{"id":4903,"date":"2024-01-15T06:33:58","date_gmt":"2024-01-15T06:33:58","guid":{"rendered":"https:\/\/corp.farlong.com\/protective-effect-of-tisochrysis-lutea-on-dry-eye-syndrome-via-nf-%ce%bab-inhibition\/"},"modified":"2024-01-27T10:30:56","modified_gmt":"2024-01-27T10:30:56","slug":"protective-effect-of-tisochrysis-lutea-on-dry-eye-syndrome-via-nf-%ce%bab-inhibition","status":"publish","type":"post","link":"https:\/\/directcm.com\/ko\/protective-effect-of-tisochrysis-lutea-on-dry-eye-syndrome-via-nf-%ce%bab-inhibition\/","title":{"rendered":"NF-\u03baB \uc5b5\uc81c\ub97c \ud1b5\ud55c \uc548\uad6c\uac74\uc870\uc99d\uc5d0 \ub300\ud55c Tisochrysis lutea\uc758 \ubcf4\ud638 \ud6a8\uacfc"},"content":{"rendered":"
\n

\uc7ac\ub8cc<\/h3>\n

\uc0dd\uccb4\ub0b4\/\uccb4\uc678 \uc7ac\ub8cc<\/h4>\n

T. \ub8e8\ud14c\uc544<\/i> Microalgae Ask Us Corp. (\uac15\ub989, \ud55c\uad6d)\uc5d0\uc11c \uc785\uc218\ud588\uc2b5\ub2c8\ub2e4. \uc2a4\ucf54\ud3f4\ub77c\ubbfc, \ud50c\ub8e8\uc624\ub808\uc138\uc778, \ud5e4\ub9c8\ud1a1\uc2e4\ub9b0 \ubc0f \uc5d0\uc624\uc2e0(H&E)\uc740 \ub3d9\ubb3c \uc2e4\ud5d8\uc744 \uc704\ud574 Sigma-Aldrich(\ubbf8\uad6d \ubbf8\uc8fc\ub9ac\uc8fc \uc138\uc778\ud2b8\ub8e8\uc774\uc2a4)\uc5d0\uc11c \uad6c\uc785\ud588\uc2b5\ub2c8\ub2e4. Dulbecco's Modified Eagle's Medium(DMEM):F12, Dulbecco \uc778\uc0b0\uc5fc \uc644\ucda9 \uc2dd\uc5fc\uc218(DPBS), \ud0dc\uc544 \uc18c \ud608\uccad(FBS) \ubc0f 0.25% \ud2b8\ub9bd\uc2e0-EDTA \uc6a9\uc561\uc740 Life Technologies(Carlsbad, CA, USA)\uc5d0\uc11c \uad6c\uc785\ud588\uc2b5\ub2c8\ub2e4. 10,000 U\/mL \ud398\ub2c8\uc2e4\ub9b0\uacfc 10,000 \u03bcg\/mL \uc2a4\ud2b8\ub819\ud1a0\ub9c8\uc774\uc2e0\uc73c\ub85c \uad6c\uc131\ub41c \ud56d\uc0dd\uc81c \uc6a9\uc561\uc740 Hyclone Laboratories Inc.(South Logan, UT, USA)\uc5d0\uc11c \uad6c\uc785\ud588\uc2b5\ub2c8\ub2e4. \uc7ac\uc870\ud569 \uc778\uac04 TNF-\u03b1\ub294 R&D \uc2dc\uc2a4\ud15c(Minneapolis, MN, USA)\uc5d0\uc11c \uc785\uc218\ud588\uc2b5\ub2c8\ub2e4. \ud56d\uc138\ud3ec\uc678 \uc2e0\ud638 \uc870\uc808 \ud0a4\ub098\uc544\uc81c(ERK), \ud56d-c-jun N-\ub9d0\ub2e8 \ud0a4\ub098\uc544\uc81c(JNK), \ud56d-p38, \ud56d-\u03b1-\ud29c\ubd88\ub9b0 \ubc0f \ud56d-\u03b2-\uc561\ud2f4\uc744 \ud3ec\ud568\ud55c 1\ucc28 \ud56d\uccb4\ub294 Santa Cruz Biotechnology\uc5d0\uc11c \uad6c\uc785\ud588\uc2b5\ub2c8\ub2e4. \ubbf8\uad6d \uce98\ub9ac\ud3ec\ub2c8\uc544\uc8fc \uc0b0\ud0c0\ud06c\ub8e8\uc988). \ub2e4\ub978 1\ucc28 \ud56d\uccb4\uc640 HRP(horseradish peroxydase) \uacb0\ud569 2\ucc28 \ud56d\uccb4\ub294 Cell Signaling Technology, Inc(Beverly, MA, USA)\uc5d0\uc11c \uad6c\uc785\ud588\uc2b5\ub2c8\ub2e4. 3-4,5-\ub514\uba54\ud2f8\ud2f0\uc544\uc878-2-\uc77c)-2,5-\ub514\ud398\ub2d0\ud14c\ud2b8\ub77c\uc878\ub968 \ube0c\ub85c\ub9c8\uc774\ub4dc(MTT)\ub97c \ud3ec\ud568\ud55c \ud654\ud559 \ubb3c\uc9c8\uc740 Sigma-Aldrich(\ubbf8\uad6d \ubbf8\uc8fc\ub9ac\uc8fc \uc138\uc778\ud2b8\ub8e8\uc774\uc2a4)\uc5d0\uc11c \uad6c\uc785\ud588\uc2b5\ub2c8\ub2e4.<\/p>\n

