{"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\/ja\/protective-effect-of-tisochrysis-lutea-on-dry-eye-syndrome-via-nf-%ce%bab-inhibition\/","title":{"rendered":"NF-\u03baB\u963b\u5bb3\u306b\u3088\u308b\u30c9\u30e9\u30a4\u30a2\u30a4\u75c7\u5019\u7fa4\u306b\u5bfe\u3059\u308bTisochrysis lutea\u306e\u4fdd\u8b77\u52b9\u679c"},"content":{"rendered":"
\n

\u6750\u6599<\/h3>\n

\u30a4\u30f3\u30d3\u30dc\/\u30a4\u30f3\u30d3\u30c8\u30ed\u6750\u6599<\/h4>\n

T.\u30eb\u30c6\u30a2<\/i> \u306f\u3001Microalgae ask us Corp. (\u6c5f\u9675\u3001\u97d3\u56fd) \u304b\u3089\u5165\u624b\u3057\u307e\u3057\u305f\u3002\u30b9\u30b3\u30dd\u30e9\u30df\u30f3\u3001\u30d5\u30eb\u30aa\u30ec\u30bb\u30a4\u30f3\u3001\u30d8\u30de\u30c8\u30ad\u30b7\u30ea\u30f3\u304a\u3088\u3073\u30a8\u30aa\u30b7\u30f3 (H&E) \u306f\u3001\u52d5\u7269\u5b9f\u9a13\u7528\u306b Sigma-Aldrich (\u30bb\u30f3\u30c8\u30eb\u30a4\u30b9\u3001\u30df\u30ba\u30fc\u30ea\u5dde\u3001\u7c73\u56fd) \u304b\u3089\u8cfc\u5165\u3057\u307e\u3057\u305f\u3002\u30c0\u30eb\u30d9\u30c3\u30b3\u6539\u5909\u30a4\u30fc\u30b0\u30eb\u57f9\u5730 (DMEM):F12\u3001\u30c0\u30eb\u30d9\u30c3\u30b3\u30ea\u30f3\u9178\u7de9\u885d\u751f\u7406\u98df\u5869\u6c34 (DPBS)\u3001\u30a6\u30b7\u80ce\u5150\u8840\u6e05 (FBS)\u3001\u304a\u3088\u3073 0.25% \u30c8\u30ea\u30d7\u30b7\u30f3 - EDTA \u6eb6\u6db2\u306f\u3001Life Technologies (\u7c73\u56fd\u30ab\u30ea\u30d5\u30a9\u30eb\u30cb\u30a2\u5dde\u30ab\u30fc\u30eb\u30ba\u30d0\u30c3\u30c9) \u304b\u3089\u5165\u624b\u3057\u307e\u3057\u305f\u3002 10,000 U\/mL \u30da\u30cb\u30b7\u30ea\u30f3\u304a\u3088\u3073 10,000 \u03bcg\/mL \u30b9\u30c8\u30ec\u30d7\u30c8\u30de\u30a4\u30b7\u30f3\u3092\u542b\u3080\u6297\u751f\u7269\u8cea\u6eb6\u6db2\u306f\u3001Hyclone Laboratories Inc. (\u7c73\u56fd\u30e6\u30bf\u5dde\u30b5\u30a6\u30b9 \u30ed\u30fc\u30ac\u30f3) \u304b\u3089\u8cfc\u5165\u3057\u307e\u3057\u305f\u3002\u7d44\u63db\u3048\u30d2\u30c8 TNF-\u03b1 \u306f R&D system (\u7c73\u56fd\u30df\u30cd\u30bd\u30bf\u5dde\u30df\u30cd\u30a2\u30dd\u30ea\u30b9) \u304b\u3089\u5165\u624b\u3057\u307e\u3057\u305f\u3002\u6297\u7d30\u80de\u5916\u30b7\u30b0\u30ca\u30eb\u5236\u5fa1\u30ad\u30ca\u30fc\u30bc (ERK)\u3001\u6297 c-jun N \u672b\u7aef\u30ad\u30ca\u30fc\u30bc (JNK)\u3001\u6297 p38\u3001\u6297 \u03b1-\u30c1\u30e5\u30fc\u30d6\u30ea\u30f3\u3001\u304a\u3088\u3073\u6297 \u03b2-\u30a2\u30af\u30c1\u30f3\u3092\u542b\u3080\u4e00\u6b21\u6297\u4f53\u306f\u3001Santa Cruz Biotechnology (\u7c73\u56fd\u30ab\u30ea\u30d5\u30a9\u30eb\u30cb\u30a2\u5dde\u30b5\u30f3\u30bf\u30af\u30eb\u30fc\u30ba\uff09\u3002\u4ed6\u306e\u4e00\u6b21\u6297\u4f53\u304a\u3088\u3073\u897f\u6d0b\u308f\u3055\u3073\u30da\u30eb\u30aa\u30ad\u30b7\u30c0\u30fc\u30bc (HRP) \u7d50\u5408\u4e8c\u6b21\u6297\u4f53\u306f\u3001Cell Signaling Technology, Inc (\u7c73\u56fd\u30de\u30b5\u30c1\u30e5\u30fc\u30bb\u30c3\u30c4\u5dde\u30d3\u30d0\u30ea\u30fc) \u304b\u3089\u5165\u624b\u3057\u307e\u3057\u305f\u3002\u81ed\u5316 3-4,5-\u30b8\u30e1\u30c1\u30eb\u30c1\u30a2\u30be\u30fc\u30eb-2-\u30a4\u30eb)-2,5-\u30b8\u30d5\u30a7\u30cb\u30eb\u30c6\u30c8\u30e9\u30be\u30ea\u30a6\u30e0 (MTT) \u3092\u542b\u3080\u5316\u5b66\u7269\u8cea\u306f\u3001Sigma-Aldrich (\u30bb\u30f3\u30c8\u30eb\u30a4\u30b9\u3001\u30df\u30ba\u30fc\u30ea\u5dde\u3001\u7c73\u56fd) \u304b\u3089\u8cfc\u5165\u3057\u307e\u3057\u305f\u3002<\/p>\n

