中华心血管病杂志
中華心血管病雜誌
중화심혈관병잡지
Chinese Journal of Cardiology
2013年
4期
320-326
,共7页
鲍宏刚%张卫泽%马凌%李涛%王菲%陈永清
鮑宏剛%張衛澤%馬凌%李濤%王菲%陳永清
포굉강%장위택%마릉%리도%왕비%진영청
心房%纤维化%谷氨酰胺%HSP70热休克蛋白质类%连接蛋白43
心房%纖維化%穀氨酰胺%HSP70熱休剋蛋白質類%連接蛋白43
심방%섬유화%곡안선알%HSP70열휴극단백질류%련접단백43
Heart atria%Fibrosis%Glutamine%HSP70 heat-shock proteins%Connexin 43
目的 探讨谷氨酰胺(glutamine,Gln)诱导热休克蛋白70高表达对大鼠心房肌纤维化及缝隙连接蛋白43(connexin43,Cx43)重构的影响及其机制.方法 40只SPF级雄性SD大鼠,体质量(180±20)g,随机数字表法随机分为对照组、二甲基亚砜(DMSO)组、盐酸异丙肾上腺素(isoprenaline,ISO)5 mg· kg-1·d-1组(模型组)、ISO5 mg· kg-1·d-1+内氨酰谷氨酰胺(Ala-Gln)0.75 mg·kg-1·d-1组(干预组)和ISO5 mg· kg-1·d-1+槲皮素(quercetin,QUE) 100 mg· kg-1·d-1 +Ala-Gln0.75 mg· kg-1· d-1+DMSO组(QUE组),每组8只大鼠.每组每天1次给予相应试剂,7d后处死大鼠.放射免疫法检测大鼠心房肌组织中血管紧张素Ⅱ(Ang Ⅱ)含量；苏木素伊红(HE)染色法观察大鼠心房纤维化情况；Masson染色法计算心房肌胶原容积分数；免疫组织化学SP法观察并半定量测定大鼠心房肌组织中热休克蛋白70(heat shock protein70,Hsp70)、磷酸化c-Jun氨基末端激酶1/2/3(p-JNK1/2/3)、c-Jun及Cx43的含量.结果 DMSO组大鼠心房肌组织中 AngⅡ含量[(71.47±11.49) pg/L]与对照组[(68.51±10.76) pg/L]比较差异无统计学意义(P＞0.05),而模型组[(211.25±49.49) pg/L]、干预组[(185.32±54.85) ng/L]和QUE组[(189.90 ±42.12) pg/L]则均显著高于对照组(P均＜0.01) HE、Masson染色结果示对照组和DMSO组大鼠心房肌纤维化不明显[心肌胶原容积分数分别为(6.842±1.674)％ 和(7.108±1.343)％],模型组和QUE组则有明显的纤维化[心肌胶原容积分数分别为(29.485±9.966)％和(25.06 ±8.581)％],而干预组纤维化程度[心肌胶原容积分数为(7.861±1.867)％]较模型组和QUE组心房肌纤维化程度弱(心肌胶原容积分数比较P均＜0.01).对照组、DMSO 组、模型组和QUE组心肌组织 Hsp70含量比较差异均无统计学意义(分别为0.160±0.023、0.163±0.022、0.166±0.028和0.168±0.027,P均＞0.05),而干预组(0.215 ±0.018)则显著高于上述各组(P均＜0.01).DMSO组大鼠心房肌组织中p-JNK1/2/3和c-Jun含量(分别为0.154 ±0.021和0.164±0.024)与对照组(分别为0.151 ±0.016和0.163±0.022)比较差异均无统计学意义(P均＞0.05),而模型组(分别为0.202±0.025和0.254±0.044)、QUE组(分别为0.196±0.024和0.251±0.027)均显著高于对照组和DMSO组(P均＜0.01),而干预组(分别为0.160±0.025 和0.168±0.024)与对照组和DMSO组相比差异均无统计学意义(P均＞0.05),且显著低于模型组和QUE组(P均＜0.01). DMSO组大鼠心房肌组织中 Cx43含量与对照组比较差异无统计学意义(0.220±0.032比0.231 ±0.035,P＞0.05)且呈线性规律分布,而模型组(0.163 ±0.013)和 QUE组(0.165 ±0.024)则显著少于对照组(P均＜0.01)且分布无规律性,侧面分布相对增多,干预组(0.216 ±0.026)则较模型组、QUE组多(P均＜0.01)且分布较规律.结论 Gln诱导Hsp70高表达可减轻ISO诱发的大鼠心房肌纤维化程度和Cx43重构,机制可能与Hsp70下调p-JNK1/2/3和c-Jun表达、抑制JNK信号通路激活有关.
