TWI602582B - A use of an extract of cassiopea andromeda - Google Patents

Description

Use of Jellyfish Jellyfish Extract

The present invention relates to the use of a jellyfish jelly extract, and more particularly to the use of a jellyfish jelly extract for the preparation of a formulation for promoting skin cell renewal.

In general, healthy skin will last for 28 days. However, with age, or by external environmental stimuli, the speed of skin renewal will slow down, making the skin rough or dull, and even other Skin problems, so in order to maintain a good skin condition, it is imperative to promote skin cell renewal.

Accordingly, the present invention provides a use of a jellyfish jelly extract which is applied to a preparation for promoting skin cell renewal by extracting an active ingredient extracted from the jellyfish jellyfish.

The use of the extract of the jellyfish jelly of the present invention is for preparing a preparation for promoting skin cell regeneration, wherein the extract of the jellyfish jelly is obtained by a method comprising the steps of: providing a sample of a jellyfish jelly; After the jellyfish sample is homogenized with the monophosphate buffered saline, a supernatant is obtained; the total protein of the jellyfish sample is precipitated by a saturated ammonium sulfate solution (saturated percentage of 60-100%), and a precipitate is obtained. The precipitate was dissolved in 50 mM Tris-HCl buffer; and dialyzed against the Tris-HCl buffer to remove small molecular substances having a molecular weight of 10 kDa or less, and the dialysis product was freeze-dried.

Accordingly, by extracting the active ingredient from the jellyfish jelly, the membrane potential balance on the outer membrane of the mitochondria can be maintained, the activity of the mitochondria can be enhanced, and the gene expression associated with cell survival can be promoted, thereby promoting cell proliferation. And can be applied to the preparation of a system that promotes skin cell renewal The agent is the efficacy of the present invention.

Figure 1a: The extract of the post-mortem jellyfish enhances the cell viability of the keratinocyte strain HaCat.

Figure 1b: The cell survival rate of the fertility of the fibroblast cell line HS68 after the extract of the jellyfish jellyfish.

Figures 2a~2f: The extract of the jellyfish jelly promotes the number of cells in the keratinocyte strain HaCat.

Figures 3a to 3f: The extract of the jellyfish jellyfish promotes the number of cells of the fibroblast cell line HS68.

Figure 4: Jellyfish jelly extract accelerates cell healing of the keratinocyte cell line HaCat.

Figure 5a: Physiological transformation of the extract of the jellyfish jelly in the keratinocyte strain HaCat.

Figure 5b: Physiological transformation of the extract of the jellyfish jellyfish in the fibroblast cell line HS68.

The above and other objects, features and advantages of the present invention will become more <RTIgt;

The "preparation" as used in the present invention means that an active ingredient is processed and prepared into a specific dosage form, and contains a certain amount of medicines and cosmetics, and the "promoting skin cell regeneration preparation" of the present invention can maintain the granular line. The membrane potential balance on the extracorporeal membrane enhances the activity of the mitochondria and promotes gene expression associated with cell survival, which is understood by those of ordinary skill in the art to which the present invention pertains, and will not be further described herein.

The extract of the jellyfish jelly of the present invention may refer to a product obtained by concentrating the total protein of the jellyfish jellyfish. In detail, the "Queen Jellyfish" is the Cassiopea and Romeda of the Cassioidae family, and its body and wrist have symbiotic algae ( Hymenoptera ), which can be photosynthetic Role, as the most important source of nutrition for the queen jellyfish.

In the present embodiment, the extract of the queenfish jellyfish is obtained by a method comprising the steps of: taking a complete post-mortem jellyfish (including symbiotic algae) as a sample of a jellyfish jellyfish, and taking 100 grams of the jellyfish jellyfish. Sample mixed with 100 ml of phosphate buffered saline (phosphate Buffer saline (referred to as PBS), after homogenization by pre-cooled juice machine, centrifuge to obtain a supernatant (rotation speed of 6,000 ~ 10,000 rpm); continue to add ammonium sulfate solution to the supernatant (saturation percentage is 60~100%), the total protein contained in the Jellyfish sample is formed into a precipitate, and centrifuged to obtain the precipitate (rpm 6,000-10,000 rpm); then, using 50 mM Tris-HCl buffer back Dissolving the precipitate; finally dialysis using 5 liters of Tris-HCl buffer at low temperature to remove small molecular substances having a molecular weight of 10 kDa or less, and avoiding the salt in the precipitate from interfering with subsequent experiments, and The dialysis product is freeze-dried to obtain the extract of the jellyfish. In order to maintain the activity of the extract of the jellyfish jelly, each step of the above preparation method was carried out at 4 °C.

