鮑曼不動(dòng)桿菌膿毒癥大鼠體內(nèi)膜聯(lián)蛋白A1表達(dá)情況及血必凈對其影響
發(fā)布時(shí)間：2018-08-14 14:23
【摘要】：目的：檢測鮑曼不動(dòng)桿菌膿毒癥大鼠體內(nèi)膜聯(lián)蛋白A1(Annexin A1)在不同膿毒癥時(shí)間點(diǎn)的表達(dá)情況，并以血必凈進(jìn)行治療，通過檢測相關(guān)炎癥因子腫瘤壞死因子（TNF-α）及白介素-8（IL-8），探討AnnexinA1與鮑曼不動(dòng)桿菌膿毒癥之間的關(guān)系、血必凈對AnnexinA1表達(dá)情況的影響及血必凈對鮑曼不動(dòng)桿菌膿毒癥的治療效果。 方法：將54只健康wistar大鼠隨機(jī)分為正常對照組、膿毒癥組和血必凈治療組，正常對照組腹腔注射生理鹽水，鮑曼不動(dòng)桿菌膿毒癥組腹腔注射鮑曼不動(dòng)桿菌（ATCC19606），血必凈治療組腹腔注射鮑曼不動(dòng)桿菌（ATCC19606）同時(shí)通過尾靜脈注射血必凈注射液。每組又按處死時(shí)間分為術(shù)后6h、12h、24h三個(gè)亞組,每亞組6只。處死大鼠后，取大鼠動(dòng)脈血中性粒細(xì)胞，通過免疫印跡Westernblot檢測中性粒細(xì)胞AnnexinA1的表達(dá)，取大鼠肺臟通過免疫組化Elivision法檢測肺組織中AnnexinA1的表達(dá)，分離大鼠血清通過ELISA方法檢測血清中IL-8、TNF-α的水平。 結(jié)果：1.成功地建立正常對照組、膿毒癥組和血必凈治療組的大鼠模型。 2.大鼠中性粒細(xì)胞中Annexin A1蛋白的水平在本實(shí)驗(yàn)設(shè)計(jì)的三個(gè)觀察時(shí)間點(diǎn)（6h、12h、24h）內(nèi)：膿毒癥組較正常組蛋白含量均明顯升高，差別有統(tǒng)計(jì)學(xué)意義（p0.05）；血必凈治療組較膿毒癥組蛋白含量明顯升高，差別有統(tǒng)計(jì)學(xué)意義（p0.05）。膿毒癥6h組較正常組升高，差別有統(tǒng)計(jì)學(xué)意義（p0.05）；膿毒癥24h組和膿毒癥12h組較膿毒癥6組升高，差別有統(tǒng)計(jì)學(xué)意義（p0.05）；膿毒癥12h組和膿毒癥24h組之間差別無統(tǒng)計(jì)學(xué)意義（p0.05）。 3.血清中IL-8的水平在本實(shí)驗(yàn)設(shè)計(jì)的三個(gè)觀察時(shí)間點(diǎn)（6h、12h、24h）內(nèi)：膿毒癥組IL-8的水平較正常組IL-8的水平明顯升高，差別有統(tǒng)計(jì)學(xué)意義（p0.05），血必凈組IL-8的水平較膿毒癥組IL-8的水平明顯下降，差別有統(tǒng)計(jì)學(xué)意義（p0.05）。IL-8的水平在膿毒癥12h組和膿毒癥24h組均較膿毒癥6組升高，差別有統(tǒng)計(jì)學(xué)意義（p0.05）；膿毒癥12h組和膿毒癥24h組之間差別無統(tǒng)計(jì)學(xué)意義（p0.05）。 4.血清中TNF-α的水平在本實(shí)驗(yàn)設(shè)計(jì)的三個(gè)觀察時(shí)間點(diǎn)（6h、12h、24h）內(nèi)：膿毒癥組TNF-α的水平較正常組TNF-α的水平明顯升高，差別有統(tǒng)計(jì)學(xué)意義（p0.05），血必凈組TNF-α的水平較膿毒癥組TNF-α的水平明顯下降，差別有統(tǒng)計(jì)學(xué)意義（p0.05）。TNF-α的水平在膿毒癥12h組和膿毒癥24h組均較膿毒癥6組升高，差別有統(tǒng)計(jì)學(xué)意義（p0.05）；膿毒癥12h組和膿毒癥24h組之間TNF-α的水平差別無統(tǒng)計(jì)學(xué)意義（p0.05）。 5.在正常肺組織中，AnnexinA1表達(dá)的陽性率較低。在膿毒癥組中AnnexinA1表達(dá)的陽性率較正常肺組織升高，與正常組比較，有統(tǒng)計(jì)學(xué)差異（p0.05）；血必凈治療組（6h、12h、24h）AnnexinA1部分呈現(xiàn)強(qiáng)陽性表達(dá)，與膿毒癥組比較有統(tǒng)計(jì)學(xué)差異（p0.05）。 結(jié)論：1、AnnexinA1在膿毒癥大鼠中性粒細(xì)胞及肺臟內(nèi)，，隨著病情發(fā)展逐漸升高，并在12h至24h之間到達(dá)平衡；血必凈注射液干預(yù)后膿毒癥大鼠后，可以提高大鼠體內(nèi)AnnexinA1的水平。 2、TNF-α及IL-8在膿毒癥大鼠血清中的水平，隨著病情發(fā)展逐漸升高；血必凈注射液干預(yù)后膿毒癥大鼠后，可以降低TNF-α及IL-8在血清中的水平。 3、血必凈注射液可以有效的改善鮑曼不動(dòng)桿菌膿毒癥大鼠的病情，顯著地抑制炎癥介質(zhì)IL-8、TNF-α的過度釋放；同時(shí)可以提高體內(nèi)AnnexinA1的含量，但AnnexinA1在機(jī)體內(nèi)影響IL-8、TNF-α水平的作用機(jī)制還需進(jìn)一步證實(shí)。
[Abstract]:Objective: To detect the expression of annexin A1 (Annexin A1) in rats with Acinetobacter baumannii sepsis at different time points and treat with Xuebijing. To explore the relationship between Annexin A1 and Acinetobacter baumannii sepsis, Xuebijing was used to treat the sepsis. The effect of Xuebijing on the expression of AnnexinA1 and sepsis caused by Acinetobacter Bauman.
