ѧλÂÛÎÄ4 - ͼÎÄ ÁªÏµ¿Í·þ

·¢²¼Ê±¼ä : ÐÇÆÚÈÕ ÎÄÕÂѧλÂÛÎÄ4 - ͼÎĸüÐÂÍê±Ï¿ªÊ¼ÔĶÁ903cf73ebb68a98271fefaf0

ÐìÖÝҽѧԺ˶ʿÂÛÎÄ

To verify TLR4 mRNA expression changes in the HUVEC, RT-PCR and quantitative real time PCR were performed. The TLR4 mRNA expression levels were significantly increased in the VIN-treated HUVEC (P<0.05), compared with vehicle-treated group. The effect of VIN reached a maximum at the time after exposure 1h; We performed western blot for the relative protein expression of TLR4 in HUVECs to assess the effect after treatment with VIN. The expression of TLR4 protein in all VIN-treated groups was increased significantly compared with vehicle-treated group(P<0.05); NF-¦ÊB activation as determined by NF-¦ÊB phosphorylation was detected at different time and was no longer detectable at later time points. As expected, nuclear translocation of p65 occurred in HUVEC treated with VIN, and NF-¦ÊB activation occurred within a very narrow window in HUVEC during the course of vascular endothelial injury.

Conclusions 1. Pathohistological changes of the model group were observed, such as loss of venous endothelial cells, inflammatory cell infiltration and thrombus. 2. mouse and cell experiments indicates that TLR4 expression is up-regulated in vascular endothelial injury that induced by Vinorelbine. 3. cell experiment indicates that TLR4 mRNA and protein are up-regulated by Vinorelbine in HUVEC. 4. cell experiment indicates that nuclear translocation of p65 occurred in HUVEC treated with VIN. 5. Tegether, our results suggest that TLR4 may play an important role in the development of vascular endothelial injury that induced by Vinorelbine.

Key words Toll-like receptor; Vinorelbine; Phlebitis; HUVEC; Mechanism

6

ÐìÖÝҽѧԺ˶ʿÂÛÎÄ

Ç° ÑÔ

Ëæ׏¤Òµ»¯µÄ½øÕ¹¡¢»·¾³ÎÛȾµÄ¼Ó¾ç£¬ÈËÃǵÄÉú»î½Ú×àÔ½À´Ô½¿ì£¬²»Á¼Éú»îÏ°¹ß¼°²»Á¼ÒûʳµÄÓ°Ï죬Ö×ÁöµÄ·¢²¡ÂÊÒ²ÖðÄêÔö¸ß£¬ÎÞÂÛÔÚ·¢´ï¹ú¼Ò»¹ÊÇ·¢Õ¹Öйú¼Ò£¬¶ñÐÔÖ×ÁöµÄΣº¦¶¼²»ÈݺöÊÓ¡£¾²ÂöÊä×¢¿¹Ö×ÁöÒ©ÎïÊÇÖÎÁƶñÐÔÖ×ÁöµÄÖ÷Òª·½·¨Ö®Ò»£¬µ«ÍùÍùÒýÆðÑÏÖصĶ¾¸±·´Ó¦£¬³ý¶ÔÔìѪϵͳ¼°È«ÉíÖØÒªÔàÆ÷¾ßÓж¾ÐÔÒÔÍ⣬¶ÔѪ¹ÜÖÜΧ×éÖ¯»¹¿ÉÒýÆðÑÏÖصÄÑ×ÐԴ̼¤·´Ó¦¡£¾Ö²¿×éÖ¯±íÏֺ졢Öס¢ÈÈ¡¢Í´µÈÑ×Ö¢·´Ó¦£¬ÓÐʱÑؾ²Âö×ßÏò³öÏÖÌõË÷×´¸Ä±ä£¬ÑÏÖØÕß¾Ö²¿×éÖ¯ÃÓÀᢻµËÀ¡£

