{"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\/zh\/protective-effect-of-tisochrysis-lutea-on-dry-eye-syndrome-via-nf-%ce%bab-inhibition\/","title":{"rendered":"\u9ec4\u91d1\u85fb\u901a\u8fc7\u6291\u5236 NF-\u03baB \u5bf9\u5e72\u773c\u7efc\u5408\u5f81\u7684\u4fdd\u62a4\u4f5c\u7528"},"content":{"rendered":"
\n

\u6750\u6599<\/h3>\n

\u4f53\u5185\/\u4f53\u5916\u6750\u6599<\/h4>\n

\u9ec4\u6bdb\u866b<\/i> \u83b7\u81eaMicroalgae Ask us Corp.\uff08\u97e9\u56fd\u6c5f\u9675\uff09\u3002\u4e1c\u83a8\u83ea\u78b1\u3001\u8367\u5149\u7d20\u3001\u82cf\u6728\u7cbe\u548c\u4f0a\u7ea2 (H&E) \u8d2d\u81ea Sigma-Aldrich\uff08\u7f8e\u56fd\u5bc6\u82cf\u91cc\u5dde\u5723\u8def\u6613\u65af\uff09\uff0c\u7528\u4e8e\u52a8\u7269\u5b9e\u9a8c\u3002\u675c\u5c14\u8d1d\u79d1\u6539\u826f\u4f0a\u683c\u5c14\u57f9\u517b\u57fa (DMEM)\uff1aF12\u3001\u675c\u5c14\u8d1d\u79d1\u78f7\u9178\u76d0\u7f13\u51b2\u76d0\u6c34 (DPBS)\u3001\u80ce\u725b\u8840\u6e05 (FBS) \u548c 0.25% \u80f0\u86cb\u767d\u9176-EDTA \u6eb6\u6db2\u83b7\u81ea Life Technologies\uff08\u5361\u5c14\u65af\u5df4\u5fb7\uff0c\u52a0\u5229\u798f\u5c3c\u4e9a\u5dde\uff0c\u7f8e\u56fd\uff09\u3002\u6297\u751f\u7d20\u6eb6\u6db2\u5305\u542b 10,000 U\/mL \u9752\u9709\u7d20\u548c 10,000 \u03bcg\/mL \u94fe\u9709\u7d20\uff0c\u8d2d\u81ea Hyclone Laboratories Inc.\uff08\u7f8e\u56fd\u72b9\u4ed6\u5dde\u5357\u6d1b\u6839\uff09\u3002\u91cd\u7ec4\u4eba TNF-\u03b1 \u83b7\u81ea\u7814\u53d1\u7cfb\u7edf\uff08\u660e\u5c3c\u82cf\u8fbe\u5dde\u660e\u5c3c\u963f\u6ce2\u5229\u65af\u5e02\uff0c\u7f8e\u56fd\uff09\u3002\u4e00\u6297\u5305\u62ec\u6297\u7ec6\u80de\u5916\u4fe1\u53f7\u8c03\u8282\u6fc0\u9176\uff08ERK\uff09\u3001\u6297c-jun N\u672b\u7aef\u6fc0\u9176\uff08JNK\uff09\u3001\u6297p38\u3001\u6297\u03b1-\u5fae\u7ba1\u86cb\u767d\u548c\u6297\u03b2-\u808c\u52a8\u86cb\u767d\uff0c\u8d2d\u81eaSanta Cruz Biotechnology\uff08\u7f8e\u56fd\u52a0\u5229\u798f\u5c3c\u4e9a\u5dde\u5723\u514b\u9c81\u65af\uff09\u3002\u5176\u4ed6\u4e00\u6297\u548c\u8fa3\u6839\u8fc7\u6c27\u5316\u7269\u9176 (HRP) \u7f00\u5408\u7684\u4e8c\u6297\u8d2d\u81ea Cell Signaling Technology, Inc\uff08\u7f8e\u56fd\u9a6c\u8428\u8bf8\u585e\u5dde\u8d1d\u5f17\u5229\uff09\u3002\u5305\u62ec 3\u20134,5-\u4e8c\u7532\u57fa\u567b\u5511-2-\u57fa)-2,5-\u4e8c\u82ef\u57fa\u6eb4\u5316\u56db\u5511 (MTT) \u5728\u5185\u7684\u5316\u5b66\u54c1\u8d2d\u81ea Sigma-Aldrich\uff08\u7f8e\u56fd\u5bc6\u82cf\u91cc\u5dde\u5723\u8def\u6613\u65af\uff09\u3002<\/p>\n

