KR100313347B1 - Use of Panax ginsenoside as egg promoter - Google Patents

Abstract

The present invention processes the ginsenosides (ginsenoside) isolated from ginseng in the egg Ca ²⁺ - active chroman fluoride channel (Ca ²⁺ -activated chloride channels) because writing enable the method to promote oocyte maturation of the oocyte maturation and the intrinsic The present invention relates to the use of cenosides as an oocyte maturation accelerator. The treatment of ginsenosides isolated from ginseng on frog eggs and the activity of Ca ²⁺ -activated chromide channels with varying voltage were carried out. The experiment was repeated to examine the relationship between Ca ²⁺ -activated chromide channel activation and ginsenosides. As a result, the ginsenosides isolated from ginseng bind to the ginsenoside binding protein present in the egg cell membranes, resulting in phospholipase. activate the C and activated phospholipase C is sikimyeo release the membrane from the IP ₃ is IP ₃ is calcium It acts on the IP ₃ binding site in the reservoir to release calcium from the calcium reservoir into the cytoplasm, which in turn activates the Ca ²⁺ -activated chromide channel, which has an excellent effect of promoting oocyte maturation.

Description

Use of ginseng ginsenosides as an oocyte maturation accelerator

The present invention relates to the use of ginseng ginsenosides as an oocyte maturation accelerator. More specifically, the present invention relates to promoting oocyte maturation of ginseng ginsenosides by activating the Ca ²⁺ -activated chromide channel present in the egg to induce maturation of the egg.

Frog eggs (Xenopus laevis oocytes) are widely used to encode exogenously injected RNA or DNA into cell membrane proteins. In particular, electrophysiological techniques have been used to study or characterize the receptors, transporters and ion channels expressed in frog eggs (Dascal, Crit. Rev. Biochem. Mol. Biol. 22 , 317-387, 1987; Snutch TINS, 11, 250-256, 1988).

On the other hand, it has been reported that some types of endogenous ion channels exist in frog eggs. Double well-known that there are many Ca ²⁺ - there may be mentioned ^{- (channels Ca 2+ -activated Cl)} (Barish, J. Physiol 342, 309-325, 1983 active chroman fluoride channel; Takashi et al,.. Proc. Natl. Acad. Sci. 84, 5063-5067, 1987). The physiological function of this channel is unknown, but it is known to play an important role in fertilization potential change and anionic conductance during fertilization of sperm and eggs in mature eggs (Webb and Nucitelli, Dev. Biol. 107, 395-406, 1985). Ca ²⁺ - are known to be very sensitive to inhibition by active chroman fluoride channel broker ^{(Ca 2+ activated chloride channel blocker)} (White and Aylwin, - active chroman fluoride current nip rumsan (niflumic acid), such as Ca ²⁺ Mol. Pharmacol. 37, 720-724, 199O). On the other hand, acetylcholine, known as a neurotransmitter, is known to activate Ca ²⁺ -activated chromide channels by acting on muscarinic receptors that are endogenous in these oocytes (Moriarty et al. , Proc. Natl. Acad. Sci. 85, 8865-8869, 1988). Acetylcholine acts on the muscarinic receptor to activate phospholipase C, resulting in the production of inositol-triphosphate (IP ₃ ), now known as a second messenger and, the generated IP ₃ is the free calcium mobilization (free Ca ²⁺⁾ into the cytoplasm seeker Ca ²⁺ - been found to enable the active chroman fluoride channel. This effect by acetylcholine is known to promote or promote the maturation of egg cells by progesterone (Dascal, Crit. Rev. Biochem. Mol. Biol. 22, 317-387 1987).

