扁塑藤素对乙型病毒性肝炎大鼠免疫功能以及纤维化蛋白表达影响

Effect of pristimerin on the immune function and expression of fibrosis protein in rats with hepatitis B virus

目的 探讨扁塑藤素对乙型病毒性肝炎(乙肝)大鼠免疫功能以及纤维化蛋白表达影响。方法 SD大鼠分成对照组、模型组(注射乙肝病毒)、实验低剂量组(注射乙肝病毒,1 mg·kg^(-1)的扁塑藤素治疗)、实验中剂量组(注射乙肝病毒,2 mg·kg^(-1)的扁塑藤素治疗)、实验高剂量组(注射乙肝病毒,4 mg·kg^(-1)的扁塑藤素治疗)、阳性对照组(注射乙肝病毒,拉米夫定治疗)。以比色法检测血清中谷丙转氨酶(GPT)、谷草转氨酶(GOT)含量,以流式细胞术检测T淋巴细胞亚群,以酶联免疫吸附(ELISA)法检测血清中干扰素-γ(IFN-γ)、白细胞介素-2(IL-2)、白细胞介素-4(IL-4)含量,用蛋白质印迹法检测转化生长因子β1(TGF-β1)、纤维连接蛋白(FN)、α-平滑肌肌动蛋白(α-SMA)和p65蛋白表达。结果 对照组、模型组、实验低剂量组、实验中剂量组、实验高剂量组、阳性对照组血清中GPT含量为(46.59±2.93),(87.12±10.84),(74.55±3.74),(61.21±6.21),(53.68±5.15),(64.59±5.43)U·L^(-1);GOT含量为(50.66±3.96),(94.61±3.78),(76.43±8.33),(63.38±5.76),(56.56±6.84),(69.07±6.08)U·L^(-1);T淋巴细胞亚群CD3^(+)比例为(21.37±2.29)%,(13.70±1.92)%,(15.56±1.22)%,(17.07±1.44)%,(20.54±1.46)%,(17.53±1.60)%;CD4^(+)/CD8^(+)比值为1.83±0.20,0.43±0.06,0.53±0.08,0.76±0.06,1.28±0.18,0.71±0.08;血清中IFN-γ含量为(53.49±5.44),(19.90±2.02),(30.79±3.50),(38.58±2.89),(45.92±5.84),(30.34±3.17)pg·mL^(-1);IL-2含量为(79.19±9.19),(39.16±4.41),(44.77±3.90),(51.32±5.46),(61.94±6.28),(54.70±4.53)pg·mL^(-1);IL-4含量为(8.79±0.83),(44.16±3.32),(36.91±2.78),(27.01±3.30),(15.98±2.09),(24.10±1.75)pg·mL^(-1),上述指标,模型组与对照组比,差异均有统计学意义(均P<0.05);实验低剂量组、实验中剂量组、实验高剂量组、阳性对照组与模型组比,差异均有统计学意义(均P<0.05);实验中、低剂量组相比,实验高、中剂量组相比,差异均有统计学意义(均P<0.05)。结论 扁塑藤素能够改善乙肝大鼠免疫功能,抑制纤维化蛋白表达,机制可能与NF-κB信号有关。

Objective To investigate the effect of pristimerin on the immune function and the expression of fibrosis protein in rats with hepatitis B virus.Methods SD rats were divided into control group,model group (injected with hepatitis B virus),experimental-L group(injection of hepatitis B virus,1 mg·kg^(-1)pristimerin),experimental-M group (injected with hepatitis B virus,2 mg·kg^(-1)pristimerin),experimental-H group (injected with hepatitis B virus,4 mg·kg^(-1)pristimerin treatment),positive control group (injected with hepatitis B virus,lamivudine treatment).Colorimetric method was used to detect serum glutamic-pyruvic transaminase (GPT) and glutamicoxaloacetic transaminase (GOT) content;flow cytometry was used to detect T lymphocyte subsets;enzyme-linked immunosorbent assay method was used to detect serum interferon-γ (IFN-γ),interleukin-2 (IL-2),interleukin-4 (IL-4) content;Western blot method was used to detect transforming growth factor β1 (TGF-β1),fibronectin(FN),α-smooth muscle actin (α-SMA) and p65 protein expression.Results The serum GPT levels in control group,model group,experimental-L group,experimental-M group,experimental-H group,and positive control group were (46.59±2.93),(87.12±10.84),(74.55±3.74),(61.21±6.21),(53.68±5.15),(64.59±5.43)U·L^(-1);GOT content were(50.66±3.96),(94.61±3.78),(76.43±8.33),(63.38±5.76),(56.56±6.84),(69.07±6.08) U·L^(-1);the T lymphocyte subset CD3^(+)ratio were (21.37±2.29)%,(13.70±1.92)%,(15.56±1.22)%,(17.07±1.44)%,(20.54±1.46)%,(17.53±1.60)%;CD4^(+)/CD8^(+)ratio were1.83±0.20,0.43±0.06,0.53±0.08,0.76±0.06,1.28±0.18,0.71±0.08;the serum IFN-γ content were(53.49±5.44),(19.90±2.02),(30.79±3.50),(38.58±2.89),(45.92±5.84),(30.34±3.17) pg·m L-1;IL-2 content were (79.19±9.19),(39.16±4.41),(44.77±3.90),(51.32±5.46),(61.94±6.28),(54.70±4.53) pg·m L-1;IL-4 content were (8.79±0.83),(44.16±3.32),(36.91±2.78),(27.01±3.30),(15.98±2.09),(24.10±1.75) pg·m L-1;above indicators,the difference between model group and control group were statistically significant (all P<0.05);the difference between experimental-L group,experimental-M group,experimental-H group,positive control group and model group were statistically significant (all P<0.05);the difference between experimental-M and experimental-L group were statistically significant (all P<0.05);the difference between experimental-H group and experimental-M group were statistically significant (all P<0.05).Conclusion Pristimerin can improve the immune function and inhibit the expression of fibrotic protein of rats with viral hepatitis B,the mechanism may be related to NF-κB signaling.