\u5316\u5b66\u5206\u6790\u6750\u6599<\/h3>\n

PUFA \u306f\u3001Sigma-Aldrich (\u7c73\u56fd\u30df\u30ba\u30fc\u30ea\u5dde\u30bb\u30f3\u30c8\u30eb\u30a4\u30b9) \u304b\u3089\u8cfc\u5165\u3057\u305f 37 \u7a2e\u985e\u306e PUFA \u6a19\u6e96\u7269\u8cea\u3092\u542b\u3080\u6eb6\u6db2\u3092\u4f7f\u7528\u3057\u3066\u540c\u5b9a\u3055\u308c\u307e\u3057\u305f\u3002\u5409\u8349\u9178\u306f\u3001\u5185\u90e8\u6a19\u6e96\u3068\u3057\u3066\u4f7f\u7528\u3059\u308b\u305f\u3081\u306b Sigma-Aldrich (\u7c73\u56fd\u30df\u30ba\u30fc\u30ea\u5dde\u30bb\u30f3\u30c8\u30eb\u30a4\u30b9) \u304b\u3089\u5165\u624b\u3057\u307e\u3057\u305f\u3002\u4e09\u30d5\u30c3\u5316\u30db\u30a6\u7d20\u30e1\u30bf\u30ce\u30fc\u30eb\u6eb6\u6db2\u3092\u542b\u3080\u6eb6\u5a92\u3001 n<\/i>-\u30d8\u30ad\u30b5\u30f3\u3001Na2<\/sub>\u305d\u308c\u30674<\/sub>\u3001CH3<\/sub>PUFA \u3092\u62bd\u51fa\u3059\u308b\u305f\u3081\u306b\u4f7f\u7528\u3055\u308c\u308b Cl\u3001MeOH\u3001\u304a\u3088\u3073 NaCl T.\u30eb\u30c6\u30a2<\/i> \u3053\u308c\u3089\u306f\u3001Sigma-Aldrich (\u7c73\u56fd\u30df\u30ba\u30fc\u30ea\u5dde\u30bb\u30f3\u30c8\u30eb\u30a4\u30b9) \u304b\u3089\u8cfc\u5165\u3057\u307e\u3057\u305f\u3002<\/p>\n

\u502b\u7406\u7684\u58f0\u660e<\/h3>\n

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\u52d5\u7269\u5b9f\u9a13\u3068DES\u30e2\u30c7\u30eb\u306e\u8a98\u5c0e<\/h3>\n

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\u7d44\u7e54\u5b66<\/h3>\n

\u89d2\u819c\u8868\u76ae\u304a\u3088\u3073\u6d99\u817a\u7d44\u7e54\u3092\u53ce\u96c6\u3057\u300110% \u30db\u30eb\u30e0\u30a2\u30eb\u30c7\u30d2\u30c9\u3067\u56fa\u5b9a\u3057\u305f\u5f8c\u3001\u30d1\u30e9\u30d5\u30a3\u30f3\u5305\u57cb\u304a\u3088\u3073\u5207\u7247\u5316\u306e\u51e6\u7406\u3092\u884c\u3044\u307e\u3057\u305f\u3002\u5207\u7247\u3092 H&E \u3067\u67d3\u8272\u3057\u3001\u9855\u5fae\u93e1 (TE-2000U\u3001\u30cb\u30b3\u30f3\u3001\u6771\u4eac\u3001\u65e5\u672c) \u3092\u4f7f\u7528\u3057\u3066 40 \u500d\u306e\u500d\u7387\u3067\u691c\u67fb\u3057\u307e\u3057\u305f\u3002\u89d2\u819c\u4e2d\u592e\u4e0a\u76ae\u306e\u539a\u3055\u306f\u3001\u5404\u89d2\u819c\u3092 5 \u3064\u306e\u30bb\u30af\u30b7\u30e7\u30f3\u306b\u5206\u5272\u3059\u308b\u3053\u3068\u306b\u3088\u3063\u3066\u6c7a\u5b9a\u3055\u308c\u307e\u3057\u305f\u3002<\/p>\n