Methods: Fifty-four healthy Wistar rats were randomly divided into normal control group, sepsis group and Xuebijing treatment group. Normal control group was given intraperitoneal injection of normal saline, Acinetobacter baumannii (ATCC19606) was given intraperitoneal injection in Acinetobacter baumannii sepsis group, and Acinetobacter baumannii (ATCC19606) was given intraperitoneal injection in Xuebijing treatment group. Each group was divided into three subgroups 6 hours, 12 hours and 24 hours after the operation. After the rats were sacrificed, the expression of Annexin A1 was detected by Western blot, and the expression of Annexin A1 was detected by immunohistochemical Elivision. Serum levels of IL-8 and TNF- alpha were detected by ELISA in rat serum.
Results: 1. the rats in the control group, sepsis group and Xuebijing group were successfully established.
2. The level of Annexin A1 protein in rat neutrophils was significantly higher in sepsis group than that in normal group (p0.05), and significantly higher in Xuebijing treatment group than that in sepsis group (p0.05). Compared with the normal group, the difference was statistically significant (p0.05); sepsis 24 h group and sepsis 12 h group were higher than sepsis 6 group, the difference was statistically significant (p0.05); sepsis 12 h group and sepsis 24 h group had no significant difference (p0.05).
3. The level of IL-8 in serum was significantly higher in sepsis group than in normal group (p0.05). The level of IL-8 in Xuebijing group was significantly lower than that in sepsis group (p0.05). The level of IL-8 in sepsis group was significantly lower than that in sepsis group (p0.05). There was no significant difference between sepsis 12 h group and sepsis 24 h group (p0.05).
4. The level of TNF-alpha in serum was significantly higher in sepsis group than in normal group (p0.05). The level of TNF-alpha in Xuebijing group was significantly lower than that in sepsis group (p0.05). The difference was statistically significant (p0.05). The levels of TNF-alpha in 12-hour sepsis group and 24-hour sepsis group were significantly higher than those in 6-hour sepsis group (p0.05). There was no significant difference in the levels of TNF-alpha between 12-hour sepsis group and 24-hour sepsis group (p0.05).
5. In normal lung tissues, the positive rate of Annexin A1 expression was lower. In sepsis group, the positive rate of Annexin A1 expression was higher than that in normal lung tissues, and there was statistical difference (p0.05). In Xuebijing treatment group (6h, 12h, 24h), the positive rate of Annexin A1 expression was stronger than that in sepsis group (p0.05).
