三叶青块根不同提取物抗胸腺萎缩药效研究

Anti-thymic atrophy efficacy evaluation?of of different extracts from root tubers of Tetrastigma Hemsleyanum Diels et. Gilg

关键词:  三叶青块根  提取物  胸腺萎缩

Key words:  root tubers of Tetrastigma Hemsleyanum Diels et. Gilg  extract  thymic atrophy

摘要:

目的：基于腹腔注射聚肌苷酸-聚胞苷酸（PolyIC）建立的小鼠胸腺萎缩模型，比较三叶青块根石油醚(PEE)、乙酸乙酯(EAE)、正丁醇(BE)、水提取物(WE)抗胸腺萎缩的药效作用，初步探讨其作用机制。方法：100只ICR小鼠随机分为10组：空白对照组、模型对照组、四种提取物组分低、高剂量组，每组10只，空白对照组、模型对照组均同时给予同体积的纯化水，连续灌胃10 d。给药第9天开始，除空白对照组腹腔注射生理盐水外，其余组别均腹腔注射16mg·kg-1的PolyIC溶液，连续2天。取胸腺称重并计算脏器指数，取血进行白细胞分析及IL-2、TNF-α的检测，分析胸腺组织的苏木精-伊红染色结果，通过Western blotting检测TNF-α、P-NF-κB、NF-κB的表达。结果：相较于模型对照组，提取物各组均可不同程度提升胸腺重量和胸腺指数，及皮质和髓质面积比值和髓质1mm2的胸腺小体数量，显著降低血浆中TNF-α水平，明显抑制NF-κB的磷酸化，降低TNF-α的蛋白表达，且在不同指标上的作用强度各有优劣。WE低剂量组、EAE低剂量组和PEE高剂量组外周血WBC、LYMPH#和LYMPH%升高；WE低剂量组和BE高剂量组IL-2水平显著升高，这些作用表现尤为突出。结论：三叶青块根四种提取物可不同程度改善胸腺萎缩和胸腺组织形态改变，调节造模后动物外周血白细胞失衡，可能与抑制PolyIC诱导转录因子NF-κB的磷酸化、降低促炎因子的蛋白表达有关。

Abstract:

OBJECTIVE Comparison of the anti-thymic atrophy effects of ?petroleum?ether(PEE),?ethyl?acetate(EAE),?n-butanol(BE)and?water?extract(WE)fractions from root tubers of Tetrastigma Hemsleyanum Diels et. Gilg based on mice thymic atrophy model established by intraperitoneal injection of PolyIC to preliminarily investigate the mechanisms of its effects. METHODS 100 ICR mice were randomly divided into 10 groups: a blank control group, a model control group, and low and high dose groups of the four extract fractions, with 10 mice in each group,the control group and model group were given equal volume of purified water and gavaged for 10 days.Starting on the 9th day, PolyIC solution of 16 mg·kg-1 was injected intraperitoneally for 2 consecutive days, except for the blank control group, which was injected intraperitoneally with physiological saline.Samples of thymics?were?weighed?and and?visceral?index?was?calculated, blood?samples were taken?for?analysis?of IL-2,TNF-α and WBC,thymic histopathology?was?analyzed using?hematoxylin-eosin?staining,Western?blot?was performed to detect the expression of TNF-α,P-NF-κB and NF-κB.?RESULTS Compared with the model control group,all groups of extracts could enhance the thymus weight and thymus index, as well as the cortex and medulla area ratio and the number of thymic corpuscles in 1 mm2 of medulla to different degrees，and at the same time, significantly reduce the level of TNF-α in plasma，evidently inhibited the phosphorylation of NF-κB, and reduced the protein expression of TNF-α,and each?of?them?having?distinct?advantages?and?disadvantages.Peripheral blood WBC, LYMPH# and LYMPH% were elevated in the low-dose?WE?group,low-dose?EAE?group and high dose PEE group,IL-2 levels were significantly elevated in the low-dose WE group and high dose BE group,and these effects were particularly well demonstrated. CONCLUSION Four extracts from root tubers of Tetrastigma Hemsleyanum Diels et. Gilg can?improve morphological?changes and atrophy?of the thymus?tissue?in different?degrees,regulate of peripheral blood leukocyte imbalance in post-modeling mice,it may be related to the inhibition of PolyIC-induced phosphorylation of the NF-κB and reduction of protein expression of pro-inflammatory factors.