\u514d\u75ab\u7d44\u7e54\u5316\u5b66 (IHC) \u5206\u6790\u306e\u305f\u3081\u306e DAB \u67d3\u8272<\/h3>\n

BALB\/c \u30de\u30a6\u30b9\u306e\u6d99\u817a\u304b\u3089\u7d44\u7e54\u3092\u63a1\u53d6\u3057\u300110% \u30db\u30eb\u30e0\u30a2\u30eb\u30c7\u30d2\u30c9\u3067\u56fa\u5b9a\u3057\u307e\u3057\u305f\u3002\u56fa\u5b9a\u3055\u308c\u305f\u6d99\u817a\u7d44\u7e54\u3092\u30d1\u30e9\u30d5\u30a3\u30f3\u5305\u57cb\u3057\u3001\u5207\u7247\uff085 \u03bcm\uff09\u306b\u3057\u3001\u30ad\u30b7\u30ec\u30f3\uff08Junsei\u3001\u6771\u4eac\u3001\u65e5\u672c\uff09\u3092\u4f7f\u7528\u3057\u3066\u30d1\u30e9\u30d5\u30a3\u30f3\u30923\u5206\u95933\u56de\u9664\u53bb\u3057\u307e\u3057\u305f\u3002\u5207\u7247\u3092 100\u300195\u300170\u3001\u304a\u3088\u3073 50% \u30a8\u30bf\u30ce\u30fc\u30eb\u4e2d\u3067 1 \u5206\u9593\u6c34\u548c\u3055\u305b\u307e\u3057\u305f\u3002\u6d99\u817a\u6297\u539f\u306f\u3001TRIS-EDTA \u7de9\u885d\u6db2 (pH 9.0) \u3092\u4f7f\u7528\u3057\u3066\u30de\u30a4\u30af\u30ed\u6ce2\u306b\u3088\u3063\u3066\u62bd\u51fa\u3055\u308c\u307e\u3057\u305f\u3002 3% \u904e\u9178\u5316\u6c34\u7d20\u3092\u7d44\u7e54\u30b9\u30e9\u30a4\u30c9\u306b 15 \u5206\u9593\u9069\u7528\u3057\u305f\u5f8c\u3001\u5207\u7247\u3092 2% BSA \u3067\u5ba4\u6e29\u3067 1 \u6642\u9593\u30d6\u30ed\u30c3\u30af\u3057\u307e\u3057\u305f\u3002\u5207\u7247\u3092\u3001CD45 (Bio-RAD\u3001MCA 1258GT\u30011:100 \u5e0c\u91c8) \u3092\u7528\u3044\u3066 4 \u00b0C \u3067\u4e00\u6669\u51e6\u7406\u3057\u307e\u3057\u305f\u3002 PBS \u3067\u6d17\u6d44\u3057\u305f\u5f8c\u3001\u7d30\u80de\u3092\u6297\u30a6\u30b5\u30ae IgG-HRP (Santa Cruz Biotechnology\u3001\u30c0\u30e9\u30b9\u3001\u30c6\u30ad\u30b5\u30b9\u5dde\u3001\u7c73\u56fd\u3001SC-2357\u30011:100 \u5e0c\u91c8) \u3067\u5ba4\u6e29\u3067 2 \u6642\u9593\u51e6\u7406\u3057\u30013\u3001 3'-\u30b8\u30a2\u30df\u30ce\u30d9\u30f3\u30b8\u30b8\u30f3\u56db\u5869\u9178\u5869\uff08Vector Laboratories\u3001\u30d0\u30fc\u30ea\u30f3\u30b2\u30fc\u30e0\u3001\u30ab\u30ea\u30d5\u30a9\u30eb\u30cb\u30a2\u5dde\u3001\u7c73\u56fd\uff09\u3002\u30b9\u30e9\u30a4\u30c9\u3092\u30d8\u30de\u30c8\u30ad\u30b7\u30ea\u30f3\u3067 30 \u79d2\u9593\u67d3\u8272\u3057\u3001\u6c34\u9053\u6c34\u3067\u3059\u3059\u3044\u3060\u3002\u6b21\u3044\u3067\u3001\u30b9\u30e9\u30a4\u30b9\u306b\u30ab\u30d0\u30fc\u30b9\u30ea\u30c3\u30d7\u3092\u304b\u3051\u3001Permount\u2122 mounting Medium (Thermo Fisher Scientific\u3001Waltham\u3001MA\u3001USA) \u3067\u30de\u30a6\u30f3\u30c8\u3057\u307e\u3057\u305f\u3002\u3059\u3079\u3066\u306e\u30b9\u30e9\u30a4\u30c9\u3092\u5149\u5b66\u9855\u5fae\u93e1 (Olympus\u3001CX43\u3001Olympus Optical Co.\u3001\u6771\u4eac\u3001\u65e5\u672c) \u3092\u4f7f\u7528\u3057\u3066\u691c\u67fb\u3057\u3001\u5199\u771f\u64ae\u5f71\u3057\u307e\u3057\u305f\u3002 ImageJ \u30bd\u30d5\u30c8\u30a6\u30a7\u30a2 (\u30d0\u30fc\u30b8\u30e7\u30f3 1.53\u3001\u56fd\u7acb\u885b\u751f\u7814\u7a76\u6240\u3001\u30d9\u30bb\u30b9\u30c0\u3001\u30e1\u30ea\u30fc\u30e9\u30f3\u30c9\u5dde\u3001\u7c73\u56fd)48<\/a><\/sup> \u5b9a\u91cf\u7684\u306a\u30c7\u30fc\u30bf\u5206\u6790\u3092\u5b9f\u884c\u3059\u308b\u305f\u3081\u306b\u4f7f\u7528\u3055\u308c\u307e\u3057\u305f\u3002<\/p>\n