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

PUFA \u4f7f\u7528\u542b\u6709 37 \u79cd PUFA \u6807\u51c6\u7269\u8d28\u7684\u6eb6\u6db2\u8fdb\u884c\u9274\u5b9a\uff0c\u8be5\u6eb6\u6db2\u8d2d\u81ea Sigma-Aldrich\uff08\u7f8e\u56fd\u5bc6\u82cf\u91cc\u5dde\u5723\u8def\u6613\u65af\uff09\u3002\u620a\u9178\u8d2d\u81ea Sigma-Aldrich\uff08\u7f8e\u56fd\u5bc6\u82cf\u91cc\u5dde\u5723\u8def\u6613\u65af\uff09\uff0c\u7528\u4f5c\u5185\u6807\u3002\u6eb6\u5242\uff0c\u5305\u62ec\u4e09\u6c1f\u5316\u787c\u7532\u9187\u6eb6\u6db2\uff0c n<\/i>-\u5df1\u70f7\u3001Na2<\/sub>\u6240\u4ee54<\/sub>,CH3<\/sub>Cl\u3001MeOH \u548c NaCl\uff0c\u7528\u4e8e\u4ece\u5176\u4e2d\u63d0\u53d6 PUFA \u9ec4\u6bdb\u866b<\/i> \u8d2d\u81ea Sigma-Aldrich\uff08\u7f8e\u56fd\u5bc6\u82cf\u91cc\u5dde\u5723\u8def\u6613\u65af\uff09\u3002<\/p>\n

\u9053\u5fb7\u58f0\u660e<\/h3>\n

\u8fd9\u9879\u7814\u7a76\u5f97\u5230\u4e86\u97e9\u56fd\u79d1\u5b66\u6280\u672f\u7814\u7a76\u9662 (KIST) \u673a\u6784\u52a8\u7269\u62a4\u7406\u548c\u4f7f\u7528\u59d4\u5458\u4f1a (IACUC) \u7684\u6279\u51c6\uff1aKIST No. 2020-002\u3001\u6c5f\u9675\u7814\u7a76\u6240\uff0c\u5e76\u9075\u5faa\u89c6\u89c9\u548c\u773c\u79d1\u7814\u7a76\u534f\u4f1a\u5173\u4e8e\u52a8\u7269\u5728\u773c\u79d1\u548c\u89c6\u529b\u7814\u7a76\u4e2d\u7684\u4f7f\u7528\u3002\u6b64\u5916\uff0c\u6211\u4eec\u9075\u5faa\u52a8\u7269\u7814\u7a76\uff1a\u4f53\u5185\u5b9e\u9a8c\u62a5\u544a (ARRIVE) \u5efa\u8bae\u3002\u6240\u6709\u7a0b\u5e8f\u5747\u6309\u7167\u9002\u7528\u7684\u6cd5\u89c4\u548c\u51c6\u5219\u8fdb\u884c\u3002<\/p>\n