\ud654\ud559 \ubd84\uc11d \uc790\ub8cc<\/h3>\n

PUFA\ub294 Sigma-Aldrich(St. Louis, MO, USA)\uc5d0\uc11c \uad6c\uc785\ud55c 37\uac00\uc9c0 PUFA \ud45c\uc900 \ubb3c\uc9c8\uc774 \ud3ec\ud568\ub41c \uc6a9\uc561\uc744 \uc0ac\uc6a9\ud558\uc5ec \ud655\uc778\ub418\uc5c8\uc2b5\ub2c8\ub2e4. \ubc1c\ub808\ub974\uc0b0\uc740 \ub0b4\ubd80 \ud45c\uc900\ubb3c\uc9c8\ub85c \uc0ac\uc6a9\ud558\uae30 \uc704\ud574 Sigma-Aldrich(\ubbf8\uad6d \ubbf8\uc8fc\ub9ac\uc8fc \uc138\uc778\ud2b8\ub8e8\uc774\uc2a4)\uc5d0\uc11c \uad6c\uc785\ud588\uc2b5\ub2c8\ub2e4. \uc0bc\ubd88\ud654\ubd95\uc18c \uba54\ud0c4\uc62c \uc6a9\uc561\uc744 \ud3ec\ud568\ud55c \uc6a9\ub9e4, N<\/i>-\ud5e5\uc0b0, Na2<\/sub>\uadf8\ub798\uc11c4<\/sub>, \ucc44\ub1103<\/sub>PUFA\ub97c \ucd94\ucd9c\ud558\ub294 \ub370 \uc0ac\uc6a9\ub418\ub294 Cl, MeOH \ubc0f NaCl T. \ub8e8\ud14c\uc544<\/i> Sigma-Aldrich (\ubbf8\uad6d \ubbf8\uc8fc\ub9ac \uc8fc \uc138\uc778\ud2b8\ub8e8\uc774\uc2a4)\uc5d0\uc11c \uad6c\uc785\ud588\uc2b5\ub2c8\ub2e4.<\/p>\n

\uc724\ub9ac\uc801 \uc131\uba85<\/h3>\n

\ubcf8 \uc5f0\uad6c\ub294 \ud55c\uad6d\uacfc\ud559\uae30\uc220\uc5f0\uad6c\uc6d0(KIST) \ub3d9\ubb3c\uc2e4\ud5d8\uc5f0\uad6c\uc704\uc6d0\ud68c(IACUC)\uc758 \uc2b9\uc778\uc744 \ubc1b\uc558\uc2b5\ub2c8\ub2e4: KIST \uc81c2020-002\ud638, \uac15\ub989\uc5f0\uad6c\uc18c. \uc548\uacfc \ubc0f \uc2dc\ub825 \uc5f0\uad6c\uc5d0 \ub3d9\ubb3c\uc744 \uc0ac\uc6a9\ud569\ub2c8\ub2e4. \ub610\ud55c, \uc6b0\ub9ac\ub294 \ub3d9\ubb3c \uc5f0\uad6c: \uc0dd\uccb4 \ub0b4 \uc2e4\ud5d8 \ubcf4\uace0(ARRIVE) \uad8c\uc7a5 \uc0ac\ud56d\uc744 \ub530\ub985\ub2c8\ub2e4. \ubaa8\ub4e0 \uc808\ucc28\ub294 \ud574\ub2f9 \uaddc\uc815 \ubc0f \uc9c0\uce68\uc744 \uc900\uc218\ud558\uc5ec \uc218\ud589\ub418\uc5c8\uc2b5\ub2c8\ub2e4.<\/p>\n

