赤芍煎剂对急性胰腺炎大鼠细胞因子表达的影响

目的 观察赤芍煎剂对急性胰腺炎(AP)大鼠的治疗效果及对NF-κB,TNF-α,IL-6表达水平的影响.方法SD大鼠40只随机分为5组:正常对照组(A组)、AP模型组24 h点(B组)、AP模型组72 h点(C组)、赤芍煎剂治疗组24 h点(D组)和赤芍煎剂治疗组72 h点(E组),每组8只.采用L-精氨酸腹腔内注射的方法诱发急性胰腺炎模型,造模后立即灌胃生理盐水(B,C组)、赤芍煎剂(D,E组),于造模后24 h,72 h时间点处死各组大鼠.采血检测血清淀粉酶;取胰腺组织分3份:HE染色行光镜检查、Western Blot法检测NF-κB p65蛋白表达、实时荧光定量PCR技术进行TNF-α、IL-6的mRNA定量分析.结果 造模24 h后与A组比较,B,D组大鼠血清淀粉酶均升高(P<0.05).D组与B组比较淀粉酶水平明显降低(P<0.05).造模后72 h,E,C组淀粉酶水平均下降接近正常组水平(P>0.05).D,E组较同时间点B,C组的病理改变和病理学评分均明显减轻(P<0.05).Western blotting结果显示,D,E组较同时间点B,C组NF-κB p65蛋白的表达明显减少D组与B组相比,(0.07±0.006)vs.(0.71±0.029),P<0.05;E组与C组相比,(0.18±0.014)vs.(0.65±0.026),P<0.05].实时荧光定量PCR结果显示:模型组TNF-α,IL-6 mRNA的表达明显高于赤芍煎剂治疗组(P<0.05).结论赤芍煎剂对AP大鼠治疗作用的机制可能与其抑制NF-κB活化、在整体水平上抑制细胞因子基因表达、使机体免受过量细胞因子的损伤等因素有关.

Effects of Radix Paeoniae Rubra decoction on the expression of cvtokines in rats with acute pancreatitis

Objective To observe the therapeutic effects of Radix Paeoniae Rubra decoction (RPRD) on a-cute pancreatitis (AP) in rats, and to study effects of RPRD on the expression of NF-κB, TNF-α and IL-6. Method Forty Sprague-Dawley (SD) rats were, randomly divided into five groups: normal control group (group A), AP model groups (group B, 24 h; group C,72 h) and RPRD treatment groups (group D, 24 h;group E,72 h).There were 8 rats in each group. The AP models were induced by intraperitoneally injecting with L-arginine. After the model was estabhshed, rats of group B and group C were treated with inn'a-gastric instillation of NS, and rats in group D and group E with RPRD. At 24 and 72 hours after induction of AP, rats in each group were sacri-riced and blood samples were taken for the measurement of serum amylase. The expression of NF-κB 1065 protein and TNF-α and IL-6 mRNA in the pancreas was detected by Western blotting and real-time quantitative PCR, re-spectively, and the pathological status of the pancreas was analyzed. Results The serum amylase level in group B and group D was obviously increased at 24 hours, and were significantly higher than that in group A (P < 0.05) ; the level in group D was lower thaa that in group B (P < 0.05). the level reduced at 72 hours, and there was no difference between group E and group C (P > 0.05). The pathological severity and the pathological score of pan-creatic tissue in group D and group E at 24 hours and 72 hours were more hghter than those in group B and hours C (P < 0. 05). The expression levels of NF-κB p65 protein (Western blotting) in the pancreas of group D and group E were significantly lighter than those in group B and group C at both time points (group D compared with group B, (0.07 ± 0.006) vs. (0.71 ± 0.029), P < 0.05;group D compared with B, (0.07 ± 0.006) vs. (0.71 ±0.029), P <0.05; group E compared with C, 0.18±0.014 vs. 0.65±0.026, P <0.05). The results of real-time quantitative PCR shown that the expression levels of TNF-α and IL-6 mRNA in model treatment group were significantly stronger than that in RPRD group at the two time points (P < 0.05). Conclusions RPRD is effective for the treatment of AP rats to inhibit the activity of NF-κB. Inhibition of NF-κB offers a promising strategy to treat AP by down-regulating eytokine expression.