\u52a8\u7269\u5b9e\u9a8c\u53caDES\u6a21\u578b\u8bf1\u5bfc<\/h3>\n

\u5c06\u5c0f\u9f20\u9972\u517b\u5728\u88c5\u6709\u7a7a\u8c03\u7684\u52a8\u7269\u623f\u4e2d\uff0c\u7ef4\u6301\u572822\u2103\u00b12\u2103\u300150%\u00b110%\u6e7f\u5ea6\u548c12\u5c0f\u65f6\u5149\u7167\/12\u5c0f\u65f6\u9ed1\u6697\u663c\u591c\u8282\u5f8b\u5468\u671f\u3002\u968f\u610f\u63d0\u4f9b\u98df\u7269\u548c\u6c34\u3002\u5c0f\u9f20\u9002\u5e94\u4e00\u5468\u540e\u88ab\u968f\u673a\u5206\u4e3a\u4e94\u7ec4\uff0c\u6bcf\u7ec4\u4e03\u53ea 6 \u5468\u9f84\u96c4\u6027 BALB\/c \u5c0f\u9f20\uff08Orientbio\uff0cGyunggi-do\uff0c\u97e9\u56fd\uff09\u3002\u901a\u8fc7\u8179\u8154\u6ce8\u5c04 200 \u03bcL (2.5 mg\/mL) \u7a00\u91ca\u4e8e\u78f7\u9178\u76d0\u7f13\u51b2\u76d0\u6c34\uff08PBS\uff1bSigma-Aldrich, St. Louis, MO, USA\uff09\u7684\u4e1c\u83a8\u83ea\u78b1\uff0c\u5728\u5c0f\u9f20\u4e2d\u5b9e\u9a8c\u8bf1\u5bfc DES\uff0c\u6bcf\u5929\u4e24\u6b21\u3002 \u9ec4\u6bdb\u866b<\/i> \u6bcf\u5929\u4ee5 0\uff08\u8f7d\u4f53\u5bf9\u7167\uff09\u3001100\u3001150 \u6216 300 mg\/kg \u7684\u6d53\u5ea6\uff08\u6eb6\u4e8e 200 \u03bcL \u84b8\u998f\u6c34\uff09\u53e3\u670d\u7ed9\u4e88\u5c0f\u9f20\u7ec4\u3002\u5bf9\u7167\u7ec4\u63a5\u53d7\u4e0d\u542b\u4e1c\u83a8\u83ea\u78b1\u7684PBS\u3002\u4ec0\u4e48\u65f6\u5019 \u9ec4\u6bdb\u866b<\/i> \u53e3\u670d\u7ed9\u836f\uff0c\u5bf9\u7167\u7ec4\u548cDES\u7ec4\u4f7f\u7528200\u03bcL\u84b8\u998f\u6c34\u3002\u4e24\u5468\u540e\uff0c\u5728\u95ed\u773c 1 \u5206\u949f\u4e4b\u524d\uff0c\u4f7f\u7528\u653e\u7f6e\u5728\u989e\u773c\u7751\u4e0b\u4e09\u5206\u4e4b\u4e00\u5904\u7684\u6807\u51c6 Schirmer \u6d4b\u8bd5\u6761\u5bf9\u6cea\u6db2\u4ea7\u751f\u8fdb\u884c\u91cf\u5316\u3002\u7136\u540e\u53d6\u51fa\u6761\u5e26\u5e76\u6d4b\u91cf\u6e7f\u70b9\u7684\u957f\u5ea6\uff08\u4ee5\u6beb\u7c73\u4e3a\u5355\u4f4d\uff09\u4ee5\u786e\u5b9a\u5e2d\u5c14\u9ed8\u7684\u6d4b\u8bd5\u503c\u3002\u89d2\u819c\u8868\u9762\u7684\u67d3\u8272\u7528\u4e8e\u786e\u5b9a\u89d2\u819c\u8868\u9762\u53d8\u5316\u7684\u7a0b\u5ea6\u3002\u4e0b\u5916\u4fa7\u7ed3\u819c\u6ce8\u5c041\u6ef43%\u8367\u5149\u7d20\u540e\u89c2\u5bdf\u89d2\u819c\u8868\u9762\u5e76\u8bc4\u5206\u3002\u76f2\u76ee\u8bc4\u4f30\u89d2\u819c\u67d3\u8272\u3002\u901a\u8fc7\u9888\u690e\u8131\u4f4d\u5904\u6b7b\u5c0f\u9f20\u3002\u901a\u8fc7\u8f7b\u5f39\u5c0f\u9f20\u773c\u7751\u5e76\u4f7f\u7528\u954a\u5b50\u4ece\u773c\u7403\u5207\u51fa\u89c6\u795e\u7ecf\u5e76\u53bb\u9664\u89d2\u819c\u6765\u53bb\u9664\u89d2\u819c\u3002\u5c06\u6676\u72b6\u4f53\u4ece\u89d2\u819c\u5185\u53d6\u51fa\u5e76\u4fdd\u5b58\u5728\u221280\u00b0C \u4e0b\u3002\u4f7f\u7528\u954a\u5b50\u4ece\u5c0f\u9f20\u7684\u4e0b\u988c\u53d6\u51fa\u6cea\u817a\uff0c\u5e76\u5207\u6389\u62c9\u52a8\u533a\u57df\u7684\u8868\u76ae\u4ee5\u9732\u51fa\u4e0b\u534a\u5f20\u8138\u3002\u56fa\u5b9a\u5e76\u53d6\u51fa\u4e0b\u988c\u9644\u8fd1\u7684\u6cea\u817a\uff0c\u7136\u540e\u907f\u5149\u4fdd\u5b58\u5728\u2212 80\u00b0C \u4e0b\u3002<\/p>\n

