中华神经医学杂志
中華神經醫學雜誌
중화신경의학잡지
CHINESE JOURNAL OF NEUROMEDICINE
2008年
8期
762-766
,共5页
YANG Chang-chun%张一%王强%李璐%WANG Shi-lei%林其谁%XUAN Hai-xing%周岱
YANG Chang-chun%張一%王彊%李璐%WANG Shi-lei%林其誰%XUAN Hai-xing%週岱
YANG Chang-chun%장일%왕강%리로%WANG Shi-lei%림기수%XUAN Hai-xing%주대
白介素-10%钠氛交换器-1%脑缺血再灌注损伤
白介素-10%鈉氛交換器-1%腦缺血再灌註損傷
백개소-10%납분교환기-1%뇌결혈재관주손상
Interleukin-10%NHE-1%Cerebral ischemia reperfusion injury
目的 观察SA脂质体介导入白介素-10(hIL-10)基因转染对脑缺血再灌注损伤模型大鼠半影区钠氢交换器-1(NHE-1)基因表达的影响,探讨IL-10缺血脑保护的作用机制.方法 成年雄性SD大鼠78只按随机数字表法分为正常对照组(6只)、缺血对照组(24只)、hIL-10基因转染组(24只)、空质粒组(24只).采用Longa法建立大鼠局灶性脑缺血再灌注损伤模型.正常对照组不做任何操作;缺血对照组仅做大脑巾动脉阻塞(MCAO)模型和立体定向操作,不注射任何药物;hIL-10基因转染组和空质粒组在建立MCAO模型后,采用立体定向方式分别将SA脂质体/pcDNA3.1-hIL-10混合物或SA脂质体/pcDNA3.1混合物注射入大鼠侧脑室内.24、72、168 h分别用RT-PCR和ELISA检测其转染效果,TTC染色测定脑梗死体积,采用荧光实时定量PCR技术观察各组大鼠脑组织中NHE-1 mRNA和核转录因子NF-κB mRNA的表达水平.结果 (1)RT-PCR结果 :hIL-10基因转染组人鼠在缺血再灌注72 h时,其皮层和海马中均能检测到hIL-10mRNA的表达,而正常对照组、缺血对照组和空质粒组中则不能检测剑hIL-10 mRNA.ELISA结果 :基因转染后24 h(764.63±87.88)脑组织中hIL-10蛋白与正常对照组(81.23±8.61)比较明显升高,72h(1310.21±86.19)较24h更高,但到168 h(541.32±80.77)时已明显下降,但仍高于缺血对照组和空质粒组差异均有统计学意义(P＜0.05).同时hlL-10基因转染组大鼠脑梗死体积明显小于缺血对照组和空质粒组差异均有统计学意义(P＜0.05).(2)正常对照组、缺血对照组、空质粒组和hIL-10基因转染组NF-κBmRNA的表达量分别为1.00±0.33、4.76±0.41、4.58±0.62和2.77±0.43.与正常对照组相比,其它三组NF-κB mRNA的表达量均升高,差异有统计学意义(p＜0.01).但hIL-10基因转染组的升高水平低于缺血对照组和空质粒组,差异有统计学意义(p＜0.01).(3)正常对照组、缺血对照组、空质粒组和hIL-10基囚转染组NHE-1 mRNA的表达量分别为1.00±0.22、4.16±0.48、3.97±0.51和2.82±0.47.与正常对照组相比,后三组NHE-1 mRNA的表达量均升高,差异有统计学意义(P＜0.01),其中hIL-10基因转染组的升高水平低于其它两组差异有统计学意义(P＜0.01).结论 hIL-10基因转染可能通过抑制脑缺血再灌注引起的NHE-1基因表达的增加而发挥缺血脑保护作用.
目的 觀察SA脂質體介導入白介素-10(hIL-10)基因轉染對腦缺血再灌註損傷模型大鼠半影區鈉氫交換器-1(NHE-1)基因錶達的影響,探討IL-10缺血腦保護的作用機製.方法 成年雄性SD大鼠78隻按隨機數字錶法分為正常對照組(6隻)、缺血對照組(24隻)、hIL-10基因轉染組(24隻)、空質粒組(24隻).採用Longa法建立大鼠跼竈性腦缺血再灌註損傷模型.正常對照組不做任何操作;缺血對照組僅做大腦巾動脈阻塞(MCAO)模型和立體定嚮操作,不註射任何藥物;hIL-10基因轉染組和空質粒組在建立MCAO模型後,採用立體定嚮方式分彆將SA脂質體/pcDNA3.1-hIL-10混閤物或SA脂質體/pcDNA3.1混閤物註射入大鼠側腦室內.24、72、168 h分彆用RT-PCR和ELISA檢測其轉染效果,TTC染色測定腦梗死體積,採用熒光實時定量PCR技術觀察各組大鼠腦組織中NHE-1 mRNA和覈轉錄因子NF-κB mRNA的錶達水平.結果 (1)RT-PCR結果 :hIL-10基因轉染組人鼠在缺血再灌註72 h時,其皮層和海馬中均能檢測到hIL-10mRNA的錶達,而正常對照組、缺血對照組和空質粒組中則不能檢測劍hIL-10 mRNA.ELISA結果 :基因轉染後24 h(764.63±87.88)腦組織中hIL-10蛋白與正常對照組(81.23±8.61)比較明顯升高,72h(1310.21±86.19)較24h更高,但到168 h(541.32±80.77)時已明顯下降,但仍高于缺血對照組和空質粒組差異均有統計學意義(P＜0.05).同時hlL-10基因轉染組大鼠腦梗死體積明顯小于缺血對照組和空質粒組差異均有統計學意義(P＜0.05).(2)正常對照組、缺血對照組、空質粒組和hIL-10基因轉染組NF-κBmRNA的錶達量分彆為1.00±0.33、4.76±0.41、4.58±0.62和2.77±0.43.與正常對照組相比,其它三組NF-κB mRNA的錶達量均升高,差異有統計學意義(p＜0.01).但hIL-10基因轉染組的升高水平低于缺血對照組和空質粒組,差異有統計學意義(p＜0.01).(3)正常對照組、缺血對照組、空質粒組和hIL-10基囚轉染組NHE-1 mRNA的錶達量分彆為1.00±0.22、4.16±0.48、3.97±0.51和2.82±0.47.與正常對照組相比,後三組NHE-1 mRNA的錶達量均升高,差異有統計學意義(P＜0.01),其中hIL-10基因轉染組的升高水平低于其它兩組差異有統計學意義(P＜0.01).結論 hIL-10基因轉染可能通過抑製腦缺血再灌註引起的NHE-1基因錶達的增加而髮揮缺血腦保護作用.
