本文選題：短刺法 + 膝骨關(guān)節(jié)炎�。� 參考：《重慶醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的觀察“短刺加電針法”對膝骨關(guān)節(jié)炎模型兔血清中CTX-II、PIICP含量與膝關(guān)節(jié)軟骨中II型膠原(Collagen Type II)—盤狀結(jié)構(gòu)域受體(discoidin domain receptor,DDR)-2—基質(zhì)金屬蛋白酶(matrix metalloproteinase,MMP)-13信號通路的影響,探討該治療方法修復(fù)膝關(guān)節(jié)軟骨細胞外基質(zhì)的可能作用機制。方法將40只新西蘭大白兔隨機分為正常組(10只)和造模組(30只),造模組動物采用Hulth-Telhag法手術(shù)復(fù)制膝骨關(guān)節(jié)炎模型,Lequesne MG膝關(guān)節(jié)級別評估法對各組兔進行評價,X線評估造模結(jié)果。將造模成功的動物隨機分為3組,即模型組、短刺組、普通針刺組,每組10只。短刺組采用短刺加電針法,普通針刺組采用常規(guī)針刺加電針法治療,均選取“內(nèi)膝眼”、“犢鼻”、“陰陵泉”、“足三里”和“梁丘”進行針刺。正常組和模型組正常抓取、固定,不予以干預(yù)。每次治療20min,每日1次,5d為一療程,每個療程間隔2d,共治療4個療程。療程結(jié)束后,Lequesne MG法再次評估各組兔膝關(guān)節(jié)級別,并對各組動物進行膝關(guān)節(jié)半月板及軟骨取材。行HE染色,于光鏡下觀察軟骨細胞的變化;行鉛鈾雙重染色,于透射電鏡下觀察關(guān)節(jié)軟骨超微結(jié)構(gòu)和細胞器結(jié)構(gòu)的變化。采用蛋白免疫印跡法檢測軟骨DDR2、II型膠原、MMP13的蛋白表達;免疫組化法檢測軟骨DDR2、II型膠原、MMP13的表達;逆轉(zhuǎn)錄聚合酶鏈式反應(yīng)檢測DDR2和MMP13的m RNA表達;酶聯(lián)免疫吸附測定法檢測血清PIICP、CTX-II含量。結(jié)果1.X線結(jié)果顯示,造模組兔膝關(guān)節(jié)較正常組改變明顯,造模組兔膝關(guān)節(jié)對線不良,內(nèi)側(cè)關(guān)節(jié)間隙狹窄,關(guān)節(jié)面粗糙變形,關(guān)節(jié)邊緣有骨贅形成。2.光鏡下觀察HE染色結(jié)果顯示,與正常組相比,模型組軟骨細胞排列紊亂,細胞成簇出現(xiàn),細胞核裂解退化明顯;與模型組相比,短刺組和普通針刺組軟骨細胞排列尚齊,成簇存在。透射電鏡檢查結(jié)果顯示,與正常組相比,模型組軟骨細胞核固縮,細胞外基質(zhì)中膠原纖維排列不齊或降解;與模型組相比,短刺組和普通針刺組軟骨細胞核形態(tài)較正常,基質(zhì)中膠原纖維較豐富。3.Lequesne MG評分結(jié)果顯示,造模后6周,模型組、短刺組和普通針刺組兔膝關(guān)節(jié)均出現(xiàn)強烈的局部疼痛反應(yīng),關(guān)節(jié)腫脹明顯且活動度多在15-45度。與正常組相比,模型組、短刺組和普通針刺組Lequesne MG評分均顯著增高(p0.01);造模后10周,短刺組和普通針刺組兔膝關(guān)節(jié)局部疼痛刺激反應(yīng)、膝關(guān)節(jié)腫脹程度均減輕,活動度可達45-90度。與造模后6周同組別兔Lequesne MG評分相比均明顯降低(p0.01)。4.Western-Blot、免疫組化、RT-PCR、ELISA結(jié)果顯示,與正常組相比,模型組、短刺組和普通針刺組兔血清中PIICP含量與膝關(guān)節(jié)軟骨中II型膠原的蛋白表達等均明顯減少(均p0.01),而血清中CTX-II含量與膝關(guān)節(jié)軟骨中DDR2、MMP13的蛋白及m RNA表達均明顯增加(均p0.01);治療后,短刺組和普通針刺組兔血清中PIICP含量與膝關(guān)節(jié)軟骨中II型膠原的蛋白表達等較模型組明顯增多(均p0.01),而血清中CTX-II含量與膝關(guān)節(jié)軟骨中DDR2、MMP13的蛋白及m RNA表達較模型組均明顯減少(均p0.01);與普通針刺組相比,短刺組兔血清中PIICP含量與膝關(guān)節(jié)軟骨中II型膠原的蛋白表達增多(p0.01),而血清中CTX-II含量與膝關(guān)節(jié)軟骨中DDR2、MMP13的蛋白及m RNA表達減少(均p0.01)。結(jié)論本次研究采用Hulth-Telhag法成功復(fù)制了膝骨關(guān)節(jié)炎兔模型。普通電針法和短刺加電針法均能調(diào)整軟骨細胞排列方式,抑制細胞外基質(zhì)的異常降解,恢復(fù)細胞外基質(zhì)的合成,促進兔膝關(guān)節(jié)軟骨修復(fù);且短刺加電針法較普通電針法有一定的療效優(yōu)勢。其作用機制可能與其阻斷II型膠原/DDR2/MMP13通路的病理循環(huán)有關(guān)。
[Abstract]:Objective To observe the effect of "short prickly plus Electroacupuncture" on CTX-II, PIICP content in the serum of knee osteoarthritis model rabbits and the effect of II collagen (Collagen Type II) in the cartilage of knee joint (discoidin domain receptor, DDR) -2 matrix metalloproteinase. The possible mechanism of repairing the cartilage extracellular matrix of the knee joint. Methods 40 New Zealand white rabbits were randomly divided into the normal group (10) and the model group (30). The model animal model was replicated by Hulth-Telhag method. The Lequesne MG knee assessment method was used to evaluate the rabbits in each group. The results of the X-ray evaluation were made. The results of the model were made. The results of the model were made. The animals were randomly divided into 3 groups, namely, model group, short stab group, common acupuncture group, 10 in each group. Short thorn group was treated with short stab plus Electroacupuncture, and ordinary acupuncture group was treated with conventional acupuncture plus Electroacupuncture. All of them selected "inner knee eye", "calf nose", "Yin Ling spring", "foot three li" and "Liang Qiu" by acupuncture. Normal group and model group were normal grasp. After the treatment of 20min, 1 times a day, 5D was a course of treatment, each course was 2D, and 4 courses were treated. After the end of the course, the knee joint level of each group was evaluated by Lequesne MG method again, and the meniscus and cartilage of the knee joint were taken from each group. The changes of cartilage cells were observed under the light microscope, and the lead and uranium were observed under the light microscope. Double staining was used to observe the changes of ultrastructure and organelle structure of articular cartilage under transmission electron microscope. The protein expression of cartilage DDR2, type II collagen and MMP13 was detected by protein