Ö×Áö»¼Õß³¤ÆÚ·Å¡¢»¯ÁÆ£¬»úÌåÃâÒßÁ¦µÍÏÂʱ£¬³£µ¼ÖÂÈ«Éí¸ÐȾ¶øΣ¼°ÉúÃü¡£

³¤´ºÈð±õÊÇÁÙ´²³£¼ûµÄ¿¹Ö×ÁöÒ©ÎÊôÓÚ³¤´º»¨ÉúÎï¼îÀàÒ©ÎËüµÄ¿¹Ö×Áö»îÐÔÖ÷Ҫͨ¹ý¸ÉÈÅ΢¹Üµ°°×¶øÒÖÖÆÖÐÆÚÓÐË¿·ÖÁÑ£¬Ä¿Ç°Ö÷ÒªÓÃÓÚ·ÇСϸ°û·Î°©ÒÔ¼°ÆäËûʵÌåÁöµÈ¡£È»¶øÔÚ³¤´ºÈð±õ¸ø²¡ÈË´øÀ´¿¹Ö×ÁöÁÆЧµÄͬʱ£¬Ëü×÷ΪÖжȷ¢ðå¼ÁµÄÒ»ÖÖ£¬Ò²³£³£ÒýÆð¾Ö²¿¶¾¸±·´Ó¦£¬¾Ö²¿×éÖ¯·¢ºì£¬ÌÛÍ´¡¢Ö×ÕÍÉõÖÁÃÓÀûµËÀ¡£¾Ý±¨µÀ[1]£¬ÆäÖ¾²ÂöÑ׵ķ¢ÉúÂÊ¿É´ï24-33%£¬¹úÄÚѧÕß±¨µÀÆä·¢ÉúÂÊÉõÖÁ¿É´ï87%[2]¡£ËäÈ»ÕâÖÖÒ©ÎïÐÔ¾²ÂöÑ×ÔÚÁÙ´²³£¼û£¬µ«ÊÇÆäÈ·ÇлúÖÆÈÔ²»Ã÷È·£¬Ä¿Ç°ÆÕ±éµÄ¹ÛµãÈÏΪҩÎïÐÔ¾²ÂöÑ×ÊÇÓÉÓÚÒ©Îï¶ÔѪ¹ÜÄÚƤϸ°ûµÄ»¯Ñ§ÐԴ̼¤¶øÒýÆðµÄÎÞ¾úÐÔÑ×Ö¢¡£

Ò©ÎïÐÔ¾²ÂöÑ×µÄÐγÉÒòËØ°üÀ¨Ò©ÎïÖÖÀà¡¢Ò©ÒºµÄζȡ¢Ò©ÒºµÄPHÖµ¡¢Ò©ÒºµÄÉø͸ѹ¡¢Ò©Îï±¾ÉíµÄ¶¾ÐÔ×÷Óá¢Ò©ÎïÒýÆðµÄ±ä̬·´Ó¦ÒÔ¼°Ò©ÒºµÄÊäҺ΢Á£µÈ¡£ÁíÍâÒ©ÎïÐԵľ²ÂöÑ×·¢Éú»¹ÓëÊäÈëÒ©ÒºµÄ¼ÁÁ¿¡¢Å¨¶È¡¢ÊäÈëËٶȡ¢Ê±¼äÒÔ¼°Êä×¢·½·¨µÈÓйØ[2-28]¡£¿¹ÉúËغͿ¹Ö×ÁöÒ©Îïµ¼Öµľ²ÂöÑ׵ķ¢ÉúÂÊ×î¸ß[29-30]£¬¿¹Ö×ÁöÒ©ÎïÖÐÒÔ³¤´ºÈð±õ×îΪÑÏÖØ¡£¾¡¹ÜÁÙ´²¹¤×÷Õß²ÉÈ¡ÁËÐí¶à·À·¶´ëÊ©£¬ÀýÈç¸øÒ©²¿Î»ÈÈ·ó¡¢Àä·óµÄ·½·¨£¬ÒÔ¼°¾Ö²¿¸øÓèÌÇƤÖʼ¤ËØ¡¢·ÇçÞÌ忹Ñ×Ò©¡¢¿¹×é°·Ò©¡¢Ñª¹ÜÀ©ÕżÁ¡¢¾Ö²¿Âé×íÒ©¡¢¿¹ÄýÒ©¡¢ÈÜ˨ҩÉõÖÁ»îѪ»¯ðöÀàÖÐÒ©µÈ£¬µ«Òź¶µÄÊÇÓÉ