\ub3d9\ubb3c \uc2e4\ud5d8 \ubc0f DES \ubaa8\ub378 \uc720\ub3c4<\/h3>\n

\ub9c8\uc6b0\uc2a4\ub294 \uc5d0\uc5b4\ucee8\uc774 \uc124\uce58\ub41c \ub3d9\ubb3c\uc2e4\uc5d0 \ubcf4\uad00\ub418\uc5c8\uc73c\uba70 22\u00b0C \u00b1 2\u00b0C, 50% \u00b1 10% \uc2b5\ub3c4 \ubc0f 12\uc2dc\uac04 \ube5b\/12\uc2dc\uac04 \uc5b4\ub450\uc6b4 \uc77c\uc8fc\uae30 \uc8fc\uae30\ub85c \uc720\uc9c0\ub418\uc5c8\uc2b5\ub2c8\ub2e4. \uc74c\uc2dd\uacfc \ubb3c\uc740 \uc790\uc720\ub86d\uac8c \uc81c\uacf5\ub418\uc5c8\uc2b5\ub2c8\ub2e4. \ub9c8\uc6b0\uc2a4\ub97c \uc77c\uc8fc\uc77c \ub3d9\uc548 \uc801\uc751\uc2dc\ud0a8 \ud6c4 \ubb34\uc791\uc704\ub85c 6\uc8fc\ub839 \uc218\ucef7 BALB\/c \ub9c8\uc6b0\uc2a4(\uc624\ub9ac\uc5d4\ud2b8\ubc14\uc774\uc624, \uacbd\uae30\ub3c4) 7\ub9c8\ub9ac\ub97c 5\uac1c \uadf8\ub8f9\uc73c\ub85c \ub098\ub204\uc5c8\ub2e4. DES\ub294 \uc778\uc0b0\uc5fc \uc644\ucda9 \uc2dd\uc5fc\uc218(PBS; Sigma-Aldrich, St. Louis, MO, USA)\uc5d0 \ud76c\uc11d\ub41c \uc2a4\ucf54\ud3f4\ub77c\ubbfc 200 \u03bcL(2.5 mg\/mL)\ub97c \ubcf5\uac15 \ub0b4 \uc8fc\uc0ac\ud558\uc5ec \ub9e4\uc77c 2\ud68c \ub9c8\uc6b0\uc2a4\uc5d0\uc11c \uc2e4\ud5d8\uc801\uc73c\ub85c \uc720\ub3c4\ub418\uc5c8\uc2b5\ub2c8\ub2e4. T. \ub8e8\ud14c\uc544<\/i> 200 \u03bcL\uc758 \uc99d\ub958\uc218\uc5d0 0(\ucc28\ub7c9 \ub300\uc870\uad70), 100, 150 \ub610\ub294 300 mg\/kg\uc758 \ub18d\ub3c4\ub85c \ub9e4\uc77c \ub9c8\uc6b0\uc2a4 \uadf8\ub8f9\uc5d0 \uacbd\uad6c \ud22c\uc5ec\ud588\uc2b5\ub2c8\ub2e4. \ub300\uc870\uad70\uc5d0\ub294 \uc2a4\ucf54\ud3f4\ub77c\ubbfc\uc774 \ud3ec\ud568\ub418\uc9c0 \uc54a\uc740 PBS\ub97c \ud22c\uc5ec\ud588\uc2b5\ub2c8\ub2e4. \uc5b8\uc81c T. \ub8e8\ud14c\uc544<\/i> \uacbd\uad6c\ud22c\uc5ec\ud558\uc600\uace0 \ub300\uc870\uad70\uacfc DES\uad70\uc5d0\uc11c\ub294 \uc99d\ub958\uc218 200 \u03bcL\ub97c \uc0ac\uc6a9\ud558\uc600\ub2e4. \ub208\ubb3c \uc0dd\uc0b0\uc740 1\ubd84 \ub3d9\uc548 \ub208\uc744 \uac10\uae30 \uc804 \uad00\uc790\ub180\uc774 \ub208\uaebc\ud480\uc758 \uc544\ub798\ucabd 1\/3\uc5d0 \uc704\uce58\ud55c \ud45c\uc900 Schirmer \ud14c\uc2a4\ud2b8 \uc2a4\ud2b8\ub9bd\uc744 \uc0ac\uc6a9\ud558\uc5ec 2\uc8fc \ud6c4\uc5d0 \uc815\ub7c9\ud654\ub418\uc5c8\uc2b5\ub2c8\ub2e4. \uadf8\ub7f0 \ub2e4\uc74c \uc2a4\ud2b8\ub9bd\uc744 \uc81c\uac70\ud558\uace0 \ubc00\ub9ac\ubbf8\ud130 \ub2e8\uc704\uc758 \uc2b5\uc724\uc810 \uae38\uc774\ub97c \uce21\uc815\ud558\uc5ec Schirmer\uc758 \ud14c\uc2a4\ud2b8 \uac12\uc744 \uacb0\uc815\ud588\uc2b5\ub2c8\ub2e4. \uac01\ub9c9 \ud45c\uba74\uc758 \uc5fc\uc0c9\uc740 \uac01\ub9c9 \ud45c\uba74 \ubcc0\ud654\uc758 \uc815\ub3c4\ub97c \uacb0\uc815\ud558\ub294 \ub370 \uc0ac\uc6a9\ub418\uc5c8\uc2b5\ub2c8\ub2e4. 3% \ud50c\ub8e8\uc624\ub808\uc138\uc778\uc744 \ud558\uce21 \uacb0\ub9c9\uc5d0 \ud55c \ubc29\uc6b8 \uc8fc\uc785\ud55c \ud6c4 \uac01\ub9c9 \ud45c\uba74\uc744 \uad00\ucc30\ud558\uace0 \uc810\uc218\ub97c \ub9e4\uacbc\ub2e4. \uac01\ub9c9 \uc5fc\uc0c9\uc740 \ub9f9\ubaa9\uc801\uc73c\ub85c \ud3c9\uac00\ub418\uc5c8\uc2b5\ub2c8\ub2e4. \ub9c8\uc6b0\uc2a4\ub294 \uacbd\ucd94 \ud0c8\uad6c\ub85c \uc778\ud574 \uc548\ub77d\uc0ac\ub418\uc5c8\uc2b5\ub2c8\ub2e4. \uac01\ub9c9\uc740 \ub9c8\uc6b0\uc2a4\uc758 \ub208\uaebc\ud480\uc744 \uac00\ubccd\uac8c \ub450\ub4dc\ub9b0 \ud6c4 \uc9d1\uac8c\ub97c \uc0ac\uc6a9\ud558\uc5ec \uc548\uad6c\uc5d0\uc11c \uc2dc\uc2e0\uacbd\uc744 \uc798\ub77c\ub0b4\uc5b4 \uac01\ub9c9\uc744 \uc81c\uac70\ud588\uc2b5\ub2c8\ub2e4. \uc218\uc815\uccb4\ub294 \uac01\ub9c9 \ub0b4\uc5d0\uc11c \uc81c\uac70\ub418\uc5b4 -80\u00b0C\uc5d0 \ubcf4\uad00\ub418\uc5c8\uc2b5\ub2c8\ub2e4. \uc9d1\uac8c\ub97c \uc0ac\uc6a9\ud558\uc5ec \ub9c8\uc6b0\uc2a4\uc758 \uc544\ub798\ud131\uc5d0\uc11c \ub208\ubb3c\uc0d8\uc744 \uc81c\uac70\ud558\uace0, \ub2f9\uae34 \ubd80\ubd84\uc758 \ud45c\ud53c\ub97c \uc798\ub77c\ub0b4\uc5b4 \uc5bc\uad74\uc758 \uc544\ub798\ucabd \uc808\ubc18\uc744 \ub4dc\ub7ec\ub0c8\uc2b5\ub2c8\ub2e4. \uc544\ub798\ud131 \uadfc\ucc98\uc758 \ub208\ubb3c\uc0d8\uc744 \uace0\uc815\ud558\uace0 \uc81c\uac70\ud55c \ud6c4 \ube5b\uc744 \ucc28\ub2e8\ud558\uc5ec -80 \u00b0C\uc5d0 \ubcf4\uad00\ud588\uc2b5\ub2c8\ub2e4.<\/p>\n

\uc870\uc9c1\ud559<\/h3>\n

\uac01\ub9c9 \ud45c\ud53c \ubc0f \ub208\ubb3c\uc0d8 \uc870\uc9c1\uc744 \uc218\uc9d1\ud558\uc5ec 10% \ud3ec\ub984\uc54c\ub370\ud788\ub4dc\uc5d0 \uace0\uc815\ud55c \ud6c4 \ud30c\ub77c\ud540 \ud3ec\ub9e4 \ubc0f \uc808\uac1c\ub97c \uc704\ud574 \ucc98\ub9ac\ud588\uc2b5\ub2c8\ub2e4. \ub2e8\uba74\uc744 H&E\ub85c \uc5fc\uc0c9\ud558\uace0 \ud604\ubbf8\uacbd(TE-2000U, Nikon, Tokyo, Japan)\uc744 \uc0ac\uc6a9\ud558\uc5ec 40\ubc30\uc728\ub85c \uac80\uc0ac\ud588\uc2b5\ub2c8\ub2e4. \uc911\uc559 \uac01\ub9c9 \uc0c1\ud53c \ub450\uaed8\ub294 \uac01 \uac01\ub9c9\uc744 5\uac1c \ubd80\ubd84\uc73c\ub85c \ub098\ub204\uc5b4 \uce21\uc815\ud588\uc2b5\ub2c8\ub2e4.<\/p>\n