The extract of the jellyfish of the present invention can maintain the membrane potential balance on the outer membrane of the mitochondria, enhance the activity of the mitochondria, and promote the gene expression related to the survival of the cells, thereby promoting cell proliferation and can be applied to Preparing a preparation for promoting skin cell regeneration, the post-mortem jelly extract may be combined with at least one pharmaceutically acceptable carrier to form the preparation, or an additional active ingredient of the whitening agent may be added to the preparation for co-administering one The desired individual, the carrier is an excipient or an additive, and preferably can form a preparation of various forms such as a liquid, an emulsion, a cream or a gel, thereby facilitating the use of the desired individual. .

In order to confirm that the extract of the jellyfish jelly of the present invention does have an activity of promoting skin cell regeneration, the keratinocyte strain HaCat and the fibroblast strain HS68 were used as model cell strains, and the following tests were carried out:

(A) Effect on cell viability of skin cells:

Referring to the first table, the extract of the queenfish jellyfish is dissolved in the phosphate buffered saline, and then added to the model cell strain. After 24 hours of culture, the cell strains of each group are calculated by MTT assay. Cell viability, the results are shown in Figures 1a and 1b, respectively.

Table 1 and test conditions for each group of the test.

Referring to Figures 1a and 1b, the addition of the extract of the queenfish jelly can promote cell survival, whether it is HaCat cells or HS68 cells.

(B) Effects on cell proliferation of skin cells:

Continued, as shown in Table 2, the extract of the queenfish jellyfish was added to the above-mentioned model cell strain, and after 24 hours of culture, the number of cells in the visual field was observed by a microscope, and the results are shown in Fig. 2a to 3f, respectively.

Please refer to Figures 2a~2f for the addition of the extract of the Jellyfish to promote the proliferation of HaCat cells, and the same results can be observed in HS68 cells as shown in Figures 3a-3f.

(C) Effects on wound healing of skin cells:

Please also refer to the third table, the extract of the jellyfish jelly was added to HaCat cells, and after 12, 24, 36, and 48 hours of culture, the effect of the extract of the jellyfish jelly on wound healing was observed. Figure 4 shows.

Referring to Fig. 4, the addition of the extract of the jellyfish jelly can promote wound healing of HaCat cells, and the promoting effect thereof is obviously dose-dependent.

(D) Mechanism of action of the extract of the jellyfish jellyfish:

In addition, please refer to the fourth table, add the extract of the jellyfish jelly to the above-mentioned model cell strain, and after 24 hours of culture, collect the cell lysate of each group of cells, and detect and mitochondria by Western blotting method. The protein expression of the target protein related to activity and cell survival is shown in Figures 5a and 5b, respectively.

Please refer to Figure 5a for the extraction of the jellyfish jelly in HaCat cells. Adding Bcl-2, Mcl-1, Bcl-xL, Ki-67, Survivin, STAT and other proteins that inhibit the apoptotic pathway, and reducing the protein such as Bad and Bax that promote the apoptotic pathway The performance of the extract of the jellyfish jelly can also promote the protein expression of telomerase, indicating that the extract of the jellyfish jelly can maintain the membrane potential balance on the outer membrane of the mitochondria and enhance the activity of the mitochondria. And promote cell survival. As shown in Figure 5b, the same results were observed in HS68 cells.

In summary, by extracting the active ingredient from the jellyfish jelly, the membrane potential balance on the outer membrane of the mitochondria can be maintained, the activity of the mitochondria can be enhanced, and the gene expression associated with cell survival can be promoted, thereby promoting cell proliferation. It can also be applied to the preparation of a preparation for promoting skin cell renewal, which is an effect of the present invention.

While the invention has been described in connection with the preferred embodiments described above, it is not intended to limit the scope of the invention. The technical scope of the invention is protected, and therefore the scope of the invention is defined by the scope of the appended claims.

Claims (1)

The use of a jellyfish jelly extract for preparing a preparation for promoting skin cell regeneration, wherein the extract of the centipede jellyfish is obtained by a method comprising the steps of: providing a sample of a jellyfish jelly; the jellyfish jellyfish After the sample is homogenized with the phosphate buffered saline, a supernatant is obtained; the total protein of the extract of the jellyfish is precipitated with an ammonium sulfate solution (saturated percentage of 60-100%), and a precipitate is obtained; The precipitate was dissolved in 50 mM Tris-HCl buffer; and dialyzed using the Tris-HCl buffer to remove small molecular substances having a molecular weight of 10 kDa or less, and the dialysis product was freeze-dried.