7

ÐìÖÝҽѧԺ˶ʿÂÛÎÄ

ÓÚȱ·¦ÏàÓ¦µÄ»ù´¡Ñо¿Ö§³Ö£¬Ê¹µÃÕâЩ´ëÊ©Õë¶ÔÐÔ²»Ç¿£¬ÁÙ´²Ð§¹û²»Ã÷ÏÔ£¬ÕâÖÖÓÉÒ©Îï×ÔÉíÒòËØÒýÆðµÄ¾²ÂöÑײ¢Î´µÃµ½¸ÄÉÆ¡£

½üÄêÀ´Óйؾ²ÂöÑ׵Ļù´¡Ñо¿Ö÷ÒªÊÇÕë¶Ô¿¹ÉúËØÀàÒ©ÎïµÄ¡£Ä¿Ç°¹úÄÚÍâ×ÊÁÏÉÐȱ·¦¶Ô»¯ÁÆÒ©ÎÌرðÊdz¤´º¼îÀ໯ÁÆÒ©ÎïËùÖ¾²ÂöÑ׵Ļù´¡Ñо¿£¬»¯ÁÆÒ©ÎïËùÖ¾²ÂöÑ׵Ļù´¡Ñо¿Ò²½ö¾ÖÏÞÓÚ¶¯ÎﲡÀíѧˮƽ£¬ÉîÈëµÄ»úÖÆÑо¿Ïʼû±¨µÀ£¬Óɴ˿ɼû£¬ÉîÈëÑо¿Ò©Îï¾²ÂöÑ×·¢²¡»úÖƵÄÐèÇó·Ç³£ÆÈÇУ¬ÔÚ´Ë»ù´¡ÉÏÑ°ÕÒÁÙ´²ÉϸüΪÌØÒìºÍÓÐЧµÄÔ¤·À»òÖÎÁÆÒ©Î¶ÔÌá¸ßºÏÀíÓÃҩˮƽ£¬¼õÉÙÒ©Ô´ÐÔ¼²²¡Ë𺦣¬¸ÄÉÆ»¼ÕßµÄÉú»îÖÊÁ¿¾ßÓÐÖØÒªµÄÀíÂÛºÍʵ¼ÊÒâÒå¡£

¹úÍâѧÕßÑо¿[31-32]±íÃ÷Ѫ¹ÜÄÚƤϸ°ûÔÚÒ»¶¨Å¨¶ÈµÄ¸ßϸ°û¶¾ÐԵĿ¹ÉúËصĴ̼¤Ï£¬Ï¸°û¼ä𤸽·Ö×Ó1(intercellular adhesion molecule-1£¬ICAM-1)¡¢E-Ñ¡ÔñËØ(E-selectin)µÈ𤸽·Ö×ӵıí´ïÔö¸ß£¬¶øÕâЩϸ°û¼äÏ໥×÷ÓÃÏà¹ØµÄϸ°û±íÃæ±ê¼ÇÎïµÄ±í´ï£¬¿ÉÄÜÊÇÒ©ÎïÐÔ¾²ÂöÑ×ÖØÒªµÄ·¢Éú»úÖÆ¡£ÄÇôÁÙ´²³£¼ûµÄ³¤´ºÈð±õËùÖµľ²ÂöÑ×ÖÐÊÇ·ñÒ²´æÔÚÕâЩϸ°ûÒò×ӵıí´ï±ä»¯£¿Òà»ò´æÔÚÆäËûÑ×Ö¢Ïà¹Øϸ°ûÒò×ӵıí´ïÄØ£¿

TollÑùÊÜÌå(Toll-like receptor, TLR)ÊÇÒ»Àà½éµ¼ÌìÈ»ÃâÒߵĿçĤÐźŴ«µÝÊÜÌå¼Ò×壬ÔÚϸ°û»î»¯ÐźŵÄתµ¼ÖÐÆðÖØÒª×÷Óã¬ÊÇÁªÏµÌìÈ»ÃâÒßÓë»ñµÃÐÔÃâÒßµÄÇÅÁº[33]¡£±»ÈÏΪÊÇÄ¿Ç°²¸È鶯ÎïΨһ½«Ï¸°ûÍ⿹ԭʶ±ðÐÅÏ¢Ïòϸ°ûÄÚ´«µÝ²¢Òý·¢Ñ×Ö¢·´Ó¦µÄ¹Ø¼ü¿çĤµ°°×[34]¡£