Interestingly, a study demonstrating the inclusion of an enzyme called phospholipase C in the activation of Ca ²⁺ -activated chromide channels, in fact the direct injection of free Ca ²⁺ or IP _{3 into} the cytoplasm. Injection is widely known to activate Ca ²⁺ -activated chromide channels (Miledi and Parker, J. Physiol. 357, 173-183, 1984; Hartzell, J. Gen. Physiol. 108, 157-175, 1996). Ca ²⁺ -activated chloride channels are known to exist in the nervous system in addition to frog eggs (Owen et al, Nature, 311, 567-569, 1984; Lascola and Kraig, J. Neurosci 16, 2532-2545, 1996).

Examples of other fluoride croissant channel (chloride channel) present in the frog oocytes Ca ²⁺ that is activated when the hyperpolarization (hyperpolarization) the egg - it is reported that there is no response croissant fluoride channel (Ca ²⁺ -insensitive chloride channel). This channel is not inhibited by niflumic acid but has been reported to be inhibited by substances such as barium or stilbene (Kowdley et al, J. Gen. Physiol. 103, 217 -230, 1994). In addition, the chloride and starch-activated channels included in osmoregulatory are used for trivalent cations such as gadolinium and lanthanum. It is reported to be inhibited (Ackerman et al, J. Gen. Physiol. 103, 153-179, 1994).

Ginsenoside is widely known as a physiological or pharmacologically active ingredient of ginseng (Nah, Korean J Ginseng Sci. 21, 1-12, 1997; Nah, Curr Tr Neurosc. 6, 17-30, 1998). Ginsenosides have been shown to exert a variety of efficacy through in vivo or in vitro studies (Nah, Korean J Ginseng Sci. 21, 1-12, 1997). However, research on increasing reproduction or reproduction by ginseng ginsenosides is very small. However, there are reports of clinical cases on the improvement of spermtozoa motility and the promotion of spernntogenesis in infertile men (Shinshin tea, clinical effect of Korean ginseng, Vol. 2, 31-34, 1994). ). On the other hand, in menopausal women, it has been reported that administration of ginseng may improve ovarian function by increasing blood inflow into the ovarian parenchyma with decreased function and by promoting the production of estrogen, a major ovarian hormone (Etrogen). Clinical Effects of Korean Ginseng, Vol. 1, 157-163, 1993).

On the other hand, frog eggs (Xenopus laevis oocytes) are widely available as models for the development, maturation and ovulation of eggs because they are readily available in large quantities and larger in size than those of other animal species (Maller and Krebs, Curr. Topics Cell Regul. 16, 271-311). The hormone involved in the maturation of frog eggs has been reported to be progesterone, which binds to the progesterone receptors on the plasma membrane of the egg and activates phospholipase C. Induction of egg maturation (Stith et al., Cell Calcium 13, 341-352, 1992).

However, there is no research on the effects of ginsenosides directly on oocytes present in the female genitalia. The present inventor is a ginsenoside effective component of ginseng Ca ²⁺ present in egg - it was found that the newly sikindaneun enable active chroman fluoride channel (Ca ²⁺ -activated chloride channels). Ginsenosides in the binary process is phospholipase C (phospholipase C) free Ca ²⁺ from the train is activated as a result of IP ₃ generation activate the enzyme of the enzyme cytosolic intracellular calcium stores (Ca ²⁺ pools) of (free Ca ²⁺ ) was stimulated to be released into the cytoplasm and released calcium was found to play an important role in the maturation process of mammalian eggs. Accordingly, the present invention is directed to the treatment of infertility and the development of contraceptive drugs by activating Ca ²⁺ -activated chromide channels present in oocytes present in the female genitalia and promoting or elevating the maturation of egg cells. It is meaningful to use.

Accordingly, an object of the present invention is to provide a use of ginsenosides isolated from ginseng as an oocyte maturation accelerator. Another object of the present invention is to provide a method of promoting the maturation of eggs by activating the Ca ²⁺ -activated chromide channel by treating the egg with ginsenosides isolated from ginseng.