\u7ec4\u7ec7\u5b66<\/h3>\n

\u6536\u96c6\u89d2\u819c\u8868\u76ae\u548c\u6cea\u817a\u7ec4\u7ec7\u5e76\u56fa\u5b9a\u572810%\u7532\u919b\u4e2d\uff0c\u7136\u540e\u8fdb\u884c\u77f3\u8721\u5305\u57cb\u548c\u5207\u7247\u5904\u7406\u3002\u5207\u7247\u7528 H&E \u67d3\u8272\uff0c\u5e76\u4f7f\u7528\u663e\u5fae\u955c\uff08TE-2000U\uff0c\u5c3c\u5eb7\uff0c\u4e1c\u4eac\uff0c\u65e5\u672c\uff09\u5728 40 \u500d\u653e\u5927\u500d\u6570\u4e0b\u8fdb\u884c\u68c0\u67e5\u3002\u901a\u8fc7\u5c06\u6bcf\u4e2a\u89d2\u819c\u5206\u4e3a\u4e94\u4e2a\u90e8\u5206\u6765\u786e\u5b9a\u4e2d\u592e\u89d2\u819c\u4e0a\u76ae\u539a\u5ea6\u3002<\/p>\n

\u7528\u4e8e\u514d\u75ab\u7ec4\u7ec7\u5316\u5b66 (IHC) \u5206\u6790\u7684 DAB \u67d3\u8272<\/h3>\n

\u53d6BALB\/c\u5c0f\u9f20\u6cea\u817a\u7ec4\u7ec7\u5e76\u752810%\u7532\u919b\u56fa\u5b9a\u3002\u5c06\u56fa\u5b9a\u7684\u6cea\u817a\u7ec4\u7ec7\u77f3\u8721\u5305\u57cb\uff0c\u5207\u7247\uff085\u03bcm\uff09\uff0c\u5e76\u4f7f\u7528\u4e8c\u7532\u82ef\u53bb\u9664\u77f3\u87213\u6b21\uff0c\u6bcf\u6b213\u5206\u949f\uff08Junsei\uff0cTokyo\uff0cJapan\uff09\u3002\u5c06\u5207\u7247\u5728 100\u300195\u300170 \u548c 50% \u4e59\u9187\u4e2d\u6c34\u5408 1 \u5206\u949f\u3002\u4f7f\u7528 TRIS-EDTA \u7f13\u51b2\u6db2\uff08pH 9.0\uff09\u5fae\u6ce2\u63d0\u53d6\u6cea\u817a\u6297\u539f\u3002\u5c06 3% \u8fc7\u6c27\u5316\u6c22\u5e94\u7528\u4e8e\u7ec4\u7ec7\u73bb\u7247 15 \u5206\u949f\u540e\uff0c\u7528 2% BSA \u5728\u5ba4\u6e29\u4e0b\u5c01\u95ed\u5207\u7247 1 \u5c0f\u65f6\u3002\u5c06\u5207\u7247\u5728 4\u00b0C \u4e0b\u7528 CD45\uff08Bio-RAD\uff0cMCA 1258GT\uff0c1:100 \u7a00\u91ca\uff09\u5904\u7406\u8fc7\u591c\u3002\u7528PBS\u6d17\u6da4\u540e\uff0c\u7528\u6297\u5154IgG-HRP\uff08Santa Cruz Biotechnology\uff0cDallas\uff0cTX\uff0cUSA\uff0cSC-2357\uff0c1:100\u7a00\u91ca\uff09\u5728\u5ba4\u6e29\u4e0b\u5904\u7406\u7ec6\u80de2\u5c0f\u65f6\uff0c\u5e76\u75283\uff0c\u67d3\u82721\u5206\u949f\uff0c 3'-\u4e8c\u6c28\u57fa\u8054\u82ef\u80fa\u56db\u76d0\u9178\u76d0\uff08Vector Laboratories\uff0c\u4f2f\u6797\u683c\u59c6\uff0c\u52a0\u5229\u798f\u5c3c\u4e9a\u5dde\uff0c\u7f8e\u56fd\uff09\u3002\u5c06\u8f7d\u73bb\u7247\u7528\u82cf\u6728\u7cbe\u67d3\u827230\u79d2\u5e76\u7528\u81ea\u6765\u6c34\u51b2\u6d17\u3002\u7136\u540e\u5c06\u5207\u7247\u76d6\u4e0a\u76d6\u73bb\u7247\u5e76\u7528 Permount\u2122 \u5c01\u7247\u5242\uff08Thermo Fisher Scientific\uff0c\u6c83\u5c14\u745f\u59c6\uff0c\u9a6c\u8428\u8bf8\u585e\u5dde\uff0c\u7f8e\u56fd\uff09\u5c01\u7247\u3002\u4f7f\u7528\u5149\u5b66\u663e\u5fae\u955c\uff08Olympus\uff0cCX43\uff0cOlympus Optical Co.\uff0c\u4e1c\u4eac\uff0c\u65e5\u672c\uff09\u68c0\u67e5\u6240\u6709\u8f7d\u73bb\u7247\u5e76\u62cd\u7167\u3002 ImageJ \u8f6f\u4ef6\uff08\u7248\u672c 1.53\uff0c\u7f8e\u56fd\u56fd\u7acb\u536b\u751f\u7814\u7a76\u9662\uff0c\u8d1d\u585e\u65af\u8fbe\uff0c\u9a6c\u91cc\u5170\u5dde\uff0948<\/a><\/sup> \u7528\u4e8e\u8fdb\u884c\u5b9a\u91cf\u6570\u636e\u5206\u6790\u3002<\/p>\n