\uba74\uc5ed\uc870\uc9c1\ud654\ud559\uc801(IHC) \ubd84\uc11d\uc744 \uc704\ud55c DAB \uc5fc\uc0c9<\/h3>\n

BALB\/c \ub9c8\uc6b0\uc2a4\uc758 \ub208\ubb3c\uc0d8 \uc870\uc9c1\uc744 \ucc44\ucde8\ud558\uc5ec 10% \ud3ec\ub984\uc54c\ub370\ud788\ub4dc\ub85c \uace0\uc815\ud588\uc2b5\ub2c8\ub2e4. \uace0\uc815\ub41c \ub208\ubb3c\uc0d8 \uc870\uc9c1\uc744 \ud30c\ub77c\ud540\uc73c\ub85c \ud3ec\ub9e4\ud558\uace0 \uc808\ud3b8(5 \u03bcm)\ud55c \ud6c4 xylene(Jonsei, Tokyo, Japan)\uc744 \uc0ac\uc6a9\ud558\uc5ec 3\ubd84\uac04 3\ud68c\uc5d0 \uac78\uccd0 \ud30c\ub77c\ud540\uc744 \uc81c\uac70\ud588\uc2b5\ub2c8\ub2e4. \uc808\ud3b8\uc744 100, 95, 70 \ubc0f 50% \uc5d0\ud0c4\uc62c\uc5d0\uc11c 1\ubd84 \ub3d9\uc548 \uc218\ud654\uc2dc\ucf30\uc2b5\ub2c8\ub2e4. TRIS-EDTA \uc644\ucda9\uc561(pH 9.0)\uc744 \uc0ac\uc6a9\ud558\uc5ec \uc804\uc790\ub808\uc778\uc9c0\ub85c \ub208\ubb3c\uc0d8 \ud56d\uc6d0\uc744 \ucd94\ucd9c\ud588\uc2b5\ub2c8\ub2e4. 3% \uacfc\uc0b0\ud654\uc218\uc18c\ub97c \uc870\uc9c1 \uc2ac\ub77c\uc774\ub4dc\uc5d0 15\ubd84 \ub3d9\uc548 \uc801\uc6a9\ud55c \ud6c4, \uc808\ud3b8\uc744 \uc2e4\uc628\uc5d0\uc11c 1\uc2dc\uac04 \ub3d9\uc548 2% BSA\ub85c \ucc28\ub2e8\ud588\uc2b5\ub2c8\ub2e4. \uc808\ud3b8\uc744 CD45(Bio-RAD, MCA 1258GT, 1:100 \ud76c\uc11d)\ub85c 4\u00b0C\uc5d0\uc11c \ubc24\uc0c8 \ucc98\ub9ac\ud588\uc2b5\ub2c8\ub2e4. PBS\ub85c \uc138\ucc99\ud55c \ud6c4 \uc138\ud3ec\ub97c \uc2e4\uc628\uc5d0\uc11c 2\uc2dc\uac04 \ub3d9\uc548 \ud56d\ud1a0\ub07c IgG-HRP(Santa Cruz Biotechnology, Dallas, TX, USA, SC-2357, 1:100 \ud76c\uc11d)\ub85c \ucc98\ub9ac\ud558\uace0 3, 3'-\ub514\uc544\ubbf8\ub178\ubca4\uc9c0\ub518 \ud14c\ud2b8\ub77c\ud788\ub4dc\ub85c\ud074\ub85c\ub77c\uc774\ub4dc(Vector Laboratories, Burlingame, CA, USA). \uc2ac\ub77c\uc774\ub4dc\ub97c \ud5e4\ub9c8\ud1a1\uc2e4\ub9b0\uc73c\ub85c 30\ucd08 \ub3d9\uc548 \uc5fc\uc0c9\ud558\uace0 \uc218\ub3d7\ubb3c\ub85c \ud5f9\uad6c\uc5c8\uc2b5\ub2c8\ub2e4. \uadf8\ub7f0 \ub2e4\uc74c \uc2ac\ub77c\uc774\uc2a4\ub97c \ub36e\uac1c\ub85c \ub36e\uace0 Permount\u2122 \uc7a5\ucc29 \ub9e4\uccb4(Thermo Fisher Scientific, Waltham, MA, USA)\ub97c \uc0ac\uc6a9\ud558\uc5ec \uc7a5\ucc29\ud588\uc2b5\ub2c8\ub2e4. \ubaa8\ub4e0 \uc2ac\ub77c\uc774\ub4dc\ub294 \uad11\ud559 \ud604\ubbf8\uacbd(Olympus, CX43, Olympus Optical Co., Tokyo, Japan)\uc744 \uc0ac\uc6a9\ud558\uc5ec \uac80\uc0ac\ud558\uace0 \uc0ac\uc9c4\uc744 \ucc0d\uc5c8\uc2b5\ub2c8\ub2e4. ImageJ \uc18c\ud504\ud2b8\uc6e8\uc5b4(\ubc84\uc804 1.53, \ubbf8\uad6d \uba54\ub9b4\ub79c\ub4dc\uc8fc \ubca0\ub370\uc2a4\ub2e4 \uad6d\ub9bd\ubcf4\uac74\uc6d0)48<\/a><\/sup> \uc815\ub7c9\uc801 \ub370\uc774\ud130 \ubd84\uc11d\uc744 \uc218\ud589\ud558\ub294 \ub370 \uc0ac\uc6a9\ub418\uc5c8\uc2b5\ub2c8\ub2e4.<\/p>\n

