中华心血管病杂志
中華心血管病雜誌
중화심혈관병잡지
Chinese Journal of Cardiology
2013年
9期
778-784
,共7页
王仲华%戴刚%周荣靓%匡泽民
王仲華%戴剛%週榮靚%劻澤民
왕중화%대강%주영정%광택민
心肌梗死%杀伤细胞,天然%白细胞介素2%新生血管化,生理性
心肌梗死%殺傷細胞,天然%白細胞介素2%新生血管化,生理性
심기경사%살상세포,천연%백세포개소2%신생혈관화,생이성
Myocardial infarction%Killer cells,natural%Interleukin-2%Neovascularization,physiologic
目的 验证心肌内注射重组人白细胞介素-2(rhIL-2)活化后的自然杀伤细胞(NK)是否能促进心肌梗死后血管新生和保护心肌功能.方法 体外提取NK细胞并使用rhIL-2活化,以未活化NK细胞为对照,测定rhIL-2-NK的杀伤能力.rhIL-2-NK与心肌微血管内皮细胞共培养,以5％多聚甲醛固定后的rhIL-2-NK为阴性对照组,NK细胞单独培养作为空白对照组,观察rhIL-2-NK对心肌微血管内皮细胞增殖的影响.大鼠心肌梗死后1h,随机分为3组,分别为rhIL-2-NK组、NK组、空白对照组,分别在其梗死区域注射NK、rhIL-2-NK和PBS,选取0、1、3、5、7和20 d6个时间点,通过定量PCR分析各组巨噬细胞趋化性和激活性因子(MCP-1)、肿瘤坏死因子-α(TNF-α)、白细胞介素-2(IL-2)mRNA的表达水平.治疗结束后,免疫组织化学检测各组大鼠血小板内皮细胞黏附分子(CD31)和血管内皮生长因子(VEGF)的表达水平,并通过超声心动图检查、血液动力学测定、组织形态学分析来检测各组大鼠的心功能.结果 体外成功提取具有杀伤力的NK细胞,rhIL-2-NK体外促进心肌微血管内皮细胞的增殖,rhIL-2-NK组增殖数量比空白对照高40％ (P ＜0.01).在大鼠心肌梗死治疗中,rhIL-2-NK组大鼠心肌MCP-1、TNF-α、IL-2、CD31和VEGF mRNA的表达水平均明显高于空白对照组(P均＜0.01).超声心动图及组织形态学显示rhIL-2-NK组大鼠心功能得到改善,微血管密度平均为17.35±1.82,明显高于空白对照组的4.76±0.92 (P＜0.01);左心室射血分数平均为(77.56±15.67)％,与假手术组[(85.35±13.34)％]接近,并高于空白对照组[(41.47±12.21)％,P＜0.05].结论 在大鼠梗死心肌内注射rhIL-2-NK可促进血管新生,并最终改善心功能.
目的 驗證心肌內註射重組人白細胞介素-2(rhIL-2)活化後的自然殺傷細胞(NK)是否能促進心肌梗死後血管新生和保護心肌功能.方法 體外提取NK細胞併使用rhIL-2活化,以未活化NK細胞為對照,測定rhIL-2-NK的殺傷能力.rhIL-2-NK與心肌微血管內皮細胞共培養,以5％多聚甲醛固定後的rhIL-2-NK為陰性對照組,NK細胞單獨培養作為空白對照組,觀察rhIL-2-NK對心肌微血管內皮細胞增殖的影響.大鼠心肌梗死後1h,隨機分為3組,分彆為rhIL-2-NK組、NK組、空白對照組,分彆在其梗死區域註射NK、rhIL-2-NK和PBS,選取0、1、3、5、7和20 d6箇時間點,通過定量PCR分析各組巨噬細胞趨化性和激活性因子(MCP-1)、腫瘤壞死因子-α(TNF-α)、白細胞介素-2(IL-2)mRNA的錶達水平.治療結束後,免疫組織化學檢測各組大鼠血小闆內皮細胞黏附分子(CD31)和血管內皮生長因子(VEGF)的錶達水平,併通過超聲心動圖檢查、血液動力學測定、組織形態學分析來檢測各組大鼠的心功能.結果 體外成功提取具有殺傷力的NK細胞,rhIL-2-NK體外促進心肌微血管內皮細胞的增殖,rhIL-2-NK組增殖數量比空白對照高40％ (P ＜0.01).在大鼠心肌梗死治療中,rhIL-2-NK組大鼠心肌MCP-1、TNF-α、IL-2、CD31和VEGF mRNA的錶達水平均明顯高于空白對照組(P均＜0.01).超聲心動圖及組織形態學顯示rhIL-2-NK組大鼠心功能得到改善,微血管密度平均為17.35±1.82,明顯高于空白對照組的4.76±0.92 (P＜0.01);左心室射血分數平均為(77.56±15.67)％,與假手術組[(85.35±13.34)％]接近,併高于空白對照組[(41.47±12.21)％,P＜0.05].結論 在大鼠梗死心肌內註射rhIL-2-NK可促進血管新生,併最終改善心功能.