ARPE-19 \u7ec6\u80de\u57f9\u517b<\/h3>\n

\u4eba\u89c6\u7f51\u819c\u4e0a\u76ae ARPE-19 \u7ec6\u80de\uff08\u7f8e\u56fd\u5178\u578b\u57f9\u517b\u7269\u4fdd\u85cf\u4e2d\u5fc3\uff0cATCC\uff0c\u9a6c\u7eb3\u8428\u65af\uff0c\u5f17\u5409\u5c3c\u4e9a\u5dde\uff0c\u7f8e\u56fd\uff09\u5728\u8865\u5145\u6709 10% FBS\uff08HyClone Laboratories\uff0cLogan\uff0cUT\uff09\u7684 DMEM\/F-12 \u57f9\u517b\u57fa\uff08Gibco\uff0c\u5361\u5c14\u65af\u5df4\u5fb7\uff0c\u52a0\u5229\u798f\u5c3c\u4e9a\u5dde\uff0c\u7f8e\u56fd\uff09\u4e2d\u57f9\u517b\uff0c\u7f8e\u56fd\uff09\u548c 1% \u9752\u9709\u7d20\/\u94fe\u9709\u7d20\uff08HyClone Laboratories\uff09\u3002\u7ec6\u80de\u63a5\u79cd\u5bc6\u5ea6\u4e3a 2 \u00d7 105<\/sup> 6 \u5b54\u677f\u4e2d\u7684\u6bcf\u5b54\u5e76\u57f9\u517b\u4ee5\u8bf1\u5bfc\u708e\u75c7\u548c\u5185\u8d28\u53cd\u5e94\u3002<\/p>\n

\u7ec6\u80de\u6d3b\u529b<\/h3>\n

\u7684\u6548\u679c \u9ec4\u6bdb\u866b<\/i> \u5982\u4e4b\u524d\u7684\u7814\u7a76\u6240\u8ff0\uff0c\u901a\u8fc7 MTT \u6d4b\u5b9a\u8bc4\u4f30\u5bf9\u7ec6\u80de\u6d3b\u529b\u7684\u5f71\u54cd49<\/a><\/sup> \u4f46\u7565\u6709\u4fee\u6539\u3002\u5c06\u7ec6\u80de\u4ee5 1.5 \u00d7 10 \u7684\u5bc6\u5ea6\u63a5\u79cd\u4e8e 96 \u5b54\u57f9\u517b\u677f\u4e2d4<\/sup> \u7ec6\u80de\/\u5b54\u5e76\u5b75\u80b2 24 \u5c0f\u65f6\u3002\u7136\u540e\u7528\u4e0d\u540c\u6d53\u5ea6\u7684 \u9ec4\u6bdb\u866b<\/i> \u5e76\u518d\u5b75\u80b224\u5c0f\u65f6\u3002\u6bcf\u5b54\u4e0a\u6e05\u6db2\u66f4\u6362\u4e3a\u542b\u67090.5 mg\/mL MTT\u7684\u65b0\u9c9c\u751f\u957f\u57f9\u517b\u57fa\uff0c\u5e76\u5b75\u80b21 h\u3002\u9664\u53bb\u4e0a\u6e05\u6db2\u5e76\u5c06\u751f\u6210\u7684\u7532\u81dc\u6c89\u79ef\u7269\u6eb6\u89e3\u5728200\u03bcL\u4e8c\u7532\u57fa\u4e9a\u781c\u4e2d\u3002\u5728 570 nm \u5904\u6d4b\u91cf\u5438\u5149\u5ea6\u3002\u4e0e\u9634\u6027\u5bf9\u7167\u7ec4\u76f8\u6bd4\uff0c\u7ec6\u80de\u6d3b\u529b\u76f8\u5bf9\u8ba1\u7b97\u4e3a\u767e\u5206\u6bd4\u3002<\/p>\n