ARPE-19 \uc138\ud3ec \ubc30\uc591<\/h3>\n

\uc778\uac04 \ub9dd\ub9c9 \uc0c1\ud53c ARPE-19 \uc138\ud3ec(American Type Culture Collection, ATCC, Manassas, VA, USA)\ub97c 10% FBS(HyClone Laboratories, Logan, UT)\uac00 \ubcf4\ucda9\ub41c DMEM\/F-12 \ubc30\uc9c0(Gibco, Carlsbad, CA, USA)\uc5d0\uc11c \ubc30\uc591\ud588\uc2b5\ub2c8\ub2e4. , USA) \ubc0f 1% \ud398\ub2c8\uc2e4\ub9b0\/\uc2a4\ud2b8\ub819\ud1a0\ub9c8\uc774\uc2e0(HyClone Laboratories). \uc138\ud3ec\ub294 2 \u00d7 10 \uc758 \ubc00\ub3c4\ub85c \ud30c\uc885\ub418\uc5c8\uc2b5\ub2c8\ub2e4.5<\/sup> 6\uc6f0 \ud50c\ub808\uc774\ud2b8\uc5d0 \uc6f0\ub2f9 \ubd84\uc8fc\ud558\uace0 \ubc30\uc591\ud558\uc5ec \uc5fc\uc99d \ubc0f \uc18c\ud3ec\uccb4 \ubc18\uc751\uc744 \uc720\ub3c4\ud588\uc2b5\ub2c8\ub2e4.<\/p>\n

\uc138\ud3ec \uc0dd\uc874\ub825<\/h3>\n

\ud6a8\uacfc T. \ub8e8\ud14c\uc544<\/i> \uc138\ud3ec \uc0dd\uc874\ub825\uc5d0 \ub300\ud55c \uc774\uc804 \uc5f0\uad6c\uc5d0\uc11c \uc124\uba85\ud55c \ub300\ub85c MTT \ubd84\uc11d\uc73c\ub85c \ud3c9\uac00\ud588\uc2b5\ub2c8\ub2e4.49<\/a><\/sup> \ud558\uc9c0\ub9cc \uc57d\uac04 \uc218\uc815\ud588\uc2b5\ub2c8\ub2e4. \uc138\ud3ec\ub97c 96\uc6f0 \ubc30\uc591 \ud50c\ub808\uc774\ud2b8\uc5d0 1.5 x 10 \ubc00\ub3c4\ub85c \ud50c\ub808\uc774\ud305\ud588\uc2b5\ub2c8\ub2e4.4<\/sup> \uc138\ud3ec\/\uc6f0\uc744 \ucc98\ub9ac\ud558\uace0 24\uc2dc\uac04 \ub3d9\uc548 \ubc30\uc591\ud588\uc2b5\ub2c8\ub2e4. \uadf8\ub7f0 \ub2e4\uc74c \uc138\ud3ec\ub97c \ub2e4\uc591\ud55c \ub18d\ub3c4\uc758 T. \ub8e8\ud14c\uc544<\/i> \uadf8\ub9ac\uace0 \ucd94\uac00\ub85c 24\uc2dc\uac04 \ub3d9\uc548 \ubc30\uc591\ud558\uc600\ub2e4. \uac01 \uc6f0\uc758 \uc0c1\uccad\uc561\uc744 0.5 mg\/mL\uc758 MTT\ub97c \ud568\uc720\ud55c \uc0c8\ub85c\uc6b4 \uc131\uc7a5 \ubc30\uc9c0\ub85c \uad50\uccb4\ud558\uace0 1\uc2dc\uac04 \ub3d9\uc548 \ubc30\uc591\ud588\uc2b5\ub2c8\ub2e4. \uc0c1\ub4f1\uc561\uc744 \uc81c\uac70\ud558\uace0 \uc0dd\uc131\ub41c \ud3ec\ub974\ub9c8\uc794 \uce68\uc804\ubb3c\uc744 200 \u03bcL\uc758 \ub514\uba54\ud2f8 \uc124\ud3ed\uc0ac\uc774\ub4dc\uc5d0 \uc6a9\ud574\uc2dc\ucf30\uc2b5\ub2c8\ub2e4. \ud761\uad11\ub3c4\ub294 570 nm\uc5d0\uc11c \uce21\uc815\ub418\uc5c8\uc2b5\ub2c8\ub2e4. \uc138\ud3ec \uc0dd\uc874\uc728\uc740 \uc74c\uc131 \ub300\uc870\uad70\uacfc \ube44\uad50\ud558\uc5ec \ubc31\ubd84\uc728\ub85c \uc0c1\ub300\uc801\uc73c\ub85c \uacc4\uc0b0\ub418\uc5c8\uc2b5\ub2c8\ub2e4.<\/p>\n

