CAJ | 학술논문

目的：糖尿病基础上发生脑缺血再灌注损伤后，缓激肽B1受体（bradyldnin B1 receptor， B1R）和缓激肽B2受体（bradykinin B2 receptor， B2R）表达及作用的研究尚少。文中旨在观察比较非糖尿病和糖尿病大鼠脑缺血再灌注后B1R和B2R的表达差异及作用变化。方法构建非糖尿病大鼠和2型糖尿病大鼠局灶性脑缺血再灌注（ ischemia／reperfusion， I／R）模型，各41只，测试大鼠体重和生化指标。按照随机数字表法分别将8只非糖尿病大鼠及8只2型糖尿病大鼠分为2组：非糖尿病对照组和非糖尿病脑I／R 24 h组，糖尿病对照组和糖尿病脑I／R 24 h组，每组4只。实时荧光定量PCR检测再灌注24 h后B1R和B2R的表达。另将33只非糖尿病大鼠及33只2型糖尿病大鼠以随机数字表法各分为4组：假手术组（ n＝6）、等渗盐水组（n＝9）、B1R拮抗剂组（n＝9）和B2R拮抗剂组（n＝9），I／R 24 h后，采用NSS评估神经功能缺损程度，TTC染色观察脑梗死体积，TUNEL染色测定细胞凋亡，Fluoro-Jade C染色检测神经元变性。结果41只糖尿病大鼠造模后3、7、14 d空腹血糖[（23．45±5．01）、（23．71±4．87）、（22．72±4．11） mmol／L]较非糖尿病大鼠[（5．77±0．75）、（6．05±0．69）、（7．15±1．09）mmol／L]明显增加（P＜0．05）；14 d时血胆固醇[（4．59±3．43）mmol／L]、胰岛素[（67．26±12．02）pmol／L]较非糖尿病大鼠[（1．58±0．37）mmol／L、（25．34±4．88）pmol／L]明显升高（P＜0．05），而三酰甘油水平差异无统计学意义（P＞0．05）。糖尿病大鼠脑I／R 24 h后脑组织中B1R mRNA水平较非糖尿病大鼠升高更加明显（P＜0．01），而B2R mRNA水平则相对下降（ P＜0．05）。非糖尿病B2 R拮抗剂组NSS评分、梗死体积百分比、损伤和凋亡细胞数目较等渗盐水组明显减少（ P＜0．05）；糖尿病B1R拮抗剂组NSS评分、梗死体积百分比、损伤和凋亡细胞数目较等渗盐水组明显减少（P＜0．05）。结论非糖尿病大鼠脑I／R损伤诱导了B2R mRNA的显著上调，抑制B2R有效地减少脑梗死体积及减轻组织细胞损伤；糖尿病大鼠的脑I／R损伤诱导了B1R mRNA表达上调，B1R拮抗剂取代B2R拮抗剂发挥神经保护作用。目的：糖尿病基礎上髮生腦缺血再灌註損傷後，緩激肽B1受體（bradyldnin B1 receptor， B1R）和緩激肽B2受體（bradykinin B2 receptor， B2R）錶達及作用的研究尚少。文中旨在觀察比較非糖尿病和糖尿病大鼠腦缺血再灌註後B1R和B2R的錶達差異及作用變化。方法構建非糖尿病大鼠和2型糖尿病大鼠跼竈性腦缺血再灌註（ ischemia／reperfusion， I／R）模型，各41隻，測試大鼠體重和生化指標。按照隨機數字錶法分彆將8隻非糖尿病大鼠及8隻2型糖尿病大鼠分為2組：非糖尿病對照組和非糖尿病腦I／R 24 h組，糖尿病對照組和糖尿病腦I／R 24 h組，每組4隻。實時熒光定量PCR檢測再灌註24 h後B1R和B2R的錶達。另將33隻非糖尿病大鼠及33隻2型糖尿病大鼠以隨機數字錶法各分為4組：假手術組（ n＝6）、等滲鹽水組（n＝9）、B1R拮抗劑組（n＝9）和B2R拮抗劑組（n＝9），I／R 24 h後，採用NSS評估神經功能缺損程度，TTC染色觀察腦梗死體積，TUNEL染色測定細胞凋亡，Fluoro-Jade C染色檢測神經元變性。結果41隻糖尿病大鼠造模後3、7、14 d空腹血糖[（23．45±5．01）、（23．71±4．87）、（22．72±4．11） mmol／L]較非糖尿病大鼠[（5．77±0．75）、（6．05±0．69）、（7．15±1．09）mmol／L]明顯增加（P＜0．05）；14 d時血膽固醇[（4．59±3．43）mmol／L]、胰島素[（67．26±12．02）pmol／L]較非糖尿病大鼠[（1．58±0．37）mmol／L、（25．34±4．88）pmol／L]明顯升高（P＜0．05），而三酰甘油水平差異無統計學意義（P＞0．05）。糖尿病大鼠腦I／R 24 h後腦組織中B1R mRNA水平較非糖尿病大鼠升高更加明顯（P＜0．01），而B2R mRNA水平則相對下降（ P＜0．05）。非糖尿病B2 R拮抗劑組NSS評分、梗死體積百分比、損傷和凋亡細胞數目較等滲鹽水組明顯減少（ P＜0．05）；糖尿病B1R拮抗劑組NSS評分、梗死體積百分比、損傷和凋亡細胞數目較等滲鹽水組明顯減少（P＜0．05）。結論非糖尿病大鼠腦I／R損傷誘導瞭B2R mRNA的顯著上調，抑製B2R有效地減少腦梗死體積及減輕組織細胞損傷；糖尿病大鼠的腦I／R損傷誘導瞭B1R mRNA錶達上調，B1R拮抗劑取代B2R拮抗劑髮揮神經保護作用。
목적：당뇨병기출상발생뇌결혈재관주손상후，완격태B1수체（bradyldnin B1 receptor， B1R）화완격태B2수체（bradykinin B2 receptor， B2R）표체급작용적연구상소。문중지재관찰비교비당뇨병화당뇨병대서뇌결혈재관주후B1R화B2R적표체차이급작용변화。방법구건비당뇨병대서화2형당뇨병대서국조성뇌결혈재관주（ ischemia／reperfusion， I／R）모형，각41지，측시대서체중화생화지표。안조수궤수자표법분별장8지비당뇨병대서급8지2형당뇨병대서분위2조：비당뇨병대조조화비당뇨병뇌I／R 24 h조，당뇨병대조조화당뇨병뇌I／R 24 h조，매조4지。실시형광정량PCR검측재관주24 h후B1R화B2R적표체。령장33지비당뇨병대서급33지2형당뇨병대서이수궤수자표법각분위4조：가수술조（ n＝6）、등삼염수조（n＝9）、B1R길항제조（n＝9）화B2R길항제조（n＝9），I／R 24 h후，채용NSS평고신경공능결손정도，TTC염색관찰뇌경사체적，TUNEL염색측정세포조망，Fluoro-Jade C염색검측신경원변성。결과41지당뇨병대서조모후3、7、14 d공복혈당[（23．45±5．01）、（23．71±4．87）、（22．72±4．11） mmol／L]교비당뇨병대서[（5．77±0．75）、（6．05±0．69）、（7．15±1．09）mmol／L]명현증가（P＜0．05）；14 d시혈담고순[（4．59±3．43）mmol／L]、이도소[（67．26±12．02）pmol／L]교비당뇨병대서[（1．58±0．37）mmol／L、（25．34±4．88）pmol／L]명현승고（P＜0．05），이삼선감유수평차이무통계학의의（P＞0．05）。당뇨병대서뇌I／R 24 h후뇌조직중B1R mRNA수평교비당뇨병대서승고경가명현（P＜0．01），이B2R mRNA수평칙상대하강（ P＜0．05）。비당뇨병B2 R길항제조NSS평분、경사체적백분비、손상화조망세포수목교등삼염수조명현감소（ P＜0．05）；당뇨병B1R길항제조NSS평분、경사체적백분비、손상화조망세포수목교등삼염수조명현감소（P＜0．05）。결론비당뇨병대서뇌I／R손상유도료B2R mRNA적현저상조，억제B2R유효지감소뇌경사체적급감경조직세포손상；당뇨병대서적뇌I／R손상유도료B1R mRNA표체상조，B1R길항제취대B2R길항제발휘신경보호작용。