\u86cb\u767d\u8d28\u5370\u8ff9\u5206\u6790<\/h3>\n

\u8fdb\u884c\u86cb\u767d\u8d28\u5370\u8ff9\u4ee5\u786e\u5b9a\u6297\u708e\u4f5c\u7528\u80cc\u540e\u7684\u7ec6\u80de\u4fe1\u53f7\u901a\u8def \u9ec4\u6bdb\u866b<\/i> \u5982\u5148\u524d\u7814\u7a76\u6240\u8ff049<\/a><\/sup>\uff0c\u4f46\u7a0d\u4f5c\u4fee\u6539\u3002\u5c06\u6c47\u5408\u7684 ARPE-19 \u7ec6\u80de\u5728\u542b\u6709 1% FBS \u548c 1% \u6297\u751f\u7d20\u7684 DMEM:F12 \u4e2d\u9965\u997f 24 \u5c0f\u65f6\u3002\u7136\u540e\u7528\u5904\u7406\u7ec6\u80de \u9ec4\u6bdb\u866b<\/i> \uff0850 \u548c 100 \u03bcg\/mL\uff092 \u5c0f\u65f6\uff0c\u7136\u540e\u7528 20 ng\/mL TNF-\u03b1 \u523a\u6fc0 15 \u5206\u949f\u3002\u7528\u51b0\u51b7\u7684 DPBS \u51b2\u6d17\u7ec6\u80de 3 \u6b21\uff0c\u7136\u540e\u5728\u8865\u5145\u6709\u86cb\u767d\u9176\u548c\u78f7\u9178\u9176\u6291\u5236\u5242\u6df7\u5408\u7269\u7684\u86cb\u767d\u8d28\u63d0\u53d6\u7f13\u51b2\u6db2\uff08iNtRON Biotechnology\uff0c\u4eac\u757f\u9053\uff0c\u97e9\u56fd\uff09\u4e2d\u88c2\u89e3\u3002\u901a\u8fc7\u79bb\u5fc3\uff0814,000\u00d7G<\/i>\uff0c30 \u5206\u949f\uff0c4 \u00b0C\uff09\uff0c\u5e76\u4f7f\u7528 DCTM \u86cb\u767d\u8d28\u6d4b\u5b9a\u8bd5\u5242\u76d2\uff08Bio-Rad\uff0cHercules\uff0cCA\uff0cUSA\uff09\u4f30\u8ba1\u5176\u86cb\u767d\u8d28\u6d53\u5ea6\u3002\u901a\u8fc7 SDS-PAGE \u5206\u79bb\u7b49\u91cf (15\u201325 \u03bcg) \u7684\u7ec6\u80de\u86cb\u767d\u5e76\u8f6c\u79fb\u5230 PVDF \u819c\u4e0a\u3002\u5c06\u819c\u4e0e 5% \u725b\u8840\u6e05\u767d\u86cb\u767d\u6216\u8131\u8102\u5976\u4e00\u8d77\u5b75\u80b2\uff0c\u7136\u540e\u4e0e 1:2000 \u7a00\u91ca\u7684\u4e00\u6297\uff08\u6297 phopho-ERK\u3001\u6297 -ERK\u3001\u6297 - \u78f7\u9178 -JNK\u3001\u6297 -JNK\u3001\u6297 - \u78f7\u9178 -p38\uff09\u4e00\u8d77\u5b75\u80b2\u3001\u6297 p38\u3001\u6297\u78f7\u9178 AKT\u3001\u6297 AKT\u3001\u6297\u78f7\u9178 I\u03baB-\u03b1\u3001\u6297 I\u03baB-\u03b1\u3001\u6297\u78f7\u9178 p65\u3001\u6297 p65\u3001\u6297 \u03b1 \u5fae\u7ba1\u86cb\u767d\u548c\u6297 \u03b2 \u4f5c\u7528\uff09\u5728 4\u00b0C \u548c 12 \u5c0f\u65f6\u3002\u7136\u540e\u7528 TBS-T \u51b2\u6d17\u819c\uff0c\u5e76\u7528 1:10,000 \u7a00\u91ca\u5ea6\u7684\u6297\u5154 HRP \u6807\u8bb0\u7684\u4e8c\u6297\u5728\u5ba4\u6e29\u4e0b\u5904\u7406 1 \u5c0f\u65f6\u3002\u4f7f\u7528 SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Fisher Scientific) \u68c0\u6d4b\u86cb\u767d\u8d28\u6761\u5e26\uff0c\u5e76\u5728 iBright CL1000 \u51dd\u80f6\u8bb0\u5f55\u7cfb\u7edf (Thermo Fisher Scientific) \u4e2d\u53ef\u89c6\u5316\u3002\u4f7f\u7528Image J\u8f6f\u4ef6\uff08\u7248\u672c1.53\uff0cNIH\uff09\u6d4b\u5b9a\u6bcf\u4e2a\u86cb\u767d\u8d28\u5370\u8ff9\u7684\u5bc6\u5ea648<\/a><\/sup>.<\/p>\n

