痹祺提取物對CIA大鼠滑膜增殖及軟骨破壞的影響
發(fā)布時間：2018-10-20 09:06
【摘要】：目的:類風(fēng)濕關(guān)節(jié)炎(rheumatoid arthritis,RA)以關(guān)節(jié)腫痛及進行性破壞為主要臨床表現(xiàn),是一種長期的、慢性的、全身性的自身免疫性疾病,該病女性多發(fā),且30-50歲為高發(fā)年齡,中國大陸的患病率為0.2%-4%。目前針對RA的纖維樣滑膜細(xì)胞(Fibroblast like synoviocytes,FLS)的增生、轉(zhuǎn)化及侵襲、滑膜新生血管增殖、血管翳的形成、軟骨的降解與破壞、骨質(zhì)侵蝕等病理方面的靶向性治療為當(dāng)前研究的重點與難點。痹祺提取物為目前臨床治療RA應(yīng)用較多的藥物,但其對RA的藥理作用機制的研究尚不完整,故本實驗以CIA大鼠為動物模型,模擬RA發(fā)病過程,觀察痹祺提取物對CIA大鼠TNF-α、IL-18、OPN、COMP的血清水平及OPN、COMP分別在軟骨、滑膜表達(dá)的影響,初步探討痹祺膠囊干預(yù)CIA大鼠關(guān)節(jié)炎癥、滑膜增生及軟骨破壞的藥理作用機制,為臨床上治療RA提供可靠的實驗證據(jù)。方法:[1]建立CIA大鼠模型,觀察并計算大鼠足腫脹率。[2]采用ELISA檢測大鼠造模2W時血清TNF-α及灌藥4W后血清TNF-α、IL-18、OPN、COMP水平。[3]采用IHC檢測OPN、COMP在滑膜中的表達(dá)。[4]采用RT-PCR檢測OPN、COMP在軟骨中的表達(dá)。[5]顯微鏡下觀察HE染色滑膜組織、軟骨組織病理形態(tài)。結(jié)果:[1]成功建立CIA模型,CIA模型組較正常組大鼠足腫脹率明顯升高,治療組較模型組大鼠足腫脹率明顯減低(P0.05)。[2]細(xì)胞因子血清水平比較:CIA模型組較正常組大鼠治療后TNF-α、IL-18、OPN、COMP在血清中表達(dá)量升高(P0.05),治療組較模型組表達(dá)量降低(P0.05)。[3]滑膜組織中細(xì)胞因子的蛋白表達(dá):正常組滑膜區(qū)可見OPN少量表達(dá),正常組滑膜區(qū)未見COMP的表達(dá),模型組滑膜區(qū)可見大量陽性細(xì)胞,模型組較正常組陽性細(xì)胞增多(P0.05),治療組滑膜區(qū)可見部分陽性細(xì)胞,較模型組減少(P0.05)。[4]軟骨中細(xì)胞因子的蛋白表達(dá):實時相對定量分析結(jié)果顯示大鼠軟骨OPN mRNA、COMP mRNA表達(dá)模型組較正常組升高(P0.05),治療組較模型組降低(P0.05)。[5]HE染色病理圖片觀察:參與滑膜組織病理評分計算滑膜炎癥程度、軟骨病理評分計算軟骨損傷程度,模型組較正常組病理評分升高(P0.05),治療組較模型組病理評分下降(P0.05),中劑量組在炎性細(xì)胞浸潤及滑膜細(xì)胞增生及軟骨損傷方面較其他治療組降低(P0.05)。結(jié)論:[1]痹祺提取物干預(yù)大鼠TNF-α、IL-18在血清中的表達(dá),阻斷炎性因子網(wǎng)絡(luò)形成,從而干預(yù)CIA關(guān)節(jié)炎癥的進一步發(fā)展。[2]痹祺提取物可能通過下調(diào)大鼠OPN在血清中及OPN、COMP在滑膜中的表達(dá),干預(yù)滑膜腫瘤樣增生,從而抑制滑膜血管新生及血管翳的形成。[3]痹祺提取物可能通過下調(diào)大鼠OPN、COMP在軟骨中的表達(dá),干預(yù)軟骨基質(zhì)降解,從而阻止軟骨的破壞。[4]大鼠COMP血清水平模型組較正常組增加,反映CIA大鼠出現(xiàn)軟骨破壞,經(jīng)過痹祺提取物治療的大鼠COMP血清水平較模型組下降,反映其治療CIA大鼠軟骨破壞有效。
[Abstract]:Objective: rheumatoid arthritis (rheumatoid arthritis,RA) is a long-term, chronic, systemic autoimmune disease with the main clinical manifestation of joint swelling and pain and progressive destruction. The prevalence rate in mainland China was 0.2-4. The proliferation, transformation and invasion of RA fibroid synovial cells (Fibroblast like synoviocytes,FLS), the proliferation of synovial neovascularization, the formation of pannus, the degradation and destruction of cartilage, Targeted treatment of bone erosion and other pathological aspects is the focus and difficulty of current research. Biqi extract is a widely used drug in clinical treatment of RA at present, but its pharmacological mechanism to RA is not complete. Therefore, CIA rats are used as animal models in this experiment to simulate the pathogenesis of RA. To observe the effects of Biqi extract on the serum levels of TNF- 偽 and IL-18,OPN,COMP and the expression of OPN,COMP in cartilage and synovium of CIA rats, and to explore the pharmacological mechanism of Biqi capsule in preventing arthritis, synovial hyperplasia and cartilage destruction in CIA rats. To provide reliable experimental evidence for clinical treatment of RA. Methods: [1] CIA rat model was established. The rate of rat foot swelling was observed and calculated. [2] the levels of serum TNF- 偽 and IL-18,OPN,COMP were detected by ELISA at 2 W and 4 W after administration. [3] IHC was used to detect the expression of OPN,COMP in synovium. [4] RT-PCR was used to detect the expression of OPN,COMP in cartilage. [5] Microscopy was used to detect the expression of OPN,COMP. HE staining of synovial tissue, Cartilage histopathology. Results: [1] CIA model was successfully established, and the rate of foot swelling in the CIA model group was significantly higher than that in the normal group. (2) comparison of serum levels of