\uc6e8\uc2a4\ud134 \ube14\ub86f \ubd84\uc11d<\/h3>\n

\ud56d\uc5fc\uc99d \ud6a8\uacfc\uc758 \uae30\ubcf8\uc774 \ub418\ub294 \uc138\ud3ec \uc2e0\ud638 \uc804\ub2ec \uacbd\ub85c\ub97c \uacb0\uc815\ud558\uae30 \uc704\ud574 \uc6e8\uc2a4\ud134 \ube14\ub86f\ud305\uc744 \uc218\ud589\ud588\uc2b5\ub2c8\ub2e4. T. \ub8e8\ud14c\uc544<\/i> \uc55e\uc11c \uc124\uba85\ud55c \ub300\ub85c \uc5f0\uad6c49<\/a><\/sup>, \uadf8\ub7ec\ub098 \uc57d\uac04\uc758 \uc218\uc815\uc774 \ud544\uc694\ud569\ub2c8\ub2e4. \uc735\ud569\uc131 ARPE-19 \uc138\ud3ec\ub294 1% FBS \ubc0f 1% \ud56d\uc0dd\uc81c\ub97c \ud568\uc720\ud55c DMEM:F12\uc5d0\uc11c 24\uc2dc\uac04 \ub3d9\uc548 \uad76\uc5b4 \uc8fd\uc5c8\uc2b5\ub2c8\ub2e4. \uadf8\ub7f0 \ub2e4\uc74c \uc138\ud3ec\ub97c \ub2e4\uc74c\uc73c\ub85c \ucc98\ub9ac\ud588\uc2b5\ub2c8\ub2e4. T. \ub8e8\ud14c\uc544<\/i> (50 \ubc0f 100 \u03bcg\/mL)\uc744 2\uc2dc\uac04 \ub3d9\uc548 \ucc98\ub9ac\ud55c \ud6c4 20 ng\/mL\uc758 TNF-\u03b1\ub85c 15\ubd84 \ub3d9\uc548 \uc790\uadf9\ud588\uc2b5\ub2c8\ub2e4. \uc138\ud3ec\ub97c \uc5bc\uc74c\ucc98\ub7fc \ucc28\uac00\uc6b4 DPBS\ub85c 3\ud68c \ud5f9\uad6c\uace0 \ud504\ub85c\ud14c\uc544\uc81c\uc640 \ud3ec\uc2a4\ud30c\ud0c0\uc81c \uc5b5\uc81c\uc81c\uc758 \uce75\ud14c\uc77c\uc774 \ubcf4\ucda9\ub41c \ub2e8\ubc31\uc9c8 \ucd94\ucd9c \uc644\ucda9\uc561(iNtRON Biotechnology, \uacbd\uae30\ub3c4, \ud55c\uad6d)\uc5d0 \uc6a9\ud574\uc2dc\ucf30\ub2e4. \uc6a9\ud574\ubb3c\uc758 \uc0c1\uccad\uc561\uc744 \uc6d0\uc2ec\ubd84\ub9ac(14,000\u00d7g<\/i>, 30\ubd84, 4\u00b0C), DCTM \ub2e8\ubc31\uc9c8 \ubd84\uc11d \ud0a4\ud2b8(Bio-Rad, Hercules, CA, USA)\ub97c \uc0ac\uc6a9\ud558\uc5ec \ub2e8\ubc31\uc9c8 \ub18d\ub3c4\ub97c \ucd94\uc815\ud588\uc2b5\ub2c8\ub2e4. \ub3d9\uc77c\ud55c \uc591(15-25\u03bcg)\uc758 \uc138\ud3ec \ub2e8\ubc31\uc9c8\uc744 SDS-PAGE\ub85c \ubd84\ub9ac\ud558\uace0 PVDF \ub9c9\uc73c\ub85c \uc62e\uacbc\uc2b5\ub2c8\ub2e4. \ub9c9\uc744 5% \uc18c \ud608\uccad \uc54c\ubd80\ubbfc \ub610\ub294 \ud0c8\uc9c0\uc720\uc640 \ud568\uaed8 \ubc30\uc591\ud55c \ub2e4\uc74c 1\ucc28 \ud56d\uccb4(\ud56d-\ud3ec\uc2a4\ud3ec-ERK, \ud56d-ERK, \ud56d-\ud3ec\uc2a4\ud3ec-JNK, \ud56d-JNK, \ud56d-\ud3ec\uc2a4\ud3ec-p38)\uc758 1:2000 \ud76c\uc11d\uacfc \ud568\uaed8 \ubc30\uc591\ud588\uc2b5\ub2c8\ub2e4. , \ud56d-p38, \ud56d-\ud3ec\uc2a4\ud3ec-AKT, \ud56d-AKT, \ud56d-\ud3ec\uc2a4\ud3ec-I\u03baB-\u03b1, \ud56d-I\u03baB-\u03b1, \ud56d-\ud3ec\uc2a4\ud3ec-p65, \ud56d-p65, \ud56d-\u03b1-\ud29c\ubd88\ub9b0 \ubc0f \ud56d-\u03b2-\uc791\uc6a9 ) 4\u00b0C \ubc0f 12\uc2dc\uac04. \uadf8\ub7f0 \ub2e4\uc74c \ub9c9\uc744 TBS-T\ub85c \ud5f9\uad6c\uace0 1:10,000\uc73c\ub85c \ud76c\uc11d\ub41c \ud56d\ud1a0\ub07c HRP \uacb0\ud569 2\ucc28 \ud56d\uccb4\ub85c \uc2e4\uc628\uc5d0\uc11c 1\uc2dc\uac04 \ub3d9\uc548 \ucc98\ub9ac\ud588\uc2b5\ub2c8\ub2e4. SuperSignal West Femto Maximum Sensitivity Substrate(Thermo Fisher Scientific)\ub97c \uc0ac\uc6a9\ud558\uc5ec \ub2e8\ubc31\uc9c8 \ubc34\ub4dc\ub97c \uac80\ucd9c\ud558\uace0 iBright CL1000 \uac94 \ubb38\uc11c\ud654 \uc2dc\uc2a4\ud15c(Thermo Fisher Scientific)\uc5d0\uc11c \uc2dc\uac01\ud654\ud588\uc2b5\ub2c8\ub2e4. \uac01 \ub2e8\ubc31\uc9c8 \ube14\ub86f\uc758 \ubc00\ub3c4\ub294 Image J \uc18c\ud504\ud2b8\uc6e8\uc5b4(\ubc84\uc804 1.53, NIH)\ub97c \uc0ac\uc6a9\ud558\uc5ec \uacb0\uc815\ub418\uc5c8\uc2b5\ub2c8\ub2e4.48<\/a><\/sup>.<\/p>\n

