阿苯达唑脂质体及槐耳浸膏对小鼠肝细粒棘球蚴感染率影响的实验观察

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目的探讨阿苯达唑脂质体(L-ABZ)及槐耳浸膏(HEF)对小鼠肝细粒棘球蚴病术后感染的抑制作用,了解增强宿主免疫功能对本病术后复发的影响。方法雌性昆明小鼠,腹腔注射人肝细粒棘球蚴囊液,3周后取外周血IgG抗体阳性鼠,分为药物治疗组(A、B、C组)和模型对照组(D组),每组40只。A组灌胃L-ABZ(75mg/kg)、B组灌胃HEF(15000mg/kg)、C组两药联用(阿75mg/kg+槐15000mg/kg)。隔天1次,共3次。麻醉各组小鼠,打开腹腔,肝脏注射经PBS浸泡20min的原头节(0.3ml/只,约含原头节6000个,下同),模拟开放式外膜内外囊完整摘除术术中外溢原头节再感染。缝合关闭腹腔,72h后药物治疗组同法继续治疗1个月,D组灌服蒸馏水0.3ml/只。E组(8只正常小鼠)为空白对照。各组小鼠均于3个月后解剖。评价药物疗效(包括肝棘球蚴复发率、组织学观察、测定脾脏指数(SI)及IgG、IgE水平、流式细胞仪检测外周血CD4+、CD8+T淋巴细胞百分率)。另取上述外周血IgG抗体阳性鼠分为F、G、H和I4组,每组约30只,分别同上述A、B、C和D组同法灌胃治疗,于肝脏分别注射经20%NaCl、75%乙醇、L-ABZ及PBS浸泡20min的原头节,观察其感染率。结果C组肝棘球蚴复发率为5.7%,低于A组(17.1%)和B组(24.2%);棘球蚴色泽发白呈结节状,生发层和角质层破坏严重。脾脏指数(A组为3.84±0.86、B组为3.95±1.01、C组为3.27±0.52)和IgE水平(A组为0.06±0.08、B组为0.07±0.08、C组为0.03±0.03)均明显低于D组(分别为5.46±0.52及0.20±0.02)(P<0.05),其中C组降低最为明显。CD8+水平,A组为16.61±3.89、B组为18.18±3.90、C组为15.38±2.63,均明显低于D组的32.90±4.71(P<0.05)。CD4+/CD8+,A组为3.21±0.70、B组为3.05±0.66、C组为3.53±0.57,均明显高于D组的1.57±0.26(P<0.05)。C组CD8+降低、CD4+/CD8+升高最为明显。F、G和H组肝棘球蚴感染率分别为0、0及23.1%,与I组(31.2%)相比较,差异有统计学意义(P<0.01)。结论HEF可明显增强小鼠免疫功能,与L-ABZ联用可抑制棘球蚴生长,降低术后复发率。L-ABZ在术中对于原头节的杀灭作用不及20%NaCl和75%乙醇。 Objective To investigate the inhibitory effect of albendazole liposomes (L-ABZ) and Huai Er extract (HEF) on the postoperative infection of mice with hepatocellular hydatid disease, and to understand the effect of enhancing host immune function on postoperative recurrence Impact. Methods Female Kunming mice were injected intraperitoneally with human hepatic Echinococcus granulosus. After 3 weeks, peripheral blood IgG antibody positive mice were divided into two groups: drug-treated group (group A, B and C) and model control group (group D) , Each group of 40. Group A was given L-ABZ (75mg / kg), Group B was given HEF (15000mg / kg), Group C was combined with two drugs (75mg / kg + 15000mg / kg). The next day, a total of 3 times. The mice in each group were anesthetized, the abdominal cavity and liver were injected with prolamins (0.3ml / body, containing about 6000 heads, the same below) immersed in PBS for 20min to simulate the spillover of the complete open endomyocardial and external capsule The original head and then infected. Suture closed the abdominal cavity, 72h after drug treatment group with the law continued treatment for 1 month, D group drank distilled water 0.3ml / only. E group (8 normal mice) as a blank control. All mice were dissected after 3 months. Evaluation of drug efficacy (including the recurrence rate of liver hydatid cyst, histological observation, determination of spleen index (SI) and IgG, IgE levels, flow cytometry detection of peripheral blood CD4 +, CD8 + T lymphocyte percentage). Another group of positive peripheral blood IgG antibodies were divided into F, G, H and I4 groups of about 30, respectively, with the above groups A, B, C and D with the same gavage treatment in the liver were injected with 20% NaCl, 75% ethanol, L-ABZ and PBS soaking for 20 minutes to observe the infection rate. Results The recurrence rate of Echinococcus granulosus in group C was 5.7%, which was lower than that in group A (17.1%) and group B (24.2%). Echinococcus granulosus developed nodular color with serious degeneration of germinal layer and stratum corneum. Spleen index (3.84 ± 0.86 in group A, 3.95 ± 1.01 in group B and 3.27 ± 0.52 in group C) and IgE levels (0.06 ± 0.08 in group A, 0.07 ± 0.08 in group B and 0.03 ± 0.03 in group C) Significantly lower than those in group D (5.46 ± 0.52 and 0.20 ± 0.02, respectively) (P <0.05), with the most obvious decrease in group C. CD8 + level in group A was 16.61 ± 3.89, in group B was 18.18 ± 3.90, in group C was 15.38 ± 2.63, which were significantly lower than those in group D (32.90 ± 4.71, P <0.05). CD4 + / CD8 + was 3.21 ± 0.70 in group A, 3.05 ± 0.66 in group B and 3.53 ± 0.57 in group C, respectively, which were significantly higher than those in group D (1.57 ± 0.26, P <0.05). C group CD8 + decreased, the most obvious increase of CD4 + / CD8 +. The infection rates of Echinococcus granulosus in group F, G and H were 0, 0 and 23.1%, respectively, which were significantly different from those in group I (31.2%) (P <0.01). Conclusion HEF can significantly enhance the immune function in mice, and combined with L-ABZ can inhibit the growth of hydatid cyst and reduce the recurrence rate. L-ABZ killed less than 20% NaCl and 75% ethanol during the operation.
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