cytokines: the expression of TNF- 偽 and IL-18,OPN,COMP in serum of CIA model group was higher than that of normal group (P0.05), and that of treatment group was lower than that of model group (P0.05). [3] in synovial tissue, the expression of TNF- 偽 and IL-18,OPN,COMP in the treatment group was significantly lower than that in the model group (P0.05). Protein expression of cytokines: in normal group, a small amount of OPN was expressed in synovial region. The expression of COMP was not found in the synovial region of the normal group. A large number of positive cells were found in the synovial area of the model group, and the number of positive cells in the model group was higher than that in the normal group (P0.05). Some positive cells were found in the synovial area of the treatment group. Compared with the model group (P0.05). [4] the expression of cytokines in cartilage: the results of real-time relative quantitative analysis showed that the expression of OPN mRNA,COMP mRNA in the model group was higher than that in the normal group (P0.05), and that in the treatment group was lower than that in the model group (P0.05). [5] the pathological pictures of HE staining were observed. The degree of synovitis was calculated by participating in synovial tissue pathological score. The degree of cartilage injury in model group was higher than that in normal group (P0.05), the pathological score in treatment group was lower than that in model group (P0.05), and the inflammatory cell infiltration, synovial cell proliferation and cartilage injury in middle dose group were lower than that in other treatment group (P0.05). Conclusion: [1] Biqi extract interferes with the expression of TNF- 偽 and IL-18 in serum of rats, blocks the formation of inflammatory factor network, and interferes with the further development of CIA arthritis. [2] Biqi extract may down-regulate the expression of OPN in serum and OPN,COMP in synovium of rats. Intervention of synovial tumor-like hyperplasia, thereby inhibiting the formation of synovial angiogenesis and pannus. [3] Biqi extract may interfere with cartilage matrix degradation by down-regulating the expression of OPN,COMP in cartilage. [4] the serum level of COMP in the model group was higher than that in the normal group, reflecting cartilage destruction in the CIA rats, and the serum level of COMP in the rats treated with Biqi extract was lower than that in the model group. It is effective in the treatment of cartilage destruction in CIA rats.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R593.22
本文編號：2282688
[Abstract]:Objective: rheumatoid arthritis (rheumatoid arthritis,RA) is a long-term, chronic, systemic autoimmune disease with the main clinical manifestation of joint swelling and pain and progressive destruction. The prevalence rate in mainland China was 0.2-4. The proliferation, transformation and invasion of RA fibroid synovial cells (Fibroblast like synoviocytes,FLS), the proliferation of synovial neovascularization, the formation of pannus, the degradation and destruction of cartilage, Targeted treatment of bone erosion and other pathological aspects is the focus and difficulty of current research. Biqi extract is a widely used drug in clinical treatment of RA at present, but its pharmacological mechanism to RA is not complete. Therefore, CIA rats are used as animal models in this experiment to simulate the pathogenesis of RA. To observe the effects of Biqi extract on the serum levels of TNF- 偽 and IL-18,OPN,COMP and the expression of OPN,COMP in