\ucd1d PUFA \ucd94\ucd9c T. \ub8e8\ud14c\uc544<\/i><\/h3>\n

\ucd1d PUFA\ub294 \ub2e4\uc74c\uc5d0\uc11c \ucd94\ucd9c\ub418\uc5c8\uc2b5\ub2c8\ub2e4. T. \ub8e8\ud14c\uc544<\/i> \ud3f4\uce58(Folch) \uae30\ubc95\uc744 \uc0ac\uc6a9\ud558\uc5ec50<\/a><\/sup>. \uac04\ub2e8\ud788 \ub9d0\ud558\uba74, 100mg\uc758 \uc0d8\ud50c\uc744 5mL\uc758 CH\uac00 \ub4e4\uc5b4 \uc788\ub294 \ubc14\uc774\uc54c\uc5d0 \ub123\uc5c8\uc2b5\ub2c8\ub2e4.3<\/sub>Cl:MeOH(2:1, v\/v) \uc6a9\uc561\uc744 \ub123\uace0 15\ubd84 \ub3d9\uc548 \ucd08\uc74c\ud30c \ucc98\ub9ac\ud588\uc2b5\ub2c8\ub2e4. 10,000rpm\uc5d0\uc11c 5\ubd84 \ub3d9\uc548 \uc6d0\uc2ec\ubd84\ub9ac\ud558\uc5ec \ud604\ud0c1\uc561\uc744 \uc815\ud654\ud588\uc2b5\ub2c8\ub2e4. \ub0982<\/sub>\uadf8\ub798\uc11c4<\/sub> \uc0c1\ub4f1\uc561\uc5d0 \ucca8\uac00\ud55c \ud6c4 \uc5ec\uacfc\ud558\uc5ec \ub0a8\uc740 \ubb3c\uc744 \uc81c\uac70\ud588\uc2b5\ub2c8\ub2e4. \uc774 \ub2e8\uacc4\ub97c \ub450 \ubc88 \ub354 \ubc18\ubcf5\ud588\uc2b5\ub2c8\ub2e4. \uc774\uc5b4\uc11c, \uc6a9\uc561\uc744 \uc9c8\uc18c \uac00\uc2a4\ud558\uc5d0 \ub18d\ucd95\ud558\uc5ec \uc21c\uc218\ud55c \uc870\ub958 \uc9c0\uc9c8\uc744 \uc5bb\uc5c8\ub2e4.<\/p>\n

\uc9c0\ubc29\uc0b0 \uba54\ud2f8\ud654<\/h3>\n

\uac00\uc2a4 \ud06c\ub85c\ub9c8\ud1a0\uadf8\ub798\ud53c(GC) \ubd84\uc11d\uc744 \uc704\ud55c \uc9c0\uc9c8 \uc0d8\ud50c\uc744 \uc900\ube44\ud558\uae30 \uc704\ud574 \uc9c0\ubc29\uc0b0 \uba54\ud2f8\ud654\ub97c \uc218\ud589\ud588\uc2b5\ub2c8\ub2e4. \ucd94\ucd9c\ub41c \ubb3c\uc9c8\uc744 10mL \ubc14\uc774\uc54c\uc5d0 \ucca8\uac00\ud55c \ud6c4 1mL\uc758 0.5N NaOH:MeOH \ud63c\ud569\ubb3c\uc744 \ucca8\uac00\ud588\uc2b5\ub2c8\ub2e4. \uc774 \uc6a9\uc561\uc5d0\ub294 400\u03bcg\/mL\uc758 \ub0b4\ubd80 \uac00\ub2e5(\ubc1c\ub808\ub974\uc0b0)\uc774 \ud3ec\ud568\ub418\uc5b4 \uc788\uc2b5\ub2c8\ub2e4. \ud63c\ud569\ubb3c\uc744 30\ucd08 \ub3d9\uc548 \ud754\ub4e0 \ub2e4\uc74c 80\u00b0C\uc5d0\uc11c 20\ubd84 \ub3d9\uc548 \ubc30\uc591\ud588\uc2b5\ub2c8\ub2e4. \ud63c\ud569\ubb3c\uc744 \uc2e4\uc628\uc5d0\uc11c 5\ubd84 \ub3d9\uc548 \ub0c9\uac01\uc2dc\ud0a8 \ud6c4 \uc0bc\ubd88\ud654\ubd95\uc18c:MeOH 2 mL\ub97c \ucca8\uac00\ud558\uace0 30\ucd08 \ub3d9\uc548 \uad50\ubc18\ud558\uc600\ub2e4. \ud63c\ud569\ubb3c\uc744 80\u2103\uc5d0\uc11c 20\ubd84 \ub3d9\uc548 \ubc30\uc591\ud55c \ud6c4 10\ubd84 \ub3d9\uc548 \ub0c9\uac01\uc2dc\ucf30\ub2e4. 2 mL\uc758 \uacfc\ud3ec\ud654 NaCl\uc744 \ucca8\uac00\ud558\uc5ec \uc0c1 \ubd84\ub9ac \ubc0f \uc9c0\ubc29\uc0b0 \uc218\uc9d1\uc744 \uc720\ub3c4\ud588\uc2b5\ub2c8\ub2e4. \ub2e4\uc74c\uc73c\ub85c 1.5mL N<\/i>-\ud5e5\uc0b0\uc744 \ucca8\uac00\ud558\uace0 \ud63c\ud569\ubb3c\uc744 30\ucd08 \ub3d9\uc548 \uc640\ub3d9\uc2dc\ud0a8 \ud6c4 \uc2e4\uc628\uc5d0\uc11c 20\ubd84 \ub3d9\uc548 \ub0c9\uac01\uc2dc\ucf30\ub2e4. \uce35\uc744 \ubd84\ub9ac\ud558\uace0 \uc561\uccb4 \uc0c1\uce35\uc561\uc744 \ud63c\ud569\ubb3c\ub85c\ubd80\ud130 \uc81c\uac70\ud55c \ud6c4 \ud3ec\ud654 Na2\uc218\uc9c0\ub97c \uc0ac\uc6a9\ud558\uc5ec \ubb3c\uc744 \uc644\uc804\ud788 \uc81c\uac70\ud558\uc600\ub2e4.2<\/sub>\uadf8\ub798\uc11c4<\/sub> \ud544\ud130.<\/p>\n