ARPE-19 \u7d30\u80de\u57f9\u990a<\/h3>\n

\u30d2\u30c8\u7db2\u819c\u4e0a\u76ae ARPE-19 \u7d30\u80de (American Type Culture Collection\u3001ATCC\u3001\u7c73\u56fd\u30d0\u30fc\u30b8\u30cb\u30a2\u5dde\u30de\u30ca\u30b5\u30b9) \u3092\u300110% FBS (HyClone Laboratories\u3001\u30e6\u30bf\u5dde\u30ed\u30fc\u30ac\u30f3) \u3092\u6dfb\u52a0\u3057\u305f DMEM\/F-12 \u57f9\u5730 (Gibco\u3001\u7c73\u56fd\u30ab\u30ea\u30d5\u30a9\u30eb\u30cb\u30a2\u5dde\u30ab\u30fc\u30eb\u30ba\u30d0\u30c3\u30c9) \u3067\u57f9\u990a\u3057\u307e\u3057\u305f\u3002 \u3001\u7c73\u56fd\uff09\u304a\u3088\u3073 1% \u30da\u30cb\u30b7\u30ea\u30f3\/\u30b9\u30c8\u30ec\u30d7\u30c8\u30de\u30a4\u30b7\u30f3\uff08HyClone Laboratories\uff09\u3002\u7d30\u80de\u306f 2 \u00d7 10 \u500b\u306e\u5bc6\u5ea6\u3067\u64ad\u7a2e\u3055\u308c\u307e\u3057\u305f\u30025<\/sup> 6\u30a6\u30a7\u30eb\u30d7\u30ec\u30fc\u30c8\u306e\u30a6\u30a7\u30eb\u3054\u3068\u306b\u57f9\u990a\u3057\u3001\u708e\u75c7\u53cd\u5fdc\u304a\u3088\u3073\u5c0f\u80de\u4f53\u53cd\u5fdc\u3092\u8a98\u5c0e\u3057\u307e\u3059\u3002<\/p>\n

\u7d30\u80de\u751f\u5b58\u7387<\/h3>\n

\u306e\u52b9\u679c T.\u30eb\u30c6\u30a2<\/i> \u7d30\u80de\u751f\u5b58\u7387\u306f\u3001\u4ee5\u524d\u306e\u7814\u7a76\u306b\u8a18\u8f09\u3055\u308c\u3066\u3044\u308b\u3088\u3046\u306b\u3001MTT \u30a2\u30c3\u30bb\u30a4\u306b\u3088\u3063\u3066\u8a55\u4fa1\u3055\u308c\u307e\u3057\u305f\u300249<\/a><\/sup> \u305f\u3060\u3057\u3001\u308f\u305a\u304b\u306a\u5909\u66f4\u304c\u52a0\u3048\u3089\u308c\u3066\u3044\u307e\u3059\u3002\u7d30\u80de\u3092 1.5 \u00d7 10 \u306e\u5bc6\u5ea6\u3067 96 \u30a6\u30a7\u30eb\u57f9\u990a\u30d7\u30ec\u30fc\u30c8\u306b\u64ad\u7a2e\u3057\u307e\u3057\u305f\u30024<\/sup> \u7d30\u80de\/\u30a6\u30a7\u30eb\u306724\u6642\u9593\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30c8\u3057\u307e\u3057\u305f\u3002\u6b21\u306b\u3001\u7d30\u80de\u3092\u3055\u307e\u3056\u307e\u306a\u6fc3\u5ea6\u306e T.\u30eb\u30c6\u30a2<\/i> \u3055\u3089\u306b24\u6642\u9593\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30c8\u3057\u307e\u3057\u305f\u3002\u5404\u30a6\u30a7\u30eb\u306e\u4e0a\u6e05\u3092\u30010.5 mg\/mL \u306e MTT \u3092\u542b\u3080\u65b0\u9bae\u306a\u5897\u6b96\u57f9\u5730\u3068\u4ea4\u63db\u3057\u30011 \u6642\u9593\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30c8\u3057\u307e\u3057\u305f\u3002\u4e0a\u6e05\u3092\u9664\u53bb\u3057\u3001\u751f\u6210\u3057\u305f\u30db\u30eb\u30de\u30b6\u30f3\u6c88\u6bbf\u7269\u3092\uff12\uff10\uff10\u03bc\uff2c\u306e\u30b8\u30e1\u30c1\u30eb\u30b9\u30eb\u30db\u30ad\u30b7\u30c9\u306b\u6eb6\u89e3\u3057\u305f\u3002\u5438\u5149\u5ea6\u306f570nm\u3067\u6e2c\u5b9a\u3057\u305f\u3002\u7d30\u80de\u751f\u5b58\u7387\u306f\u3001\u9670\u6027\u5bfe\u7167\u7fa4\u3068\u6bd4\u8f03\u3057\u305f\u30d1\u30fc\u30bb\u30f3\u30c6\u30fc\u30b8\u3068\u3057\u3066\u76f8\u5bfe\u7684\u306b\u8a08\u7b97\u3055\u308c\u307e\u3057\u305f\u3002<\/p>\n