cartilage and synovium of CIA rats, and to explore the pharmacological mechanism of Biqi capsule in preventing arthritis, synovial hyperplasia and cartilage destruction in CIA rats. To provide reliable experimental evidence for clinical treatment of RA. Methods: [1] CIA rat model was established. The rate of rat foot swelling was observed and calculated. [2] the levels of serum TNF- 偽 and IL-18,OPN,COMP were detected by ELISA at 2 W and 4 W after administration. [3] IHC was used to detect the expression of OPN,COMP in synovium. [4] RT-PCR was used to detect the expression of OPN,COMP in cartilage. [5] Microscopy was used to detect the expression of OPN,COMP. HE staining of synovial tissue, Cartilage histopathology. Results: [1] CIA model was successfully established, and the rate of foot swelling in the CIA model group was significantly higher than that in the normal group. (2) comparison of serum levels of cytokines: the expression of TNF- 偽 and IL-18,OPN,COMP in serum of CIA model group was higher than that of normal group (P0.05), and that of treatment group was lower than that of model group (P0.05). [3] in synovial tissue, the expression of TNF- 偽 and IL-18,OPN,COMP in the treatment group was significantly lower than that in the model group (P0.05). Protein expression of cytokines: in normal group, a small amount of OPN was expressed in synovial region. The expression of COMP was not found in the synovial region of the normal group. A large number of positive cells were found in the synovial area of the model group, and the number of positive cells in the model group was higher than that in the normal group (P0.05). Some positive cells were found in the synovial area of the treatment group. Compared with the model group (P0.05). [4] the expression of cytokines in cartilage: the results of real-time relative quantitative analysis showed that the expression of OPN mRNA,COMP mRNA in the model group was higher than that in the normal group (P0.05), and that in the treatment group was lower than that in the model group (P0.05). [5] the pathological pictures of HE staining were observed. The degree of synovitis was calculated by participating in synovial tissue pathological score. The degree of cartilage injury in model group was higher than that in normal group (P0.05), the pathological score in treatment group was lower than that in model group (P0.05), and the inflammatory cell infiltration, synovial cell proliferation and cartilage injury in middle dose group were lower than that in other treatment group (P0.05). Conclusion: [1] Biqi extract interferes with the expression of TNF- 偽 and IL-18 in serum of rats, blocks the formation of inflammatory factor network, and interferes with the further development of CIA arthritis. [2] Biqi extract may down-regulate the expression of OPN in serum and OPN,COMP in synovium of rats. Intervention of synovial tumor-like hyperplasia, thereby inhibiting the formation of synovial angiogenesis and pannus. [3] Biqi extract may interfere with cartilage matrix degradation by down-regulating the expression of OPN,COMP in cartilage. [4] the serum level of COMP in the model group was higher than that in the normal group, reflecting cartilage destruction in the CIA rats, and the serum level of COMP in the rats treated with Biqi extract was lower than that in the model group. It is effective in the treatment of cartilage destruction in CIA rats.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R593.22
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