Objective There is little research focusing on the expression and function of bradykinin 1 receptor ( B1R ) and bradykinin 2 receptor ( B2R) after cerebral ischemia/reperfusion on the basis of diabetes .The aim of this study was to compare the ex-pression difference and function change of B 1R and B2R in non-dia-betic and diabetic rats . Methods The cerebral ischemia/reperfu-sion model was established on 41 non-diabetic and type 2 diabetic rats, the weight and the biochemical index were measured on these two types of rats .8 non-diabetic rats and 8 diabetic rats were respec- <br> tively assigned to two groups according to random number tables:control group and I/R 24 h group, 4 in each group.Real-time PCR was performed to observe the expressions of two receptors at 24 h after reperfusion .Then, 33 non-diabetic rats and 33 diabetic rats were randomly divided into 4 groups respectively, including sham group (n=6), saline group (n=9), B1R antagonist group (n=9) and B2R antagonist group (n=9).At 24 hours after cerebral I/R, neurological deficiency was evaluated by neurological severity scores ( NSS);infarct volume was observed by TTC staining;cell apoptosis was determined by TUNEL staining;neuron degeneration was de-tected by Fluoro-Jade C staining. Results Glucoses of diabetics at 3, 7, 14 d after model establishment [(23.45 ±5.01), (23.71 ±4.87), (22.72 ±4.11) mmol/L] were obviously elevated compared with non-diabetics [(5.77 ±0.75), (6.05 ±0.69), (7.15 ±1.09) mmol/L];blood cholesterin [(4.59 ±3.43) mmol/L] and insulin [(67.26 ±12.02) pmol/L] at 14 d after model establishment were evidently incresaed in comparison to those in non-diabetics [(1.58 ±0.37) mmol/L, (25.34 ±4.88) pmol/L] (P<0.05), while no significant difference was found in the blood triglyceride of diabetics between them (P>0.05).Compared with non-diabetics, diabetics suffered from more apparent up-regulation of B1R mRNA (P<0.01) but relatively less B2R mRNA (P<0.05) at 24 h after I/R.NSS score, infarction volume, damaged and apoptotic cells in B2R antagonis-treated non-diabetic rats at 24 h after I/R conspicuously decreased compared with saline-treated non-daibetic rats.Those indicators in B1R antagonis-treated diabeics were strikingly lessened compared with saline-treated daibetics . Conclusion I/R induced distinct up-regulation of B2R mRNA in non-diabetics and inhibiton of B 2R effectively ameliorated the infarct volume and cell injury after I/R in non-diabetics; I/R induced more notable up-regulation of B1R mRNA in diabetics and B1R antagonist exerted neuroprotective effects instead of B 2R antagonist af-ter I/R in diabetics.