CAJ | 학술논문

目的观察1,6-二磷酸果糖(FDP)对病毒性心肌炎小鼠心肌还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶p22phox亚基蛋白表达的影响,探讨FDP对病毒性心肌炎的保护作用.方法 4周龄雌性BALB/c小鼠30只,体质量(12±2)g.按体质量将小鼠随机分为病毒组和治疗组,每组15只.两组小鼠同时1次性腹腔注射柯萨奇B3病毒(CVB3) 0.1 ml,治疗组在注射CVB3后的第1天,每日腹腔注射1次FDP,连续注射7d,注射剂量为300 mg/kg.在注射结束后的第4、8、21天,两组分别各处死5只小鼠,取心脏做心肌病理检查,Western免疫印迹法检测病毒性心肌炎小鼠心肌细胞内NADPH氧化酶p22phox亚基蛋白表达,图像分析系统测量p22phox亚基蛋白阳性表达区域平均吸光度(A)值,并进行定量分析.结果感染后第4天,光镜下病毒组小鼠心肌间可见少量炎症细胞浸润,心肌细胞肿胀；治疗组小鼠心肌间仅有少量炎性细胞浸润.感染后第8天,病毒组小鼠心肌出现坏死性崩解,大量炎症细胞浸润；治疗组小鼠心肌间可见稀疏的散在炎性细胞浸润.感染后第21天,病毒组小鼠心肌坏死灶中有慢性炎症细胞浸润,出现结缔组织增生;治疗组小鼠心肌可见少量慢性炎症细胞浸润.病毒组在第8天炎症浸润最严重.在病毒感染后第4、8、21天,治疗组心肌病变积分[(0.88±0.23)、(2.20±0.24)、(1.56±0.17)分]低于病毒组[(1.32±0.12)、(3.0±0.25)、(2.04±0.17)分,t值分别为3.793、5.1645、4.457,P均＜0.01].Western免疫印迹法分析结果显示,在病毒感染后第4、8、21天,治疗组NADPH氧化酶p22phox亚基蛋白表达(0.776±0.017、0.751±0.018、0.689±0.034)明显低于病毒组(1.052±0.015、0.952±0.019、0.907±0.025,t值分别为3.391、6.716、2.750,P均＜0.01或＜0.05).结论 FDP能下调NADPH氧化酶p22phox亚基蛋白表达,FDP可能通过改变NADPH酶的表达对心肌发挥保护性作用.
목적 관찰1,6-이린산과당(FDP)대병독성심기염소서심기환원형연선알선표령이핵감산린산(NADPH)양화매p22phox아기단백표체적영향,탐토FDP대병독성심기염적보호작용.방법 4주령자성BALB/c소서30지,체질량(12±2)g.안체질량장소서수궤분위병독조화치료조,매조15지.량조소서동시1차성복강주사가살기B3병독(CVB3) 0.1 ml,치료조재주사CVB3후적제1천,매일복강주사1차FDP,련속주사7d,주사제량위300 mg/kg.재주사결속후적제4、8、21천,량조분별각처사5지소서,취심장주심기병리검사,Western면역인적법검측병독성심기염소서심기세포내NADPH양화매p22phox아기단백표체,도상분석계통측량p22phox아기단백양성표체구역평균흡광도(A)치,병진행정량분석.결과 감염후제4천,광경하병독조소서심기간가견소량염증세포침윤,심기세포종창；치료조소서심기간부유소량염성세포침윤.감염후제8천,병독조소서심기출현배사성붕해,대량염증세포침윤；치료조소서심기간가견희소적산재염성세포침윤.감염후제21천,병독조소서심기배사조중유만성염증세포침윤,출현결체조직증생;치료조소서심기가견소량만성염증세포침윤.병독조재제8천염증침윤최엄중.재병독감염후제4、8、21천,치료조심기병변적분[(0.88±0.23)、(2.20±0.24)、(1.56±0.17)분]저우병독조[(1.32±0.12)、(3.0±0.25)、(2.04±0.17)분,t치분별위3.793、5.1645、4.457,P균＜0.01].Western면역인적법분석결과현시,재병독감염후제4、8、21천,치료조NADPH양화매p22phox아기단백표체(0.776±0.017、0.751±0.018、0.689±0.034)명현저우병독조(1.052±0.015、0.952±0.019、0.907±0.025,t치분별위3.391、6.716、2.750,P균＜0.01혹＜0.05).결론 FDP능하조NADPH양화매p22phox아기단백표체,FDP가능통과개변NADPH매적표체대심기발휘보호성작용.
Objective To approach the effect of fructose-1,6-diphosphate(FDP) on nicotinamide adenine dinucleotide phosphate(NADPH) oxidase p22phox subunit expression in myocardial cells of viral myocarditis mouse model and to explore the role of FDP in viral myocarditis.Method Thirty 4-week-old female BALB/c mice weighting(12 ± 2)g were randomly divided into virus group and treatment group(n =15) according to body mass.The two groups of mice were given Coxsackie B3 virus(CVB3) 0.1 ml through a single peritoneal injection.One day after the injection of CVB3,the treatment group was given FDP 300 mg/kg body weight through peritoneal injection daily,which continued for 7 days.The 4th,8th and 21th days after the injection,5 mice were sacrificed in each group,hearts were taken for myocardial pathological examination and the expression of NADPH oxidase p22phox subunit protein in myocardial cells of viral myocarditis mice was detected by Western blotting.The average absorbance(A) value of p22phox subunit proteins was measured by image analysis system and quantitative analysis was carried out.Results The 4th days after infection,a small amount of inflammatory cell infiltration and myocardial cell swelling could be seen in the virus infection group while only a small amount of inflammatory cell infiltration could be seen in the treatment group.The 8th days after infection,myocardial necrotic disintegration,and inflammatory cell infiltration could be seen in the virus infection group while sparsely scattered inflammatory cell infiltration was seen in the treatment group.The 21th days after infection,chronic inflammatory cell infiltration,connective tissue proliferation appeared in the virus infection group while a small number of chronic inflammatory cells infiltration appeared in the treatment group.The most serious inflammatory infiltrate appeared on the 8th day.The myocardial histopathology scores of the treatment group[(0.88 ± 0.23),(2.20 ± 0.24),(1.56 ± 0.17)score],the 4th,8th,21th days after infection,were all lower than that of the virus infection group [(1.32 ± 0.12),(3.00 ±0.25),(2.04 ± 0.17)score; t =3.793,5.165,4.457,all P ＜ 0.01]; the Western blotting analysis showed that,the 4th,8th,21th days after infection,the expression of NADPH oxidase subunit p22phox protein of the treatment group(0.776 ± 0.017,0.751 ± 0.018,0.689 ± 0.034) was significantly lower than that of the virus infection group (1.052 ± 0.015,0.952 ± 0.019,0.907 ± 0.025; t =3.391,6.716,2.750,P ＜ 0.01 or ＜ 0.05).Conclusions FDP can down regulate the expression of NADPH oxidase p22phox subunit protein.FDP may play a protective role through regulating the expression of NADPH oxidase.