\u30a6\u30a7\u30b9\u30bf\u30f3\u30d6\u30ed\u30c3\u30c8\u5206\u6790<\/h3>\n

\u30a6\u30a7\u30b9\u30bf\u30f3\u30d6\u30ed\u30c3\u30c6\u30a3\u30f3\u30b0\u306f\u3001\u6297\u708e\u75c7\u4f5c\u7528\u306e\u6839\u5e95\u306b\u3042\u308b\u7d30\u80de\u30b7\u30b0\u30ca\u30eb\u4f1d\u9054\u7d4c\u8def\u3092\u6c7a\u5b9a\u3059\u308b\u305f\u3081\u306b\u5b9f\u65bd\u3055\u308c\u307e\u3057\u305f\u3002 T.\u30eb\u30c6\u30a2<\/i> \u524d\u8ff0\u3057\u305f\u3088\u3046\u306b\u7814\u7a7649<\/a><\/sup>\u3001\u305f\u3060\u3057\u5c0f\u3055\u306a\u5909\u66f4\u304c\u52a0\u3048\u3089\u308c\u3066\u3044\u307e\u3059\u3002\u30b3\u30f3\u30d5\u30eb\u30a8\u30f3\u30c8\u306a ARPE-19 \u7d30\u80de\u3092\u30011% FBS \u304a\u3088\u3073 1% \u6297\u751f\u7269\u8cea\u3092\u542b\u3080 DMEM:F12 \u4e2d\u3067 24 \u6642\u9593\u98e2\u9913\u72b6\u614b\u306b\u3057\u307e\u3057\u305f\u3002\u6b21\u3044\u3067\u3001\u7d30\u80de\u3092\u6b21\u306e\u3082\u306e\u3067\u51e6\u7406\u3057\u305f\u3002 T.\u30eb\u30c6\u30a2<\/i> (50 \u304a\u3088\u3073 100 \u03bcg\/mL) \u3067 2 \u6642\u9593\u523a\u6fc0\u3057\u3001\u305d\u306e\u5f8c 20 ng\/mL \u306e TNF-\u03b1 \u3067 15 \u5206\u9593\u523a\u6fc0\u3057\u307e\u3057\u305f\u3002\u7d30\u80de\u3092\u6c37\u51b7\u3057\u305fDPBS\u30673\u56de\u6d17\u6d44\u3057\u3001\u30d7\u30ed\u30c6\u30a2\u30fc\u30bc\u304a\u3088\u3073\u30db\u30b9\u30d5\u30a1\u30bf\u30fc\u30bc\u963b\u5bb3\u5264\u306e\u30ab\u30af\u30c6\u30eb\u3092\u6dfb\u52a0\u3057\u305f\u30bf\u30f3\u30d1\u30af\u8cea\u62bd\u51fa\u7de9\u885d\u6db2(iNtRON Biotechnology\u3001\u4eac\u757f\u9053\u3001\u97d3\u56fd)\u4e2d\u3067\u6eb6\u89e3\u3057\u305f\u3002\u30e9\u30a4\u30bb\u30fc\u30c8\u306e\u4e0a\u6e05\u3092\u9060\u5fc3\u5206\u96e2 (14,000\u00d7) \u306b\u3088\u3063\u3066\u53d6\u5f97\u3057\u307e\u3057\u305f\u3002g<\/i>\u300130\u5206\u30014\u2103\uff09\u3001\u305d\u308c\u3089\u306e\u30bf\u30f3\u30d1\u30af\u8cea\u6fc3\u5ea6\u306fDCTM\u30bf\u30f3\u30d1\u30af\u8cea\u30a2\u30c3\u30bb\u30a4\u30ad\u30c3\u30c8\uff08Bio-Rad\u3001Hercules\u3001CA\u3001USA\uff09\u3092\u4f7f\u7528\u3057\u3066\u63a8\u5b9a\u3055\u308c\u307e\u3057\u305f\u3002\u7b49\u91cf (15 \uff5e 25 \u03bcg) \u306e\u7d30\u80de\u30bf\u30f3\u30d1\u30af\u8cea\u3092 SDS-PAGE \u3067\u5206\u96e2\u3057\u3001PVDF \u819c\u306b\u8ee2\u5199\u3057\u307e\u3057\u305f\u3002\u819c\u30925%\u30a6\u30b7\u8840\u6e05\u30a2\u30eb\u30d6\u30df\u30f3\u307e\u305f\u306f\u30b9\u30ad\u30e0\u30df\u30eb\u30af\u3068\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30c8\u3057\u3001\u6b21\u306b1:2000\u5e0c\u91c8\u306e\u4e00\u6b21\u6297\u4f53\uff08\u6297\u30db\u30b9\u30dbERK\u3001\u6297ERK\u3001\u6297\u30db\u30b9\u30dbJNK\u3001\u6297JNK\u3001\u6297\u30db\u30b9\u30dbp38\uff09\u3068\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30c8\u3057\u307e\u3057\u305f\u3002 \u3001\u6297 p38\u3001\u6297\u30db\u30b9\u30db AKT\u3001\u6297 AKT\u3001\u6297\u30db\u30b9\u30db I\u03baB-\u03b1\u3001\u6297 I\u03baB-\u03b1\u3001\u6297\u30db\u30b9\u30db p65\u3001\u6297 p65\u3001\u6297 \u03b1-\u30c1\u30e5\u30fc\u30d6\u30ea\u30f3\u304a\u3088\u3073\u6297 \u03b2-\u30a2\u30af\u30b7\u30e7\u30f3) 4 \u00b0C \u3067 12 \u6642\u9593\u3002\u6b21\u3044\u3067\u3001\u819c\u3092TBS-T\u3067\u3059\u3059\u304e\u30011:10,000\u306e\u5e0c\u91c8\u306e\u6297\u30a6\u30b5\u30aeHRP\u7d50\u5408\u4e8c\u6b21\u6297\u4f53\u3067\u5ba4\u6e29\u30671\u6642\u9593\u51e6\u7406\u3057\u305f\u3002 SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Fisher Scientific) \u3092\u4f7f\u7528\u3057\u3066\u30bf\u30f3\u30d1\u30af\u8cea\u30d0\u30f3\u30c9\u3092\u691c\u51fa\u3057\u3001iBright CL1000 \u30b2\u30eb \u30c9\u30ad\u30e5\u30e1\u30f3\u30c6\u30fc\u30b7\u30e7\u30f3 \u30b7\u30b9\u30c6\u30e0 (Thermo Fisher Scientific) \u3067\u8996\u899a\u5316\u3057\u307e\u3057\u305f\u3002\u5404\u30bf\u30f3\u30d1\u30af\u8cea\u30d6\u30ed\u30c3\u30c8\u306e\u5bc6\u5ea6\u306f\u3001Image J \u30bd\u30d5\u30c8\u30a6\u30a7\u30a2 (\u30d0\u30fc\u30b8\u30e7\u30f3 1.53\u3001NIH) \u3092\u4f7f\u7528\u3057\u3066\u6e2c\u5b9a\u3057\u307e\u3057\u305f\u300248<\/a><\/sup>.<\/p>\n