immunoblotting; immunohistochemical method was used to detect the expression of cartilage DDR2, II collagen, MMP13; reverse transcription polymerase chain reaction was used to detect the m RNA expression of DDR2 and MMP13; enzyme linked immunosorbent assay The results of 1. X-ray results showed that the knee joint of the model rabbit model was more obvious than that of the normal group, the knee joint of the model rabbit model was bad, the medial joint space was narrow, the joint surface was rough, and the joint edge had the osteophyte formation under the.2. light microscope, and the HE staining results showed that the model group was soft compared with the normal group. Compared with the model group, the cartilage cells in the short stings group and the common acupuncture group were arranged and clustered. The results of transmission electron microscopy showed that compared with the normal group, the nuclei of the cartilage cells in the model group were fixed and the collagen fibers in the extracellular matrix were inhomogeneous or degraded; and the model group was the same as the model group. Compared with the short prickle group and the common acupuncture group, the morphology of the cartilage nuclei was more normal. The.3.Lequesne MG score of the collagen fiber in the matrix showed that the knee joint of the model group, the short spiny group and the common acupuncture group had strong local pain response 6 weeks after the model, and the joint swelling was obvious and the activity was 15-45 degrees more. Compared with the normal group, the model was compared with the normal group. The score of Lequesne MG in the short spines group and the common acupuncture group increased significantly (P0.01), and the knee joint pain stimulation reaction in the short stings group and the common acupuncture group after 10 weeks of modeling was reduced and the activity of the knee joint was 45-90 degrees. Compared with the Lequesne MG score of the group of the rabbits in the same group at 6 weeks after making the model, the.4.Western-Blot was significantly reduced (P0.01).4.Western-Blot. The results of phytophthoraczation, RT-PCR and ELISA showed that compared with the normal group, the content of PIICP in the serum of the rabbits in the model group, the short spiny group and the common acupuncture group and the expression of the protein expression of type II collagen in the cartilage of the knee were significantly decreased (P0.01), but the content of CTX-II in the serum and the expression of DDR2, MMP13 and m RNA in the cartilage of the knee joint were significantly increased (P0.01). After the treatment, the content of PIICP in the serum and the protein expression of type II collagen in the articular cartilage of the short spiny group and the common acupuncture group increased significantly (P0.01), while the CTX-II content in the serum and the expression of DDR2, MMP13 and m RNA in the cartilage of the knee joint were significantly less than those in the model group (all P0.01). Compared with the common acupuncture group, the blood of the short spiny group was compared with the normal acupuncture group. The content of PIICP and the protein expression of type II collagen in the cartilage of the knee joint increased (P0.01), while the CTX-II content in the serum and the expression of DDR2, MMP13 and m RNA in the cartilage of the knee joint decreased (P0.01). Conclusion the rabbit model of knee osteoarthritis was successfully replicated by Hulth-Telhag method. The common electric needle method and the short spiny acupuncture and electroacupuncture were all adjustable. The arrangement of the whole cartilage cells can inhibit the abnormal degradation of the extracellular matrix, restore the synthesis of extracellular matrix and promote the repair of the cartilage of the knee joint of rabbits, and the short spiny and electroacupuncture method has a certain therapeutic advantage over the common electroacupuncture method. Its mechanism may be related to the pathological cycle of blocking the II collagen /DDR2/MMP13 pathway.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R245;R-332

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