The object of the present invention is to treat ginsenosides isolated from ginseng in frog eggs and to investigate the activity of Ca ²⁺ -activated chromide channels while varying the voltage, and then repeating the experiment while varying the dosage of ginsenosides. by ginsenosides isolated from ginseng Ca ²⁺ - investigate the activation of the active chroman fluoride channel Ca ²⁺ for the pharmacological test of the experiment-and handle active chroman fluoride channel brokers nip rumsan in frog oocytes again Caffeine or 1, 2-bis- (2-aminophenoxy) ethane-NNN`N`tetra-acetic acid (1,2-bis- (2-amino phenoxy) ethane, treated with ginsenosides or damaging calcium reservoirs -NNN`N`-tetra-acetic acid; - after the treatment the active chroman fluoride channel Ca ²⁺ - BAPTA) activated Ca ²⁺ was measured the activity of the active chroman fluoride channel. Subsequently, the action of ginsenosides occurs in the cytoplasm or other extracellular cells. After activation or inactivation of phospholipase C, an enzyme that activates the Ca ²⁺ -activated chromide channel, the action of ginsenosides is observed. To investigate the signaling process of ginsenosides in the activity of Ca ²⁺ -activated chromide channels, and to treat frog eggs with progesterone hormones and ginsenosides, which induce maturation of frog eggs, germ cells vesuicle breakdown (GVBD) was achieved by investigating the oocyte maturation capacity of ginsenosides.

1a when the turn would not process the ginseng ginsenoside basis appears -Ca ²⁺ - a ^{- (currents basal Ca 2+ -activated Cl} ) have shown that the generated according to the number of voltage steps (steps voltage) graph active chroman fluoride current .

1b or Ca when treated with ginseng ginsenoside 10 ㎍ / ㎖ in frog egg²⁺Active chromide current (Ca²⁺-activated Cl- currents) is a graph showing the activation according to the voltage step.

FIG. 1c is a graphical representation of the I-V relationship between FIGS. 1a and 1b.

FIG. 1d is a graph showing the tail-currents confirming that the current activated by ginseng ginsenoside is Ca ²⁺ -active chromide current.

2 shows that Ca ²⁺ -active chromide current increases as ginseng ginsenoside dose is increased.

Third ginseng ginsenoside turning Ca ²⁺ - active chroman fluoride channel (Ca ²⁺ -activated ^C1-channels) the activating effect of the Ca ²⁺ - active chroman fluoride channel broker (Ca ²⁺ -activated Cl ^- channels blocker Is blocked by niflumic acid (NFA).

No. 4a 4b assist the turning of the ginseng ginsenoside Ca ²⁺ - a diagram showing the mechanism of action for activating the active chroman fluoride channel yirueojim to mobilize calcium from the cytoplasm in (endoplasmic reticulum) calcium stores (Ca ²⁺ pool).

The fifth turning ginseng binary Ca ²⁺ by ginsenoside-activating effect of a picture is ^- (channels Ca ²⁺ -activated Cl) represents the yirueojim in the cell membrane (cell membrane) active chroman fluoride channel.

Figure 6 shows that ginseng ginsenosides contain an enzyme called phospholipase C, which acts to activate the Ca ²⁺ -activated chromide channel.