\u603b PUFA \u63d0\u53d6\u91cf \u9ec4\u6bdb\u866b<\/i><\/h3>\n

\u603b PUFA \u63d0\u53d6\u81ea \u9ec4\u6bdb\u866b<\/i> \u4f7f\u7528\u798f\u5c14\u5947\u6280\u672f50<\/a><\/sup>\u3002\u7b80\u800c\u8a00\u4e4b\uff0c\u5c06 100 mg \u6837\u54c1\u653e\u5165\u542b\u6709 5 mL CH \u7684\u5c0f\u74f6\u4e2d3<\/sub>Cl:MeOH (2:1, v\/v) \u6eb6\u6db2\u5e76\u8d85\u58f0\u5904\u7406 15 \u5206\u949f\u3002\u901a\u8fc7\u4ee5 10,000 rpm \u79bb\u5fc3 5 \u5206\u949f\u4f7f\u60ac\u6d6e\u6db2\u6f84\u6e05\u3002\u94a02<\/sub>\u6240\u4ee54<\/sub> \u5c06\u5176\u6dfb\u52a0\u5230\u4e0a\u6e05\u6db2\u4e2d\uff0c\u7136\u540e\u8fc7\u6ee4\u4ee5\u9664\u53bb\u4efb\u4f55\u5269\u4f59\u7684\u6c34\u3002\u8be5\u6b65\u9aa4\u518d\u91cd\u590d\u4e24\u6b21\u3002\u7136\u540e\u5728\u6c2e\u6c14\u4e0b\u6d53\u7f29\u6eb6\u6db2\u4ee5\u83b7\u5f97\u7eaf\u7684\u85fb\u8102\u3002<\/p>\n

\u8102\u80aa\u9178\u7532\u57fa\u5316<\/h3>\n

\u8fdb\u884c\u8102\u80aa\u9178\u7532\u57fa\u5316\u4ee5\u5236\u5907\u7528\u4e8e\u6c14\u76f8\u8272\u8c31\uff08GC\uff09\u5206\u6790\u7684\u8102\u8d28\u6837\u54c1\u3002\u5c06\u63d0\u53d6\u7684\u7269\u8d28\u6dfb\u52a0\u5230 10 mL \u5c0f\u74f6\u4e2d\uff0c\u7136\u540e\u6dfb\u52a0 1 mL 0.5 N NaOH:MeOH \u6df7\u5408\u7269\u3002\u8be5\u6eb6\u6db2\u542b\u6709 400 \u03bcg\/mL \u7684\u5185\u94fe\uff08\u620a\u9178\uff09\u3002\u5c06\u6df7\u5408\u7269\u6447\u52a8 30 s\uff0c\u7136\u540e\u5728 80 \u00b0C \u4e0b\u5b75\u80b2 20 \u5206\u949f\u3002\u7136\u540e\u5c06\u6df7\u5408\u7269\u5728\u5ba4\u6e29\u4e0b\u51b7\u53745\u5206\u949f\uff0c\u7136\u540e\u52a0\u51652mL\u4e09\u6c1f\u5316\u787c:MeOH\uff0c\u7136\u540e\u6405\u62cc30\u79d2\u3002\u5c06\u6df7\u5408\u7269\u5728 80\u00b0C \u4e0b\u5b75\u80b2 20 \u5206\u949f\uff0c\u7136\u540e\u51b7\u5374 10 \u5206\u949f\u3002\u6dfb\u52a0 2 mL \u4f53\u79ef\u7684\u8fc7\u9971\u548c NaCl \u4ee5\u8bf1\u5bfc\u76f8\u5206\u79bb\u548c\u8102\u80aa\u9178\u6536\u96c6\u3002\u63a5\u4e0b\u6765\uff0c1.5 mL n<\/i>\u6dfb\u52a0\u6b63\u5df1\u70f7\u5e76\u5c06\u6df7\u5408\u7269\u6da1\u65cb30\u79d2\uff0c\u7136\u540e\u5728\u5ba4\u6e29\u4e0b\u51b7\u537420\u5206\u949f\u3002\u5206\u79bb\u5404\u5c42\u5e76\u4ece\u6df7\u5408\u7269\u4e2d\u9664\u53bb\u6db2\u4f53\u4e0a\u6e05\u6db2\u5e76\u4f7f\u7528\u9971\u548cNa2<\/sub>\u6240\u4ee54<\/sub> \u7b5b\u9009\u3002<\/p>\n