\u7dcfPUFA\u62bd\u51fa\u91cf T.\u30eb\u30c6\u30a2<\/i><\/h3>\n

\u7dcfPUFA\u306f\u4ee5\u4e0b\u304b\u3089\u62bd\u51fa\u3055\u308c\u307e\u3057\u305f T.\u30eb\u30c6\u30a2<\/i> \u30d5\u30a9\u30eb\u30c1\u30c6\u30af\u30cb\u30c3\u30af\u3092\u4f7f\u3063\u306650<\/a><\/sup>\u3002\u7c21\u5358\u306b\u8aac\u660e\u3059\u308b\u3068\u3001100 mg \u306e\u30b5\u30f3\u30d7\u30eb\u3092 5 mL \u306e CH \u304c\u5165\u3063\u305f\u30d0\u30a4\u30a2\u30eb\u306b\u5165\u308c\u307e\u3057\u305f\u30023<\/sub>Cl:MeOH (2:1\u3001v\/v) \u6eb6\u6db2\u3092\u52a0\u3048\u300115 \u5206\u9593\u8d85\u97f3\u6ce2\u51e6\u7406\u3057\u307e\u3057\u305f\u3002\u61f8\u6fc1\u6db2\u309210,000 rpm\u30675\u5206\u9593\u9060\u5fc3\u5206\u96e2\u3057\u3066\u6e05\u6f84\u5316\u3057\u305f\u3002\u30ca2<\/sub>\u305d\u308c\u30674<\/sub> \u3092\u4e0a\u6e05\u306b\u52a0\u3048\u3001\u6b21\u3044\u3067\u6ffe\u904e\u3057\u3066\u6b8b\u7559\u6c34\u3092\u9664\u53bb\u3057\u305f\u3002\u3053\u306e\u30b9\u30c6\u30c3\u30d7\u3092\u3055\u3089\u306b 2 \u56de\u7e70\u308a\u8fd4\u3057\u307e\u3057\u305f\u3002\u6b21\u3044\u3067\u3001\u6eb6\u6db2\u3092\u7a92\u7d20\u30ac\u30b9\u4e0b\u3067\u6fc3\u7e2e\u3057\u3066\u3001\u7d14\u7c8b\u306a\u85fb\u985e\u8102\u8cea\u3092\u5f97\u305f\u3002<\/p>\n