The present invention comprises the steps of preparing a frog egg (Xenopus laevis oocutes) according to the method of Zerr et al. And preparing ginsenosides according to Ando et al. From red ginseng; The prepared frog eggs were treated with or without ginsenosides, and the chromide currents generated for 300ms were recorded by increasing the voltage from -80mv to 20mv, thereby determining the Ca ²⁺ -activated chromide channel activity of ginsenosides. Investigating; Measuring a residual current after investigation of the activity of the Ca ²⁺ -activated chromide channel by the ginsenoside treatment; Measuring the activity of the Ca ²⁺ -activated chromide channel after treatment of the depleted egg cells with various doses of ginsenosides; Niplum acid, a broker of Ca ²⁺ -activated chromide channels, was pretreated in frog eggs and treated with ginsenosides or washed after niprum acid, followed by ginsenosides and the activity of each Ca ²⁺ -activated chromide channel was measured. Doing; Gene activated by Ca ²⁺ ginsenoside-active chroman fluoride was combined with caffeine or calcium to compromise the calcium storage in the channel by processing the known BAPTA as a calcium blocking action again ginsenoside treated Ca ^{2 +} - activated Measuring chromide channel activity; Ca ²⁺ -activated chromide channel activity was measured in each case of ginsenoside treatment in vitro and in the intracellular cytoplasm to determine whether ginsenosides acted on the cell membrane of the oocyte or in the cytoplasm. Investigating; Inactivating or activating phospholipase C, which releases calcium from the calcium reservoir into the cytoplasm to activate the Ca ²⁺ -activated chromide channel, followed by ginsenoside treatment and measuring Ca ²⁺ -activated chromide channel activity. ; Germinal vesicle breakdown, which occurs when oocytes are treated with progesterone, a hormone that induces the maturation of frog eggs, and when oocytes are treated with ginsenosides, or when both progesterone and ginsenosides are treated at the same time; GVBD) to investigate the synergistic effect or promote oocyte maturation of ginsenosides.

In the present invention, two-electrode voltage clamps were performed at room temperature and the amplifier used in the experiment was a Geneclamp 500 (Axon Instruments, CA USA) and the amplifier was connected to a Macintosh Quadra 800 computer. Was used. Linear leak and capacitance currents were modified by the P / 4 leak substration procedure. Data collection and analysis was done according to the Pulse and PulseFit program (Heka, Germany), and the graphs used the IGOR (Wavemetrics) and Kaleidagraph (Synergy software) programs.

Peak currents in Ca ^{2+ -activated} chloride channel currents activated by ginsenosides were collected for statistical significance evaluation, and their expression was taken as mean ± SEM (μA) (Zerr et al., J Neurosci. 18, 2842-2848, 1998). Statistical significance test was confirmed by unpaired Student`st test, and p <0.05 was considered significant.

EXAMPLE

Example 1 Preparation of Frog Eggs and Ginseng Ginsenosides

Frog eggs (Xenopus laevis oocytes) were prepared according to Zerr et al. (Zerr et al., J. Neurosci. 18, 2843-2848, 1998). In other words, mature frog females (Xenopus laevis) were anesthetized in ice, and then intraperitoneally cut and collected by collagenase [(2 mg / mL ND 96 media (96 mM Nacl, 2 mM KCl, 1 mM MgCl2, 1.8). mM CaCl 2, 5 mM Hepes, pH 7.5)] to remove the follicular membrane of the egg, which had been washed three times with ND 96 media. The gene was removed and frog eggs were then tested under perfusion conditions in ND 96. Ginsenosides were dissolved in ND 96 medium (stock solution: 200 μg / mL) and then in ND 96 medium at different doses. Ginseng ginsenosides (ginseng total saponins) were prepared from red ginseng according to the method of Ando et al. (Ando et al., Syoyakukaku Zasshi 25, 28-34, 1971) and used in the experiment.

Example 2 Ca ²⁺ -Activated Chloride Channel Activity Investigation with Ginsenosides Isolated from Ginseng

Ca²⁺The chromide currents generated by the activation of the active chromide channel currents by ginsenosides can be fixed by fixing the frog eggs at -80 mV (ie fixed potential: -80 mV) and then positive voltage step by 20 mV. It is recorded as the chromide current generated by continuous stimulation for 300 ms moving to +60 mV. In other words, the relationship of -60, -40, -20, 0, 20, 40 and 60 mV at the fixed potential and the continuous stimulus of 300 ms is called the current-voltage (I-V) relationship. Experimental results, as shown in Figure 1a when not treated with ginsenoside purely separated from ginseng Ca²⁺Although the activity of the active chromide channel is not large (basal level), as shown in FIG. 1b, when ginsenosides 10 μg / mL are treated in frog eggs, the chromide channel is activated 20 times more than that of the non-treated chromide channel. Chloride Current (outward Cl- current) is shown. In order to further confirm the action of ginsenosides, the relationship between the fluoride current and the voltage (I-V relationship) is illustrated in FIG. 1C. As shown in this figure, the treatment of ginsenosides (10 μg / mL, open circle) yielded some inward chromide currents at -60 mV and -40 mV.^-current, causing an inward Cl current at -30 mV.^-current flows out^-current). Furthermore, as the voltage step was increased, a large amount of outflow chromide current, which was not seen in the basal and closed circles, was generated by the treatment of ginsenosides.