目的 探討穀氨酰胺(glutamine,Gln)誘導熱休剋蛋白70高錶達對大鼠心房肌纖維化及縫隙連接蛋白43(connexin43,Cx43)重構的影響及其機製.方法 40隻SPF級雄性SD大鼠,體質量(180±20)g,隨機數字錶法隨機分為對照組、二甲基亞砜(DMSO)組、鹽痠異丙腎上腺素(isoprenaline,ISO)5 mg· kg-1·d-1組(模型組)、ISO5 mg· kg-1·d-1+內氨酰穀氨酰胺(Ala-Gln)0.75 mg·kg-1·d-1組(榦預組)和ISO5 mg· kg-1·d-1+槲皮素(quercetin,QUE) 100 mg· kg-1·d-1 +Ala-Gln0.75 mg· kg-1· d-1+DMSO組(QUE組),每組8隻大鼠.每組每天1次給予相應試劑,7d後處死大鼠.放射免疫法檢測大鼠心房肌組織中血管緊張素Ⅱ(Ang Ⅱ)含量；囌木素伊紅(HE)染色法觀察大鼠心房纖維化情況；Masson染色法計算心房肌膠原容積分數；免疫組織化學SP法觀察併半定量測定大鼠心房肌組織中熱休剋蛋白70(heat shock protein70,Hsp70)、燐痠化c-Jun氨基末耑激酶1/2/3(p-JNK1/2/3)、c-Jun及Cx43的含量.結果 DMSO組大鼠心房肌組織中 AngⅡ含量[(71.47±11.49) pg/L]與對照組[(68.51±10.76) pg/L]比較差異無統計學意義(P＞0.05),而模型組[(211.25±49.49) pg/L]、榦預組[(185.32±54.85) ng/L]和QUE組[(189.90 ±42.12) pg/L]則均顯著高于對照組(P均＜0.01) HE、Masson染色結果示對照組和DMSO組大鼠心房肌纖維化不明顯[心肌膠原容積分數分彆為(6.842±1.674)％ 和(7.108±1.343)％],模型組和QUE組則有明顯的纖維化[心肌膠原容積分數分彆為(29.485±9.966)％和(25.06 ±8.581)％],而榦預組纖維化程度[心肌膠原容積分數為(7.861±1.867)％]較模型組和QUE組心房肌纖維化程度弱(心肌膠原容積分數比較P均＜0.01).對照組、DMSO 組、模型組和QUE組心肌組織 Hsp70含量比較差異均無統計學意義(分彆為0.160±0.023、0.163±0.022、0.166±0.028和0.168±0.027,P均＞0.05),而榦預組(0.215 ±0.018)則顯著高于上述各組(P均＜0.01).DMSO組大鼠心房肌組織中p-JNK1/2/3和c-Jun含量(分彆為0.154 ±0.021和0.164±0.024)與對照組(分彆為0.151 ±0.016和0.163±0.022)比較差異均無統計學意義(P均＞0.05),而模型組(分彆為0.202±0.025和0.254±0.044)、QUE組(分彆為0.196±0.024和0.251±0.027)均顯著高于對照組和DMSO組(P均＜0.01),而榦預組(分彆為0.160±0.025 和0.168±0.024)與對照組和DMSO組相比差異均無統計學意義(P均＞0.05),且顯著低于模型組和QUE組(P均＜0.01). DMSO組大鼠心房肌組織中 Cx43含量與對照組比較差異無統計學意義(0.220±0.032比0.231 ±0.035,P＞0.05)且呈線性規律分佈,而模型組(0.163 ±0.013)和 QUE組(0.165 ±0.024)則顯著少于對照組(P均＜0.01)且分佈無規律性,側麵分佈相對增多,榦預組(0.216 ±0.026)則較模型組、QUE組多(P均＜0.01)且分佈較規律.結論 Gln誘導Hsp70高錶達可減輕ISO誘髮的大鼠心房肌纖維化程度和Cx43重構,機製可能與Hsp70下調p-JNK1/2/3和c-Jun錶達、抑製JNK信號通路激活有關.