²¸È鶯ÎïÖÐÒÑ·¢ÏÖÖÁÉÙ13ÖÖTLRºÍһϵÁÐTLRÅäÌ壬ÆäÖÐÈËTLRΪTLR1¡«10£¬ËüÃÇ·Ö±ðÔÚ²»Í¬µÄÃâÒßÓ¦´ð»úÖÆÖаçÑݸ÷×ÔµÄÖØÒª½ÇÉ«¡£ÃâÒßϸ°ûͨ¹ýÆä±íÃæµÄTollÑùÊÜÌå¼Ò×å(Toll-like receptors, TLRs)ʶ±ð²¡Ô­Î¢ÉúÎïÆô¶¯ÌìÈ»ÃâÒß·´Ó¦¡£¸÷ÖÖ²»Í¬µÄ²¡Ô­Ì壬°üÀ¨¸ïÀ¼ÑôÐÔ¾ú¡¢¸ïÀ¼ÒõÐÔ¾ú¡¢Õæ¾ú¡¢²¡¶¾ºÍÒÂÔ­Ìå¾ù¿ÉÒÔ±»TLRsʶ±ð¡£ÔÚÌìÈ»ÃâÒß¹ý³ÌÖУ¬TLRs²¶»ñ²¡Ô­ÌåÉÏÏàÓ¦ÅäÌ壬Æô¶¯Ï¸°ûÄÚÐźÅ;¾¶£¬µ¼ÖÂһϵÁÐøµÄ¼¤»î¡¢ºËÒò×Ó-¦ÊB£¨nuclear factor-¦ÊB£¬NF-¦ÊB£©Í¾¾¶»î»¯¼°Ñ×Ö¢Òò×ÓµÄÊÍ·Å¡£

8

ÐìÖÝҽѧԺ˶ʿÂÛÎÄ

TLR4ÔÚ¸ÃÊÜÌå¼Ò×åÖÐ×îÔç±»·¢ÏÖ£¬¶¨Î»ÓÚÈËȾɫÌå9q32-q33ÉÏ£¬×éÖ¯·Ö²¼½Ï¹ã·º£¬¼ûÓÚѪ¹ÜÄÚƤϸ°û¡¢µ¥ºËϸ°û¡¢ÖÐÐÔÁ£Ï¸°û¡¢Ê÷ͻϸ°û¡¢Ð¡³¦ÉÏƤϸ°û¡¢ºôÎüµÀÉÏƤϸ°û¡¢È˳Ýö¸³ÉÏËάϸ°û¡¢ÈË×Ó¹¬¾±Æ½»¬¼¡Ï¸°û¡¢´óÊóÆ¢ºÍÐļ¡Ï¸°ûµÈ£¬Ö÷Òª¹¦ÄÜÊÇ×÷Ϊ֬¶àÌÇ(lipopolysaccharides£¬LPS)µÄÐźÅתµ¼ÊÜÌ壬ÔÚËÞÖ÷µÄ·ÀÓù·´Ó¦Öз¢»ÓÖØÒª×÷ÓÃ[35-36]¡£

TLR4Ö÷Ҫʶ±ð¸ïÀ¼ÒõÐÔ¾ú³É·ÝLPS£¬LPSͨ¹ý¼¤»îËÞÖ÷ϸ°ûĤÉϵÄCD14ºÍTLR4ÊÜÌ壬½øÒ»²½»î»¯NF-§ÜB£¬Ê¹µ¥ºËºÍ¾ÞÊÉϸ°û²úÉúIL-21¡¢6¡¢8ºÍÖ×Áö»µËÀÒò×Ó¦Á(Tumor necrosis factor-¦Á£¬TNF-¦Á)£¬Ç÷»¯Ñ×ÐÔϸ°û½þÈó£¬ÓÕ·¢Ñ×Ö¢£¬´Ì¼¤Ê÷ͻ״ϸ°û³ÉÊ죬Æô¶¯ÏÈÌìºÍºóÌìÃâÒß¡£Ñо¿±íÃ÷£¬TLR4²»½öͨ¹ýʶ±ð¸ïÀ¼ÊÏÒõÐÔ¾úµÄLPS Æô¶¯Ï¸°ûÄÚÐźÅתµ¼£¬Í¬Ê±Ò²Ê¶±ð½áºË·ÖÖ¦¸Ë¾ú¡¢ÑÌÇú¾ú¡¢ÐÂÐÍÒþÇò¾úºÍ°×É«ÄîÖé¾úµÈÆäËû΢ÉúÎïµÄ²¡Ô­Ïà¹Ø·Ö×Óģʽ[37]¡£´óÁ¿Ñо¿ÒÑ֤ʵTLR4ÔÚ°ÜѪ֢¡¢Å§¶¾Ö¢¡¢³¦Ñס¢À»Î²Ñס¢ÒÈÏÙÑ׵ȸÐȾÐÔÑ×Ö¢·´Ó¦Öж¼·¢»ÓÁËÖØÒª×÷Óá£