Conclusion: 1. The level of Annexin A1 in the neutrophils and lungs of septic rats increased gradually with the development of the disease and reached a balance between 12 and 24 hours.
2. The levels of TNF-alpha and IL-8 in the serum of septic rats increased gradually with the development of the disease; Xuebijing injection could reduce the levels of TNF-alpha and IL-8 in the serum of septic rats after intervention.
3. Xuebijing injection can effectively improve the condition of rats with Acinetobacter baumannii sepsis, significantly inhibit the excessive release of inflammatory mediators IL-8 and TNF-alpha, and increase the content of Annexin A1 in vivo, but the mechanism of Annexin A1 affecting the level of IL-8 and TNF-alpha in vivo needs further confirmation.
【學(xué)位授予單位】：蚌埠醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R459.7
本文編號：2183138
[Abstract]:Objective: To detect the expression of annexin A1 (Annexin A1) in rats with Acinetobacter baumannii sepsis at different time points and treat with Xuebijing. To explore the relationship between Annexin A1 and Acinetobacter baumannii sepsis, Xuebijing was used to treat the sepsis. The effect of Xuebijing on the expression of AnnexinA1 and sepsis caused by Acinetobacter Bauman.
Methods: Fifty-four healthy Wistar rats were randomly divided into normal control group, sepsis group and Xuebijing treatment group. Normal control group was given intraperitoneal injection of normal saline, Acinetobacter baumannii (ATCC19606) was given intraperitoneal injection in Acinetobacter baumannii sepsis group, and Acinetobacter baumannii (ATCC19606) was given intraperitoneal injection in Xuebijing treatment group. Each group was divided into three subgroups 6 hours, 12 hours and 24 hours after the operation. After the rats were sacrificed, the expression of Annexin A1 was detected by Western blot, and the expression of Annexin A1 was detected by immunohistochemical Elivision. Serum levels of IL-8 and TNF- alpha were detected by ELISA in rat serum.
Results: 1. the rats in the control group, sepsis group and Xuebijing group were successfully established.
2. The level of Annexin A1 protein in rat neutrophils was significantly higher in sepsis group than that in normal group (p0.05), and significantly higher in Xuebijing treatment group than that in sepsis group (p0.05). Compared with the normal group, the difference was statistically significant (p0.05); sepsis 24 h group and sepsis 12 h group were higher than sepsis 6 group, the difference was statistically significant (p0.05); sepsis 12 h group and sepsis 24 h group had no significant difference (p0.05).
3. The level of IL-8 in serum was significantly higher in sepsis group than in normal group (p0.05). The level of IL-8 in Xuebijing group was significantly lower than that in sepsis group (p0.05). The level of IL-8 in sepsis group was significantly lower than that in sepsis group (p0.05). There was no significant difference between sepsis 12 h group and sepsis 24 h group (p0.05).
4. The level of TNF-alpha in serum was significantly higher in sepsis group than in normal group (p0.05). The level of TNF-alpha in Xuebijing group was significantly lower than that in sepsis group (p0.05). The difference was statistically significant (p0.05). The levels of TNF-alpha in 12-hour sepsis group and 24-hour sepsis group were significantly higher than those in 6-hour sepsis group (p0.05). There was no significant difference in the levels of TNF-alpha between 12-hour sepsis group and 24-hour sepsis group (p0.05).
5. In normal lung tissues, the positive rate of Annexin A1 expression was lower. In sepsis group, the positive rate of Annexin A1 expression was higher than that in normal lung tissues, and there was statistical difference (p0.05). In Xuebijing treatment group (6h, 12h, 24h), the positive rate of Annexin A1 expression was stronger than that in sepsis group (p0.05).
Conclusion: 1. The level of Annexin A1 in the neutrophils and lungs of septic rats increased gradually with the development of the disease and reached a balance between 12 and 24 hours.
2. The levels of TNF-alpha and IL-8 in the serum of septic rats increased gradually with the development of the disease; Xuebijing injection could reduce the levels of TNF-alpha and IL-8 in the serum of septic rats after intervention.
3. Xuebijing injection can effectively improve the condition of rats with Acinetobacter baumannii sepsis, significantly inhibit the excessive release of inflammatory mediators IL-8 and TNF-alpha, and increase the content of Annexin A1 in vivo, but the mechanism of Annexin A1 affecting the level of IL-8 and TNF-alpha in vivo needs further confirmation.
【學(xué)位授予單位】：蚌埠醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R459.7
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