목적 탐토곡안선알(glutamine,Gln)유도열휴극단백70고표체대대서심방기섬유화급봉극련접단백43(connexin43,Cx43)중구적영향급기궤제.방법 40지SPF급웅성SD대서,체질량(180±20)g,수궤수자표법수궤분위대조조、이갑기아풍(DMSO)조、염산이병신상선소(isoprenaline,ISO)5 mg· kg-1·d-1조(모형조)、ISO5 mg· kg-1·d-1+내안선곡안선알(Ala-Gln)0.75 mg·kg-1·d-1조(간예조)화ISO5 mg· kg-1·d-1+곡피소(quercetin,QUE) 100 mg· kg-1·d-1 +Ala-Gln0.75 mg· kg-1· d-1+DMSO조(QUE조),매조8지대서.매조매천1차급여상응시제,7d후처사대서.방사면역법검측대서심방기조직중혈관긴장소Ⅱ(Ang Ⅱ)함량；소목소이홍(HE)염색법관찰대서심방섬유화정황；Masson염색법계산심방기효원용적분수；면역조직화학SP법관찰병반정량측정대서심방기조직중열휴극단백70(heat shock protein70,Hsp70)、린산화c-Jun안기말단격매1/2/3(p-JNK1/2/3)、c-Jun급Cx43적함량.결과 DMSO조대서심방기조직중 AngⅡ함량[(71.47±11.49) pg/L]여대조조[(68.51±10.76) pg/L]비교차이무통계학의의(P＞0.05),이모형조[(211.25±49.49) pg/L]、간예조[(185.32±54.85) ng/L]화QUE조[(189.90 ±42.12) pg/L]칙균현저고우대조조(P균＜0.01) HE、Masson염색결과시대조조화DMSO조대서심방기섬유화불명현[심기효원용적분수분별위(6.842±1.674)％ 화(7.108±1.343)％],모형조화QUE조칙유명현적섬유화[심기효원용적분수분별위(29.485±9.966)％화(25.06 ±8.581)％],이간예조섬유화정도[심기효원용적분수위(7.861±1.867)％]교모형조화QUE조심방기섬유화정도약(심기효원용적분수비교P균＜0.01).대조조、DMSO 조、모형조화QUE조심기조직 Hsp70함량비교차이균무통계학의의(분별위0.160±0.023、0.163±0.022、0.166±0.028화0.168±0.027,P균＞0.05),이간예조(0.215 ±0.018)칙현저고우상술각조(P균＜0.01).DMSO조대서심방기조직중p-JNK1/2/3화c-Jun함량(분별위0.154 ±0.021화0.164±0.024)여대조조(분별위0.151 ±0.016화0.163±0.022)비교차이균무통계학의의(P균＞0.05),이모형조(분별위0.202±0.025화0.254±0.044)、QUE조(분별위0.196±0.024화0.251±0.027)균현저고우대조조화DMSO조(P균＜0.01),이간예조(분별위0.160±0.025 화0.168±0.024)여대조조화DMSO조상비차이균무통계학의의(P균＞0.05),차현저저우모형조화QUE조(P균＜0.01). DMSO조대서심방기조직중 Cx43함량여대조조비교차이무통계학의의(0.220±0.032비0.231 ±0.035,P＞0.05)차정선성규률분포,이모형조(0.163 ±0.013)화 QUE조(0.165 ±0.024)칙현저소우대조조(P균＜0.01)차분포무규률성,측면분포상대증다,간예조(0.216 ±0.026)칙교모형조、QUE조다(P균＜0.01)차분포교규률.결론 Gln유도Hsp70고표체가감경ISO유발적대서심방기섬유화정도화Cx43중구,궤제가능여Hsp70하조p-JNK1/2/3화c-Jun표체、억제JNK신호통로격활유관.
