effects of kinetin, estrous cycle, estradiol content, ovarian and uterine wet weight and organ index, SOD and GSH-Px activities, MDA and total protein contents, as well as the reserve function of ovaries were examined. The results showed that, kinetin-induced changes in two kinetin groups were observed, compared with the model group: (1) the estrous cycle was shortened; (2) serum estradiol content was significantly increased; (3) the wet weights of the ovary and uterus were increased significantly; (4) SOD and GSH-Px activities of ovary and uterus were significantly higher; (5) the MDA contents of the ovary and uterus were reduced significantly; (6) total protein contents of the ovary and uterus were increased significantly; (7) the numbers of mature oocytes in fallopian tubes were increased significantly. The results show that kinetin can protect ovary and uterus against oxidative damage, prevent low estrogen secretion caused by ovarian oxidative damage, shorten the estrous cycle in mice, and eventually maintain ovarian and uterine vitalities.
Aging ; Animals ; Estradiol ; metabolism ; Estrous Cycle ; drug effects ; Female ; Galactose ; Kinetin ; pharmacology ; Mice ; Organ Size ; Ovary ; drug effects ; Uterus ; drug effects

OBJECTIVETo study the effect of sucrose and plant growth substances of different concentrations on the induction of test-tube tuberous roots of Rehmannia glutinosa, in order to establish an efficient system for the induction of test-tube tuberous roots from leaves of R. glutinosa.

METHODLeaves from test-tube seedlings of 85-5 R. glutinosa were used as explants. After rooting induction, they were transferred to medium with orthogonal design for inducing test-tube tuberous roots of R. glutinosa.

RESULT AND CONCLUSIONNAA played a significant role in induction of test-tube tuberous roots of R. glutinosa, followed by sucrose and 6-BA. With leaves from test-tube seedlings as the explants, the optimal medium for inducing test-tube tuberous roots of R. glutinosa was MS + BA 3.0 mg x L(-1) + NAA 0.1 mg x L(-1) + sucrose 7%. The study provides an efficient induction system for studies on artificial seeds and secondary metabolism with test-tube tuberous roots of R. glutinosa.

Benzyl Compounds ; Dose-Response Relationship, Drug ; Kinetin ; pharmacology ; Naphthaleneacetic Acids ; pharmacology ; Plant Growth Regulators ; pharmacology ; Plant Leaves ; drug effects ; growth & development ; Plant Roots ; drug effects ; growth & development ; Purines ; Rehmannia ; drug effects ; growth & development ; Seedlings ; drug effects ; growth & development ; Sucrose ; pharmacology ; Tissue Culture Techniques ; instrumentation ; methods

OBJECTIVETo determine the content of baicalin in Scutellaria baicalensis callus induced by different doncentrations of exogenous hormones.

METHODHPLC system was adopted to determine baicalin in S. baicalensis callus. Chromatographic conditions: ODS column was adopted, with methanol-water-phosphate (47: 53: 0.2) as the mobile phase. The flow velocity was 1 mL x min(-1), the detective wavelength was 280 nm, and the temperature of column was room temperature.

RESULTS. baicalensis callus induced by 6-BA 1.0 mg x L(-1) + NAA 0.5 mg x L(-1) showed the highest baicalin content, up to 49.78 mg x g(-1).

CONCLUSIONThe experiment is such a simple, rapid and stable method for determining the baicalin content that it can be used for determining the baicalin content in S. baicalensis callus.

2,4-Dichlorophenoxyacetic Acid ; pharmacology ; Benzyl Compounds ; Chromatography, High Pressure Liquid ; Dose-Response Relationship, Drug ; Flavonoids ; metabolism ; Kinetin ; pharmacology ; Naphthaleneacetic Acids ; pharmacology ; Plant Growth Regulators ; pharmacology ; Purines ; Scutellaria baicalensis ; drug effects ; metabolism ; Tissue Culture Techniques ; methods

Characteristics of fructification and seed ripening of Panax stipuleanatus were studied in this paper with a view to better understand the species' biological characteristics and promote cultivation. Dynamic of fructification and types of fruits were observed in a period of 80 days. The seeds' biological characteristics were evaluated by observing embryo's vitality and changes in form in a period of 225 d. The findings and conclusions were as follows: A plant produces an average of (38.7 +/- 6.5) seeds; Seeds become fully mature 80 days after setting fruit, and fruits grow more rapidly in 0-40 days with an increase of 1 500.83% in fresh weight. Three types of fruits were found, that is mono-seed type, bi-seeds type and triple-seeds type, and the proportion is 9.53%, 58.66%, 1.81%, respectively. The seeds are (5.16-6. 52) mm in length and (3.12-4.95) mm in width. The weight of 1000 seeds is (0.648 +/- 0.014) g with a moisture content of (55.23 +/- 0.49)%. The after-ripening of embryo in Panax stipuleanatus is morphological form, embryos develop 75 days after the seeds' detaching from mother plant and become mature in 165-180 days with the ending of the process of after-ripening. The vitality of embryo reaches 70% , and the length of embryo is (3.217 +/- 0.406) mm and the width (0.872 +/- 0.165) mm. The seeds will germinate in 195 d.
Fruit ; growth & development ; Germination ; physiology ; Kinetin ; biosynthesis ; metabolism ; physiology ; Panax ; chemistry ; physiology ; Physiological Phenomena ; Seeds ; physiology

OBJECTIVETo study the effect of the different constitutions of plant hormone on the development of Anoectochilus roxburghii.

METHODA. roxburghii were harvested after having been cultured for 60 days. An orthogonal design was used to study the effect of NAA and 6-BA on the leaf number, eustipe number, lateral branch number of the stem tip and stem section, and the height of the stem tips. All of the data were processed by SPSS.

RESULT AND CONCLUSIONIt is reported for the first time that NAA could make different development of A. roxburghii at low concentration ( < 1 mg L(-1)) and high concentration ( > 1 mg L(-1)). The optimum constitution of MS medium was NAA 0.5 mg L(-1) + 6-BA 1 mg L(-1) for the growth of the stem tip of A. roxburghii, and NAA 1 mg L(-1) + 6-BA 2 mg L(-1) for the differentiation of bud and the formation of lateral branch of the stem section. The different concentrations of NAA and 6-BA had different effects on the growth and differentiation of the stem tip and the stem section of A. roxburghii.

Benzyl Compounds ; Data Interpretation, Statistical ; Kinetin ; metabolism ; Naphthaleneacetic Acids ; metabolism ; Orchidaceae ; growth & development ; metabolism ; Plant Growth Regulators ; metabolism ; Purines ; Software ; Tissue Culture Techniques