목적 관찰SA지질체개도입백개소-10(hIL-10)기인전염대뇌결혈재관주손상모형대서반영구납경교환기-1(NHE-1)기인표체적영향,탐토IL-10결혈뇌보호적작용궤제.방법 성년웅성SD대서78지안수궤수자표법분위정상대조조(6지)、결혈대조조(24지)、hIL-10기인전염조(24지)、공질립조(24지).채용Longa법건립대서국조성뇌결혈재관주손상모형.정상대조조불주임하조작;결혈대조조부주대뇌건동맥조새(MCAO)모형화입체정향조작,불주사임하약물;hIL-10기인전염조화공질립조재건립MCAO모형후,채용입체정향방식분별장SA지질체/pcDNA3.1-hIL-10혼합물혹SA지질체/pcDNA3.1혼합물주사입대서측뇌실내.24、72、168 h분별용RT-PCR화ELISA검측기전염효과,TTC염색측정뇌경사체적,채용형광실시정량PCR기술관찰각조대서뇌조직중NHE-1 mRNA화핵전록인자NF-κB mRNA적표체수평.결과 (1)RT-PCR결과 :hIL-10기인전염조인서재결혈재관주72 h시,기피층화해마중균능검측도hIL-10mRNA적표체,이정상대조조、결혈대조조화공질립조중칙불능검측검hIL-10 mRNA.ELISA결과 :기인전염후24 h(764.63±87.88)뇌조직중hIL-10단백여정상대조조(81.23±8.61)비교명현승고,72h(1310.21±86.19)교24h경고,단도168 h(541.32±80.77)시이명현하강,단잉고우결혈대조조화공질립조차이균유통계학의의(P＜0.05).동시hlL-10기인전염조대서뇌경사체적명현소우결혈대조조화공질립조차이균유통계학의의(P＜0.05).(2)정상대조조、결혈대조조、공질립조화hIL-10기인전염조NF-κBmRNA적표체량분별위1.00±0.33、4.76±0.41、4.58±0.62화2.77±0.43.여정상대조조상비,기타삼조NF-κB mRNA적표체량균승고,차이유통계학의의(p＜0.01).단hIL-10기인전염조적승고수평저우결혈대조조화공질립조,차이유통계학의의(p＜0.01).(3)정상대조조、결혈대조조、공질립조화hIL-10기수전염조NHE-1 mRNA적표체량분별위1.00±0.22、4.16±0.48、3.97±0.51화2.82±0.47.여정상대조조상비,후삼조NHE-1 mRNA적표체량균승고,차이유통계학의의(P＜0.01),기중hIL-10기인전염조적승고수평저우기타량조차이유통계학의의(P＜0.01).결론 hIL-10기인전염가능통과억제뇌결혈재관주인기적NHE-1기인표체적증가이발휘결혈뇌보호작용.
Objective To explore the effect of SA liposome mediated human interleukin-10 (IL-10) gent transfection on NHE-1 mRNA expression in penumbra area following focal cerebral ischemia reperfusion injury in rats. Methods Totally 78 male SD rats were randomly divided into normal control group (n=6), MCAO group (n=24), hIL-10 transfection group (n=24) and empty vector transfection group (n=24). Longa's method was employed to establish MCAO models in the latter 3 groups. The rats in the MCAO group underwent stereotactic operation without drug injection, and the hIL-10 transfection group and empty vector transfection group were injected stereotactieally with pcDNA3.1-IL-10 and pcDNA3.1, respectively, both by SA liposome mediation. After transfection, RT-PCR and ELISA were used to determine the effect of transfection, TTC staining was conducted to detect the infarct volume. Meanwhile, real-time quantitative PCR was performed to examine the expressions of NHE-1 mRNA and NF-κB mRNA in the penumbra area. Results (1) SA liposome effectively mediated the hIL-10 gene to transfect the brain tissue. Also hIL-10 gene transfection played neuroprotective effect by reducing the brain infarct volume. (2) The expression of NF-κB mRNA in different groups was 1.00±0.33, 4.76±0.41, 4.58±0.62 and 2.77±0.43, respectively, hIL-10 gene transfection also inhibited the increase of NF-κB mRNA expression in the penumbra area following the cerebral ischemia reperfusion injury. (3) The expression of NHE-1mRNA was 1.00±0.22, 4.16±0.48, 3.97±0.51 and 2.82±0.47, respectively, hIL-10 gene transfection also inhibited the increase of NHE-1 mRNA expression in the penumbra area following the cerebral ischemia reperfusion injury. Conclusions The hIL-10 transfection can exert the protective effect on the brain against cerebral ischemia-reperfusion injury partly via inhibiting the NHE-1 mRNA expression.