목적 험증심기내주사중조인백세포개소-2(rhIL-2)활화후적자연살상세포(NK)시부능촉진심기경사후혈관신생화보호심기공능.방법 체외제취NK세포병사용rhIL-2활화,이미활화NK세포위대조,측정rhIL-2-NK적살상능력.rhIL-2-NK여심기미혈관내피세포공배양,이5％다취갑철고정후적rhIL-2-NK위음성대조조,NK세포단독배양작위공백대조조,관찰rhIL-2-NK대심기미혈관내피세포증식적영향.대서심기경사후1h,수궤분위3조,분별위rhIL-2-NK조、NK조、공백대조조,분별재기경사구역주사NK、rhIL-2-NK화PBS,선취0、1、3、5、7화20 d6개시간점,통과정량PCR분석각조거서세포추화성화격활성인자(MCP-1)、종류배사인자-α(TNF-α)、백세포개소-2(IL-2)mRNA적표체수평.치료결속후,면역조직화학검측각조대서혈소판내피세포점부분자(CD31)화혈관내피생장인자(VEGF)적표체수평,병통과초성심동도검사、혈액동역학측정、조직형태학분석래검측각조대서적심공능.결과 체외성공제취구유살상력적NK세포,rhIL-2-NK체외촉진심기미혈관내피세포적증식,rhIL-2-NK조증식수량비공백대조고40％ (P ＜0.01).재대서심기경사치료중,rhIL-2-NK조대서심기MCP-1、TNF-α、IL-2、CD31화VEGF mRNA적표체수평균명현고우공백대조조(P균＜0.01).초성심동도급조직형태학현시rhIL-2-NK조대서심공능득도개선,미혈관밀도평균위17.35±1.82,명현고우공백대조조적4.76±0.92 (P＜0.01);좌심실사혈분수평균위(77.56±15.67)％,여가수술조[(85.35±13.34)％]접근,병고우공백대조조[(41.47±12.21)％,P＜0.05].결론 재대서경사심기내주사rhIL-2-NK가촉진혈관신생,병최종개선심공능.
Objective To investigate the effect of recombinant human interleukin-2 (rhIL-2) activated natural killer cells (rhIL-2-NK) on angiogenesis and cardiac function of rats with myocardial infarction (MI).Methods Natural killer cells (NKs) were isolated and activated by rhIL-2 in vitro.Untreated NKs were used as the control,the killing capacity of rhIL-2-NK were evaluated with cytotoxicity assay.Cardiac microvascular endothelial cells (CMECs) were cocultured with rhIL-2-NK.One hour after MI,rats were randomly divided into rhIL-2-NK group,NK group and blank control group and NK,rhIL-2-NK and PBS were injected directly in the infracted myocardium.At the 0,1st,3rd,5th,7th and 20th day after MI,the mRNA expression of monocyte chemotactic protein-1 (MCP-1),Tumor necrosis factor-α (TNF-α) and interleukin-2 (IL-2) were was detected by q-PCR essay.At the end of the therapy,the platelet endothelial cell adhesion molecule-1 (CD31) and vascular endothelial growth factor (VEGF) were evaluated through immunohistochemical assay,and the cardiac function observed with echocardiography,homodynamic measurements.Results The NKs were isolated successfully and the CMEC were proliferated remarkably by coculturing with rhIL-2-NK (P ＜0.01).The mRNA expression of MCP-1,TNF-α,CD31 and rhIL-2,VEGF were significantly upregulated in rhIL-2-NK group than in the PBS control group (P ＜0.01).Four weeks after operation,LVEF was significantly higher in rhIL-2-NK group than in the PBS control group [(77.56 ± 15.67) ％ vs.(41.47 ± 12.21) ％,P ＜ 0.05)] and histomorphology assay revealed that the density of microvascular endothelial (MVD) of rhIL-2-NK group was significantly higher than that of PBS control group (17.35 ± 1.82 vs.4.76 ± 0.92,P ＜ 0.01).Conclusions Myocardial injection of rhIL-2-NK could promote angiogenesis and improve cardiac function in MI rats.