Example 3 Ca ²⁺ -Activated Chloride Channel Activation Assay by Ginsenosides

In this example, the tail-current by ginsenosides was examined to further verify that the chromide channel is activated by the ginsenosides shown in Example 2. This is a way to check the reversal potential of the chromide channel, first depolarizing the frog egg for 400 ms at +40 mV at a fixed potential and then again for voltage steps +20, 0, -20, -40, -60. And 300 ms at -80 mV. As a result, as shown in Figure 1d ginsenoside (10 ㎍ / mL treatment as the voltage moves from +20 mV to -20 mV, that is, the negative potential (outward current) as Reduced but still showing outflow chromide current, and from -40 mV inward Cl ^- current was generated and inflow chromide current was maintained until -80 mV. The mV is reversed, and the IV relationship of Figure 1c also shows that the voltage step at which the inward current turns to the outward current is about -30 mV. Showed activating Ca ²⁺ -activated chromide channel (5 cells used, n = 5).

Example 4 Investigation of Ca ²⁺ -Activated Chloride Channel Activity According to Dose of Ginsenoside

In this example, all the experimental conditions were carried out as in Example 2, and the amount of ginsenosides was 1, 3, 10, 30 µg / mL. As a result, it can be seen that Ca ²⁺ -activated chromide channel is activated at the dose dependency of ginsenoside as shown in FIG. 2. Increasing the dose of ginsenosides to frog eggs increases the activation of Ca ²⁺ -activated chromide channels and has been shown to be nearly saturated at 10 μg / mL of ginsenosides. The results show that the concentration of ginsenosides, ie EC _50, is 4.2 μg / mL to activate the Ca ²⁺ -activated chromide channel. The number of oocytes used was 5 for each concentration (n = 5, mean ± SEM).

Example 5 Pharmacological Assay of Ca ²⁺ -Activated Chloride Channel Activation by Ginsenosides

Dark ginsenosides actually Ca ²⁺ in frog oocytes - You Prum broker drug active chroman channel acid fluoride - to investigate the active chroman fluoride active role solidifying the channel pharmacologically Ca ^2+; was used (niflumic acid NFA) . As shown in FIG. 3, pretreatment with 100 μM NFA followed by ginsenoside (10 μg / mL) with NFA did not activate Ca ²⁺ -activated chromide channel by ginsenoside and also washed NFA. Ginsenosides activated Ca ²⁺ -activated chromide channels when removed and then retreated with ginsenosides. The results show that the action of NFA is in a reversible manner and that ginsenosides activate the Ca ²⁺ -activated chromide channel (n = 5).

[Example 6]: Ca by Ca ^{2+ 2+} - active chroman fluoride channel activating black