\u8102\u80aa\u9178\u7684\u9274\u5b9a<\/h3>\n

\u4f7f\u7528\u914d\u5907\u706b\u7130\u79bb\u5b50\u5316\u68c0\u6d4b\u5668\u7684 GC\uff08FID\uff1bAgilent 7890A\uff0cAgilent Technologies\uff0cWilmington\uff0cDE\uff0cUSA\uff09\u5bf9\u8102\u80aa\u9178\u7532\u916f (FAME) \u8fdb\u884c\u68c0\u6d4b\u3002\u5c06\u6837\u54c1 (1 \u00b5L) \u6ce8\u5165 GC (J&W 112-88A7, Agilent Technologies, Wilmington, DE, USA) \u7684 HP-88 \u8272\u8c31\u67f1\uff08100 m \u00d7 250 \u00b5m ID\uff0c0.2 \u00b5m \u8584\u819c\uff09\u3002 GC \u5206\u6790\u5982\u4e0b\u8fdb\u884c\uff1a\u6837\u54c1\u6700\u521d\u5728 140 \u00b0C \u4e0b\u4fdd\u6301 5 \u5206\u949f\uff0c\u7136\u540e\u4ee5 4 \u00b0C\/min \u5347\u6e29\u81f3 240 \u00b0C\uff0c\u6301\u7eed 15 \u5206\u949f\u3002\u67f1\u7684\u6d41\u901f\u56fa\u5b9a\u4e3a1 mL\/min\u3002\u91c7\u7528\u6c2e\u6c14\u4f5c\u4e3a\u5206\u6790\u8f7d\u6c14\uff0cFID\u8bbe\u5b9a\u4e3a280\u2103\uff0c\u5206\u6d41\u6bd4\u4e3a30:1\u3002 FAME \u662f\u901a\u8fc7\u4e0e\u5df2\u77e5\u6807\u51c6\u8fdb\u884c\u6bd4\u8f83\u6765\u8bc6\u522b\u7684\u3002\u6bcf\u4e2a\u6837\u54c1\u5747\u901a\u8fc7 GC \u5206\u6790\u4e00\u5f0f\u4e09\u4efd\u8fdb\u884c\u6d4b\u91cf\u3002<\/p>\n

\u7edf\u8ba1\u5206\u6790<\/h3>\n

\u6240\u6709\u6570\u636e\u5747\u4f7f\u7528\u5355\u5411\u65b9\u5dee\u5206\u6790\u548c Tukey \u4e8b\u540e\u5206\u6790\u901a\u8fc7 SPSS 18.0 \u7248\uff08IBM Co.\uff0cArmonk\uff0cNY\uff0cUSA\uff09\u8fdb\u884c\u7edf\u8ba1\u5206\u6790\u3002\u6bcf\u4e2a\u5b9e\u9a8c\u81f3\u5c11\u8fdb\u884c\u4e09\u6b21\u72ec\u7acb\u91cd\u590d\u3002<\/p>\n<\/div>\n

[\u5e7f\u544a_2]<\/p>\n

\u6765\u6e90\u94fe\u63a5 <\/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\/zh\/wp-json\/wp\/v2\/posts\/4903","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/directcm.com\/zh\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/directcm.com\/zh\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/directcm.com\/zh\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/directcm.com\/zh\/wp-json\/wp\/v2\/comments?post=4903"}],"version-history":[{"count":1,"href":"https:\/\/directcm.com\/zh\/wp-json\/wp\/v2\/posts\/4903\/revisions"}],"predecessor-version":[{"id":5393,"href":"https:\/\/directcm.com\/zh\/wp-json\/wp\/v2\/posts\/4903\/revisions\/5393"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/directcm.com\/zh\/wp-json\/wp\/v2\/media\/5363"}],"wp:attachment":[{"href":"https:\/\/directcm.com\/zh\/wp-json\/wp\/v2\/media?parent=4903"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/directcm.com\/zh\/wp-json\/wp\/v2\/categories?post=4903"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/directcm.com\/zh\/wp-json\/wp\/v2\/tags?post=4903"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}