Objective To explore the effects of glutamine (Gln) induced heat shock protein 70 (Hsp70) overexpression on atrial fibrosis and connexin43 remodeling in isoprenaline(ISO) treated rats and related mechanisms.Methods Forty male SD rats were randomly divided into five groups (n =8 each group):control group,DMSO group,ISO5 mg · kg-1 · d-1 group (Fibrosis group),ISO5 mg· kg-1 ·d-1 +Ala-Gln0.75 mg· kg-1 ·d-1 group(Intervention group) and ISO 5 mg·kg-1 ·d-1 +QUE 100 mg·kg-1 ·d-1 +Ala-Gln0.75 mg· kg-1 · d-1 +DMSO group(QUE group).Rats were killed after7 d.The Ang Ⅱ expression in myocardial tissue was detected by radioimmunoassay; myocardial fibrosis was observed by HE staining.Collagen volume fractions were quantified by Masson staining and as the indicators of atrial fibrosis.The expressions of Hsp70,p-JNK1/2/3,c-Jun and Cx43 were determined with immunohistochemical method.Results Ang Ⅱ content was similar between the control group[(68.51 ± 10.76) pg/L] and DMSO [(71.47±11.49) pg/L] group (P＞ 0.05),and significantly increased in fibrosis group[(211.25 ±49.49)pg/L],intervention group[(185.32 ± 54.85) pg/L] and QUE[(189.90 ± 42.12) pg/L] group (P ＜ 0.01 vs.control group).Atrial fibrosis was significantly higher in the fibrosis group [(29.485 ± 9.966)％] and QUE group[(25.060 ±8.581)％] but not in the intervention group[(7.861 ± 1.867)％]compared to control group [(6.842 ± 1.674) ％] and DMSO group [(7.108 ± 1.343) ％].The expression of Hsp70 was similar among the control group (0.160 ± 0.023),DMSO group (0.163 ± 0.022),fibrosis group(0.166 ±0.028) and QUE(0.168 ±0.027) group (P ＞0.05) while significantly upregulated in the intervention group (0.215 ± 0.018) (P ＜ 0.01 vs.control group).The expressions of p-JNK1/2/3 and c-Jun were similar between control group(0.151 ±0.016;0.163 ±0.022) and DMSO group(0.154 ±0.021 ;0.164 ± 0.024) (P ＞ 0.05),while significantly upregulated in fibrosis group (0.202 ± 0.025 ; 0.254 ± 0.044) and QU E group (0.196 ± 0.024 ; 0.251 ± 0.027) (P ＜ 0.01 vs.control group) but not in intervention group(0.160 ±0.025 ; 0.168 ±0.024) were not changed obviously(P ＞0.05 vs.control group).The content of Cx43 was similar between control group and DMSO group(0.231 ± 0.035 vs.0.220 ± 0.032,P ＞ 0.05),and was linearly distributed in intercalated disc of the cardiomyocytes,however,the content of Cx43 was significantly reduced(P ＜ 0.01)and the Cx43 distribution was disordered in fibrosis group(0.163 ± 0.013) and QUE group(0.165 ± 0.024),while these changes were not found in intervention group.Conclusion Glutamine could reduce the atrial fibrosis and Cx43 remodeling in isoprenaline-treated rats by up-regulating Hsp70 and inhibiting JNK signaling pathway activation through down-regulating p-JNK1/2/3 and c-Jun expression.