Ô½À´Ô½¶àµÄÖ¤¾ÝÏÔʾ£¬¼´Ê¹Ã»ÓиÐȾ´æÔÚ£¬ÄÚÔ´ÐÔÅäÌåÒ²¿ÉÒԴ̼¤TLR4²¢Òý·¢ÃâÒß·´Ó¦[38]¡£ÎªÇø±ð΢ÉúÎïÀ´Ô´µÄÍâÔ´ÐÔ·Ö×Ó(ÈçLPSµÈ)£¬½«ÆäÃüÃûΪTLRsµÄÄÚÔ´ÐÔÅäÌ塣Ŀǰ·¢ÏÖTLR4¿ÉÒÔʶ±ð¶àÖÖÄÚÔ´ÐÔÅäÌ壬Èç͸Ã÷ÖÊËá¡¢ÁòËá¸ÎËØ¡¢ÏËάµ°°×ËØÔ­¡¢¸ßǨÒÆÂÊ×åµ°°×1ºÍÈÈÐݿ˵°°×µÈ¡£Ñо¿·¢ÏÖ³ýÁËÍâÔ´ÐÔÅäÌåÒÔÍâ, ÄÚÔ´ÐÔ·Ö×Ó£¬±ÈÈçÈÈÐݿ˵°°×60(HSP60)¡¢ÏËά½áºÏËØ¡¢ÏËάµ°°×Ô­ºÍ͸Ã÷ÖÊÍéµÍ¾ÛÌÇÒ²Äܹ»Í¨¹ýTLR4ÓÕµ¼´ÙÑ×ϸ°ûÒò×ӵIJúÉú[39]¡£ÁíÒ»ÏîÑо¿Ò²±íÃ÷£¬Ä³Ð©´´É˺ÍÊÖÊõµÈÒýÆðµÄÈ«ÉíÑ×Ö¢·´Ó¦×ÛºÏÖ¢ºÍÔ¶Àë×éÖ¯Æ÷¹ÙËðÉËʱ£¬Ñ­»·Öв¢Î´¼ì²âµ½Ï¸¾úºÍÄÚ¶¾ËØ£¬¶øÓ¦ÓÃTLR4×è¶Ï¼Á¿ÉÃ÷ÏÔ¼õÇáÑ×Ö¢·´Ó¦ºÍ×éÖ¯ËðÉË£¬Òò´Ë¿ÉÄܲ»ÊÇÄÚ¶¾ËØ£¬¶øÊÇÄÚÔ´ÐÔÅäÌ弤»îÁËTLR4[40]¡£»úÌåÊܵ½´Ì¼¤Ê±£¬ÌåÄÚijЩÄÚÔ´ÐÔ·Ö×ÓÄÜ´Ó¼¤»îµÄÃâÒßϸ°û¡¢³¦ÉÏƤϸ°ûÖ÷¶¯·ÖÃÚ»òÕß´Ó»µËÀ»òËðÉ˵Äϸ°ûÖỶ¯ÊͷųöÀ´¡£ÕâЩÄÚÔ´ÐÔ·Ö×ӿɱ»TLR4ʶ±ð²¢½áºÏ£¬½øÒ»²½ÓÕµ¼µ¥ºË-¾ÞÊÉϸ°ûϵͳ¼°Ê÷ͻ״ϸ°ûµÄ»î»¯ºÍ³ÉÊ죬´Ù½øÑ×Ö¢Òò×ÓµÄÊÍ·Å[41]¡£ÒÔÉÏÑо¿¾ù±íÃ÷TLR4ÔڷǸÐȾ

9