In this embodiment, Ca ²⁺ - As shown in the name and characteristics of the active chroman fluoride channel is channel black that was activated by Ca ²⁺ that is, ginsenoside is Ca ²⁺ - activated croissant before fluoride functional group that activates the channel It was tested whether Ca ²⁺ ions were included (Lechleiter and Clapham, Cell 69, 283-294, 1992). Binary processing of the ginsenosides is Ca ²⁺ release by the calcium opening the calcium stores (Ca ²⁺ pools) present in the cytoplasm mangsangche (endoplasmic reticulum) in the egg cell into the cytoplasm - induces activation of the active chroman fluoride channel of egg cells Caffeine (Hulsmann et al., Eur J Physiol 436, 49-55, 1988; Parekh et al., J. Physiol. 469, 653-671 1993), known to destroy calcium reservoirs in advance; and When administered intracellularly, 1,2-bis- (2-aminophenoxy) ethane-NNN`N`-tetra-acetic acid (1,2-bis- () is known to bind intracellular calcium and inhibit the action of calcium. 2-aminophenoxy) ethane-NNN`N`-tetra-acetic acid; BAPTA) (Kavanaugh et al, Biochem. J., 277, 899-9O2. 1991; Hartzell, J. Gen. Physiol. 108, 157-175, 1996). As a result, as shown in FIG. 4A, after activating the Ca ²⁺ -activated chromide channel by ginsenoside (30 µg / mL), pretreatment with 10 mM caffeine for about 5 minutes, followed by the presence of caffeine When the side was treated, the action of ginsenosides was almost blocked. In addition, when ginsenoside (30 µg / mL) was treated 10 minutes after administration of H ₂ 0 or 1 mM BAPTA (final concentration) into the cytoplasm of frog eggs, only when H ₂ 0 was administered as shown in FIG. 4B. Ginsenosides activated Ca ²⁺ -activated chromide channels and were rarely activated when BAPTA was administered. As a result, it can be seen that ginsenosides stimulate calcium to be released from the calcium reservoir into the cytoplasm (n = 7, mean ± SEM).

Example 7 Investigation of Action Location of Ginsenoid

Whether ginsenosides act on the cell membrane or within the cytoplasm in activating Ca ²⁺ -activated chromide channels has not been elucidated. In the present embodiment, first to examine the ginsenoside (30 ㎍ / mL) to the outside of the cell and then confirmed that the Ca ²⁺ -activated chromide channel is activated, a very small amount of nano-injector that can be injected into the cell ( The effects of ginsenosides (1µg / oocyte) were intracellularly administered using nanoinjectors. Experimental results show that intracellular injection of ginsenosides (arrows in FIG. 5 indicate injection of ginsenosides) showed little activation of Ca ²⁺ -activated chromide channels as shown in FIG. 5 (n = 5). ). As a result, it can be seen that ginsenosides are acting extracellularly.

Example 8 Investigation of Signaling Process for Ca ²⁺ -Activated Chloride Channel Active Group of Ginsenosides

In this example, the process of activating Ca ²⁺ -activated chromide channel was investigated. The treatment of ginsenosides in frog eggs so far binds to an unknown yet unknown binding protein (ginsenoside binding protein) present in the egg cell membrane, which then activates the binding protein (or ginsenoside receptor). It produces secondary carriers that act on the calcium reservoir, releasing calcium in the cytoplasm and the released calcium ions eventually activate the Ca ²⁺ -activated chromide channel. However, the signal transduction pathways involved in the process are not yet known. Acetylcholine, a neurotransmitter already known to activate Ca ²⁺ -activated chromide channels, binds to acetylcholine receptors present in frog eggs to activate phospholipase C (PLC) and is produced by PLC activation. IP ₃ is known to activate the Ca ²⁺ -activated chromide channel by binding to the IP ₃ receptor present in the calcium reservoir and releasing calcium from the calcium reservoir into the cytoplasm. Therefore, in this example, it was examined whether Ca ²⁺ -activated chromide channel is activated by ginsenoside activation of PLC. For this study, the action of ginsenosides was investigated using an active PLC inhibitor (U-73122), which inactivates PLC activity, and an inactive PLC inhibitor (U-73343), which is known to rarely inactivate PLC (Thompson et al. , J. Biol. Chem. 266, 23856-23862, 1991; Davletov et al., EMBO J. 17, 39O9-3920, 1998). The results were based on 100% Ca ²⁺ -activated chromide peak current which is activated when only ginsenosides (30 μg / mL) were initially treated, and U-73343 or 5 minutes after ginsenosides were washed and removed. The percentage of recovery of Ca ²⁺ -activated chromide peak current when the same concentration of ginsenosides were treated with U-73122 on the same cells (eg, eggs) was calculated. As a result, as shown in FIG. 6, when treated with 1μM U-73343 (inactive PLC inhibitor), Ca ²⁺ -activated chromide current activated by ginsenoside recovers about 80% when it is not treated. Treatment with 1 μM U-73122 (active PLC inhibitor) significantly reduced the recovery than without treatment (approximately 50%) (Cruzblanca et al, Proc. Natl. Acad. Sci. 95, 7151-7156, 1998) (n = 7-8. Mean ± SEM * p <0.05). According to these results, the treatment of ginsenosides on the ovum cells activates the PLC by binding the ginsenoside to the ginsenoside binding protein (or ginsenoside receptor) present in the cell membrane, and the activated PLC releases IP ₃ from the cell membrane. And the released IP ₃ acts on the IP ₃ binding site in the calcium reservoir to release calcium from the calcium reservoir into the cytoplasm, and the released calcium again activates the Ca ^{2+ -active} chloride channel, indicating that this channel is opened. have.