\uc9c0\ubc29\uc0b0\uc758 \uc2dd\ubcc4<\/h3>\n

\ubd88\uaf43 \uc774\uc628\ud654 \uac80\ucd9c\uae30(FID; Agilent 7890A, Agilent Technologies, Wilmington, DE, USA)\uac00 \uc7a5\ucc29\ub41c GC\ub97c \uc0ac\uc6a9\ud558\uc5ec \uc9c0\ubc29\uc0b0 \uba54\ud2f8 \uc5d0\uc2a4\ud14c\ub974(FAME)\ub97c \uac80\uc0ac\ud588\uc2b5\ub2c8\ub2e4. \uc0d8\ud50c(1\u03bcL)\uc744 GC(J&W 112-88A7, Agilent Technologies, Wilmington, DE, USA)(100m \u00d7 250\u03bcm ID, 0.2\u03bcm \ud544\ub984)\uc758 HP-88 \uceec\ub7fc\uc5d0 \uc8fc\uc785\ud588\uc2b5\ub2c8\ub2e4. GC \ubd84\uc11d\uc740 \ub2e4\uc74c\uacfc \uac19\uc774 \uc218\ud589\ub418\uc5c8\uc2b5\ub2c8\ub2e4. \uc0d8\ud50c\uc744 \ucc98\uc74c\uc5d0 140\u00b0C\uc5d0\uc11c 5\ubd84\uac04 \uc720\uc9c0\ud55c \ub2e4\uc74c \uc628\ub3c4\ub97c \ubd84\ub2f9 4\u00b0C\uc529 \uc62c\ub824 15\ubd84 \ub3d9\uc548 240\u00b0C\uae4c\uc9c0 \uc62c\ub838\uc2b5\ub2c8\ub2e4. \uceec\ub7fc\uc758 \uc720\uc18d\uc740 1mL\/min\uc73c\ub85c \uace0\uc815\ub418\uc5c8\uc2b5\ub2c8\ub2e4. \ubd84\uc11d\uc744 \uc704\ud55c \uc6b4\ubc18\uac00\uc2a4\ub85c\ub294 \uc9c8\uc18c\uac00\uc2a4\ub97c \uc0ac\uc6a9\ud558\uc600\uc73c\uba70, FID\ub294 280\u2103, \ubd84\ud560\ube44\ub294 30:1\ub85c \uc124\uc815\ud558\uc600\ub2e4. FAME\ub294 \uc54c\ub824\uc9c4 \ud45c\uc900\uacfc \ube44\uad50\ud558\uc5ec \uc2dd\ubcc4\ub418\uc5c8\uc2b5\ub2c8\ub2e4. \uac01 \uc0d8\ud50c\uc740 GC \ubd84\uc11d\uc73c\ub85c 3\ud68c \uce21\uc815\ub418\uc5c8\uc2b5\ub2c8\ub2e4.<\/p>\n

\ud1b5\uacc4 \ubd84\uc11d<\/h3>\n

\ubaa8\ub4e0 \ub370\uc774\ud130\ub294 SPSS \ubc84\uc804 18.0(IBM Co., Armonk, NY, USA)\uc744 \uc0ac\uc6a9\ud558\uc5ec Tukey\uc758 \uc0ac\ud6c4 \ubd84\uc11d\uacfc \ud568\uaed8 \uc77c\uc6d0 \ubd84\uc0b0 \ubd84\uc11d\uc744 \uc0ac\uc6a9\ud558\uc5ec \ud1b5\uacc4\uc801\uc73c\ub85c \ubd84\uc11d\ub418\uc5c8\uc2b5\ub2c8\ub2e4. \uac01 \uc2e4\ud5d8\uc5d0 \ub300\ud574 \ucd5c\uc18c 3\ubc88\uc758 \ub3c5\ub9bd\uc801\uc778 \ubc18\ubcf5\uc2e4\ud5d8\uc774 \uc218\ud589\ub418\uc5c8\uc2b5\ub2c8\ub2e4.<\/p>\n<\/div>\n

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\uc18c\uc2a4 \ub9c1\ud06c <\/a><\/p>","protected":false},"excerpt":{"rendered":"

The prevalence of dry eye syndrome (DES) has increased worldwide, affecting around 50% of all adults1. DES is a multifactorial ocular surface disease characterized by tear film instability and hyperosmolarity, ocular surface inflammation and damage, neurosensory abnormalities<\/p>","protected":false},"author":1,"featured_media":5363,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"site-sidebar-layout":"default","site-content-layout":"","ast-site-content-layout":"default","site-content-style":"default","site-sidebar-style":"default","ast-global-header-display":"","ast-banner-title-visibility":"","ast-main-header-display":"","ast-hfb-above-header-display":"","ast-hfb-below-header-display":"","ast-hfb-mobile-header-display":"","site-post-title":"","ast-breadcrumbs-content":"","ast-featured-img":"","footer-sml-layout":"","theme-transparent-header-meta":"default","adv-header-id-meta":"","stick-header-meta":"","header-above-stick-meta":"","header-main-stick-meta":"","header-below-stick-meta":"","astra-migrate-meta-layouts":"set","ast-page-background-enabled":"default","ast-page-background-meta":{"desktop":{"background-color":"var(--ast-global-color-4)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-gradient":""},"tablet":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-gradient":""},"mobile":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-gradient":""}},"ast-content-background-meta":{"desktop":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-gradient":""},"tablet":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-gradient":""},"mobile":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-gradient":""}},"footnotes":""},"categories":[9],"tags":[],"class_list":["post-4903","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-supplements"],"_links":{"self":[{"href":"https:\/\/directcm.com\/ko\/wp-json\/wp\/v2\/posts\/4903","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/directcm.com\/ko\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/directcm.com\/ko\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/directcm.com\/ko\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/directcm.com\/ko\/wp-json\/wp\/v2\/comments?post=4903"}],"version-history":[{"count":1,"href":"https:\/\/directcm.com\/ko\/wp-json\/wp\/v2\/posts\/4903\/revisions"}],"predecessor-version":[{"id":5393,"href":"https:\/\/directcm.com\/ko\/wp-json\/wp\/v2\/posts\/4903\/revisions\/5393"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/directcm.com\/ko\/wp-json\/wp\/v2\/media\/5363"}],"wp:attachment":[{"href":"https:\/\/directcm.com\/ko\/wp-json\/wp\/v2\/media?parent=4903"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/directcm.com\/ko\/wp-json\/wp\/v2\/categories?post=4903"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/directcm.com\/ko\/wp-json\/wp\/v2\/tags?post=4903"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}