\u8102\u80aa\u9178\u30e1\u30c1\u30eb\u5316<\/h3>\n

\u8102\u80aa\u9178\u30e1\u30c1\u30eb\u5316\u3092\u5b9f\u884c\u3057\u3066\u3001\u30ac\u30b9\u30af\u30ed\u30de\u30c8\u30b0\u30e9\u30d5\u30a3\u30fc (GC) \u5206\u6790\u7528\u306e\u8102\u8cea\u30b5\u30f3\u30d7\u30eb\u3092\u8abf\u88fd\u3057\u307e\u3057\u305f\u3002\u62bd\u51fa\u3057\u305f\u7269\u8cea\u309210 mL\u30d0\u30a4\u30a2\u30eb\u306b\u52a0\u3048\u3001\u7d9a\u3044\u30661 mL\u306e0.5 N NaOH:MeOH\u6df7\u5408\u7269\u3092\u52a0\u3048\u305f\u3002\u3053\u306e\u6eb6\u6db2\u306b\u306f 400 \u03bcg\/mL \u306e\u5185\u90e8\u30b9\u30c8\u30e9\u30f3\u30c9 (\u5409\u8349\u9178) \u304c\u542b\u307e\u308c\u3066\u3044\u307e\u3059\u3002\u6df7\u5408\u7269\u3092 30 \u79d2\u9593\u632f\u76ea\u3057\u3001\u305d\u306e\u5f8c 80 \u00b0C \u3067 20 \u5206\u9593\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30c8\u3057\u307e\u3057\u305f\u3002\u6b21\u3044\u3067\u3001\u6df7\u5408\u7269\u3092\u5ba4\u6e29\u3067\uff15\u5206\u9593\u51b7\u5374\u3057\u3001\u6b21\u3044\u3067\uff12\uff4d\uff2c\u306e\u4e09\u30d5\u30c3\u5316\u30db\u30a6\u7d20\uff1a\uff2d\uff45\uff2f\uff28\u3092\u6dfb\u52a0\u3057\u3001\u6b21\u3044\u3067\uff13\uff10\u79d2\u9593\u64b9\u62cc\u3057\u305f\u3002\u6df7\u5408\u7269\u309280\u2103\u306720\u5206\u9593\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30c8\u3057\u3001\u305d\u306e\u5f8c10\u5206\u9593\u51b7\u5374\u3057\u305f\u3002 2 mL \u306e\u904e\u98fd\u548c NaCl \u3092\u6dfb\u52a0\u3057\u3066\u3001\u76f8\u5206\u96e2\u3068\u8102\u80aa\u9178\u306e\u53ce\u96c6\u3092\u8a98\u5c0e\u3057\u307e\u3057\u305f\u3002\u6b21\u306b\u30011.5mL n<\/i>\u2212\u30d8\u30ad\u30b5\u30f3\u3092\u52a0\u3048\u3001\u6df7\u5408\u7269\u3092\uff13\uff10\u79d2\u9593\u30dc\u30eb\u30c6\u30c3\u30af\u30b9\u3057\u305f\u5f8c\u3001\u5ba4\u6e29\u3067\uff12\uff10\u5206\u9593\u51b7\u5374\u3057\u305f\u3002\u5c64\u3092\u5206\u96e2\u3057\u3001\u6db2\u4f53\u4e0a\u6f84\u307f\u3092\u6df7\u5408\u7269\u304b\u3089\u9664\u53bb\u3057\u3001\u98fd\u548cNaHCO \u3092\u4f7f\u7528\u3057\u3066\u6c34\u3092\u5b8c\u5168\u306b\u9664\u53bb\u3057\u305f\u30022<\/sub>\u305d\u308c\u30674<\/sub> \u30d5\u30a3\u30eb\u30bf\u30fc\u3002<\/p>\n