Example 9 Induction of Germinal vesicle Breakdown by Ginsenosides

Experiments on the maturation of frog oocytes (Xenopus laevis oocytes) by treatment with ginsenosides and progesterone were carried out according to the method of Bandyopadhyay et al. (Bandyopadhyay et al., Gen. comp. Endorinol. 109, 293-301, 1998). Was carried out. In other words, the oocytes collected from mature frogs were treated with collagenase to prepare oocytes (denuded oocytes) that had just been removed. Ten eggs were prepared in a 24-well culture dish. Determination of what happens when the egg matures after treatment with progesterone 1nM, 10nM, 10OnM and 1000nM and ginsenosides 50, 100, 250 and 50Og / mL in each well containing the egg for 24 hours Germinal vesicle breakdown (GVBD) was used as a measure. As a result, as shown in Table 1, almost no GVBD occurred in the 1nM progesterone-treated group, which was used as a control group, which means that no egg maturation occurred, and about 25% of GVBD occurred in the 10nM progesterone-treated group. did. In the 100 nM progesterone treatment group, about 71% of GVBDs occurred. Almost GVBD occurred in the 1000 nM progesterone alone group. Treatment with ginsenosides of 100 μg / mL produced 7% GBVD, but treatment of more than 250 μg / mL resulted in more than 50% of GVBD. Treatment with more than 500 μg / mL showed 75% GVBD. In other words, 10% of progesterone-treated group developed about 25% of GVBD, but when treated with 100µg / mL, 74% of GVBD occurred. As such, the treatment of ginsenosides has been shown to promote GVBD induction by progesterone. As a result, when progesterone and ginsenosides are treated together, it can be seen that they promote or increase the maturation of eggs, which suggests that acetylcholine promotes the proliferation of eggs by progesterone. (Dascal et al., J. Exp. Zool. 230, 131-140, 1984; Dascal, et al., Gamete Res. 12, 171-178, 1985).

TABLE 1

Effect of Ginsenosides on Oocyte Maturation

In the present invention, as described in the above embodiment, ginsenosides isolated from ginseng bind to ginsenoside binding proteins present in the egg cell membranes of mammals to activate phospholipase C, and activated phospholipase C represents IP ₃ . It releases from the cell membrane and IP ₃ acts on the IP ₃ binding site in the calcium reservoir to release calcium from the calcium reservoir into the cytoplasm, which in turn activates the Ca ²⁺ -activated chromide channel to promote oocyte maturation. Since it is effective, it is a very useful invention in the biopharmaceutical industry for infertility treatment and contraceptive development.

Claims (3)

An oocyte maturation accelerator, comprising ginsenosides isolated from ginseng as active ingredients. The oocyte maturation promoter of claim 1, wherein the ginsenoside activates a Ca ²⁺ -activated chromide channel. The oocyte maturation promoter according to claim 1, wherein the ginsenoside activates phospholipase C.