摘要
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）0．1ml，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300mg／kg。在注射结束后的第4、8、2l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∽）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〈0．0l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。
Objective To approach the effect of fructose-i, 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.
出处
《中华地方病学杂志》
CAS
CSCD
北大核心
2013年第1期50-53,共4页
Chinese Journal of Endemiology
基金
黑龙江省自然科学基金