\u8102\u80aa\u9178\u306e\u540c\u5b9a<\/h3>\n

\u8102\u80aa\u9178\u30e1\u30c1\u30eb\u30a8\u30b9\u30c6\u30eb (FAME) \u306f\u3001\u708e\u30a4\u30aa\u30f3\u5316\u691c\u51fa\u5668 (FID; Agilent 7890A\u3001Agilent Technologies\u3001\u7c73\u56fd\u30c7\u30e9\u30a6\u30a7\u30a2\u5dde\u30a6\u30a3\u30eb\u30df\u30f3\u30c8\u30f3) \u3092\u5099\u3048\u305f GC \u3092\u4f7f\u7528\u3057\u3066\u691c\u67fb\u3057\u307e\u3057\u305f\u3002\u30b5\u30f3\u30d7\u30eb (1 \u03bcL) \u3092 GC \u306e HP-88 \u30ab\u30e9\u30e0 (J&W 112-88A7\u3001Agilent Technologies\u3001\u30a6\u30a3\u30eb\u30df\u30f3\u30c8\u30f3\u3001\u30c7\u30e9\u30a6\u30a7\u30a2\u5dde\u3001\u7c73\u56fd) (100 m \u00d7 250 \u03bcm ID\u30010.2 \u03bcm \u30d5\u30a3\u30eb\u30e0) \u306b\u6ce8\u5165\u3057\u307e\u3057\u305f\u3002 GC \u5206\u6790\u306f\u6b21\u306e\u3088\u3046\u306b\u5b9f\u884c\u3055\u308c\u307e\u3057\u305f\u3002\u30b5\u30f3\u30d7\u30eb\u306f\u6700\u521d\u306b 140 \u00b0C \u3067 5 \u5206\u9593\u4fdd\u6301\u3055\u308c\u3001\u6b21\u306b\u6e29\u5ea6\u3092 4 \u00b0C\/min \u3067 15 \u5206\u9593\u304b\u3051\u3066 240 \u00b0C \u307e\u3067\u4e0a\u6607\u3055\u305b\u307e\u3057\u305f\u3002\u30ab\u30e9\u30e0\u306e\u6d41\u901f\u306f 1 mL\/min \u306b\u56fa\u5b9a\u3057\u307e\u3057\u305f\u3002\u5206\u6790\u7528\u306e\u30ad\u30e3\u30ea\u30a2\u30ac\u30b9\u3068\u3057\u3066\u7a92\u7d20\u30ac\u30b9\u3092\u4f7f\u7528\u3057\u3001FID \u3092 280 \u00b0C \u306b\u8a2d\u5b9a\u3057\u3001\u30b9\u30d7\u30ea\u30c3\u30c8\u6bd4\u3092 30:1 \u306b\u8a2d\u5b9a\u3057\u307e\u3057\u305f\u3002 FAME \u306f\u3001\u65e2\u77e5\u306e\u6a19\u6e96\u3068\u6bd4\u8f03\u3059\u308b\u3053\u3068\u306b\u3088\u3063\u3066\u7279\u5b9a\u3055\u308c\u307e\u3057\u305f\u3002\u5404\u30b5\u30f3\u30d7\u30eb\u3092 GC \u5206\u6790\u306b\u3088\u308a 3 \u56de\u6e2c\u5b9a\u3057\u307e\u3057\u305f\u3002<\/p>\n

\u7d71\u8a08\u5206\u6790<\/h3>\n

\u3059\u3079\u3066\u306e\u30c7\u30fc\u30bf\u306f\u3001SPSS \u30d0\u30fc\u30b8\u30e7\u30f3 18.0 (IBM Co.\u3001\u7c73\u56fd\u30cb\u30e5\u30fc\u30e8\u30fc\u30af\u5dde\u30a2\u30fc\u30e2\u30f3\u30af) \u306b\u3088\u308b Tukey \u306e\u4e8b\u5f8c\u5206\u6790\u3092\u4f34\u3046\u4e00\u5143\u914d\u7f6e\u5206\u6563\u5206\u6790\u3092\u4f7f\u7528\u3057\u3066\u7d71\u8a08\u7684\u306b\u5206\u6790\u3055\u308c\u307e\u3057\u305f\u3002\u5404\u5b9f\u9a13\u306b\u3064\u3044\u3066\u3001\u5c11\u306a\u304f\u3068\u3082 3 \u3064\u306e\u72ec\u7acb\u3057\u305f\u53cd\u5fa9\u304c\u5b9f\u884c\u3055\u308c\u307e\u3057\u305f\u3002<\/p>\n<\/div>\n

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\u30bd\u30fc\u30b9\u30ea\u30f3\u30af <\/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\/ja\/wp-json\/wp\/v2\/posts\/4903","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/directcm.com\/ja\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/directcm.com\/ja\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/directcm.com\/ja\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/directcm.com\/ja\/wp-json\/wp\/v2\/comments?post=4903"}],"version-history":[{"count":1,"href":"https:\/\/directcm.com\/ja\/wp-json\/wp\/v2\/posts\/4903\/revisions"}],"predecessor-version":[{"id":5393,"href":"https:\/\/directcm.com\/ja\/wp-json\/wp\/v2\/posts\/4903\/revisions\/5393"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/directcm.com\/ja\/wp-json\/wp\/v2\/media\/5363"}],"wp:attachment":[{"href":"https:\/\/directcm.com\/ja\/wp-json\/wp\/v2\/media?parent=4903"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/directcm.com\/ja\/wp-json\/wp\/v2\/categories?post=4903"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/directcm.com\/ja\/wp-json\/wp\/v2\/tags?post=4903"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}