本文選題：硫酸軟骨素 + 低分子量�。� 參考：《青島大學(xué)》2014年碩士論文

【摘要】：目的 研究低分子硫酸軟骨素的制備方法,并探討其對(duì)軟骨的修復(fù)作用。 方法 1、稱取一定量分子質(zhì)量在5萬(wàn)以上的硫酸軟骨素(ChS),在一定溫度和條件下分別用HC1、H2O2和硫酸軟骨素裂解酶EAC對(duì)其進(jìn)行降解,反應(yīng)結(jié)束后用乙醇沉淀、脫水、干燥,產(chǎn)物用高效液相色譜儀測(cè)定分子量。 2、建立新西蘭大白兔膝關(guān)節(jié)軟骨缺失模型,36只新西蘭大白兔隨機(jī)分成6組,分別為對(duì)照組[蒸餾水0.4g/(kg·d)]、模型組[蒸餾水0.4g/(kg·d)]、低分子量ChS低劑量組[低分子硫酸軟骨素0.2g/(kg·d)]、低分子量ChS高劑量組[低分子硫酸軟骨素0.4g/(kg·d)]、高分子量ChS低劑量組[高分子硫酸軟骨素0.2g/(kg·d)]和高分子量ChS高劑量組[高分子硫酸軟骨素0.4g/(kg·d)],每組6只,分別進(jìn)行灌胃給藥。記錄各組實(shí)驗(yàn)用兔的體重、生存時(shí)間和生活狀況,對(duì)關(guān)節(jié)軟骨缺損部位修復(fù)情況和病理組織學(xué)切片進(jìn)行觀察。應(yīng)用ELISA法檢測(cè)兔關(guān)節(jié)液中IL-1β、TNF-β和TGF-β的水平,并采用全自動(dòng)生化分析儀及其配套的試劑盒對(duì)兔血清中鈣、磷含量進(jìn)行測(cè)定。 結(jié)果 1、經(jīng)高效液相色譜法測(cè)定,硫酸軟骨素原料的分子量為55426。用HC1降解得到的ChS,分子量為3936,顏色從白色轉(zhuǎn)變成紅褐色,產(chǎn)物粘性較大,產(chǎn)率較低為48.71%；而用H202在酸性條件下和用硫酸軟骨素裂解酶EAC降解所得到的硫酸軟骨素,降解效果較好,分子量分別為4217和4565,其外觀較好,產(chǎn)物為乳白色,其產(chǎn)率分別達(dá)到64.45%和91.6%。 2、造模后傷口愈合情況較好,實(shí)驗(yàn)用兔生活狀態(tài)和精神狀態(tài)良好。造模后各組體重均有下降,但模型組與其余給予ChS組相比,體重下降明顯。低分子ChS組,其軟骨表面較為平整,凹陷基本已填充,較模型組與高分子ChS組效果明顯。在HE染色狀態(tài)下,損傷修復(fù)處均可見(jiàn)初期修復(fù)反應(yīng),并且給予低分子ChS組效果要優(yōu)于高分子ChS組和模型組。 3、低分子ChS和高分子ChS都能夠減少關(guān)節(jié)液中炎性因子的含量和增加TGF-β的含量(P0.05),且低分子ChS組與高分子ChS組相比,效果較好(P0.05)。與模型組相比,給予ChS的組別,其血清中的鈣磷含量也相對(duì)較少(P0.05)。 結(jié)論 1、HC1、H2O2和硫酸軟骨素裂解酶EAC都能有效地將分子量為5萬(wàn)以上的ChS降解為分子量在3500-5300范圍內(nèi)的低分子量產(chǎn)物。 2、低分子ChS能夠更好地促進(jìn)傷口愈合和軟骨修復(fù),這可能是因?yàn)榈头肿覥hS,分子量較小,機(jī)體易于吸收,生物利用度提高,從而使得傷口愈合和軟骨修復(fù)的作用較為明顯。 3、高、低分子量的ChS都可以增加TGF-β的含量,降低IL-1β、TNF-α和血清中鈣、磷的含量。且低分子ChS要優(yōu)于高分子ChS。對(duì)于軟骨修復(fù),這可能是通過(guò)對(duì)上述因子的影響,從而產(chǎn)生積極的作用。
[Abstract]:Purpose To study the preparation of chondroitin sulfate and its effect on cartilage repair. Method 1. Chondroitin sulfate chondroitin sulfate (CHS) with a molecular weight of more than 50 000 was obtained and degraded by HC1H 2O 2 and chondroitin sulfate lyase EAC at certain temperature and under certain conditions. After the reaction was finished, the chondroitin sulfate was precipitated, dehydrated and dried. The molecular weight of the product was determined by high performance liquid chromatography. 2, 36 New Zealand white rabbits were randomly divided into 6 groups. Control group [distilled water 0.4g/(kg d], model group [distilled water 0.4g/(kg d], low molecular weight ChS low dose group [low molecular weight chondroitin sulfate 0.2g/(kg d], low molecular weight ChS high dose group [low molecular weight chondroitin sulfate 0.4g/(kg d], high molecular weight ChS low dose group [high molecular weight ChS low dose group] Chondroitin sulfate (0.2g/(kg d) and high molecular weight ChS (high dose group [high molecular weight chondroitin sulfate 0.4g/(kg d]), 6 rats in each group, The drugs were administered by gavage respectively. The weight, survival time and living condition of the rabbits were recorded. The repair of articular cartilage defects and histopathological sections were observed. The levels of IL-1 尾 -TNF- 尾 and TGF- 尾 in rabbit articular fluid were detected by ELISA method. The contents of calcium and phosphorus in rabbit serum were determined by automatic biochemical analyzer and kit. Result 1. The molecular weight of chondroitin sulfate was 55426 by HPLC. Chondroitin sulfate obtained by HC1 degradation, with a molecular weight of 3936, was changed from white to reddish brown, with a high viscosity and a lower yield of 48.71.The degradation of chondroitin sulfate by H202 under acidic conditions and by chondroitin sulfate lyase EAC showed better degradation effect. The molecular weight was 4217 and 4565 respectively. The product was milky white and its yield reached 64.45% and 91.6% respectively. 2. The wound healed well after modeling, and the living state and mental state of experimental rabbits were good. The weight of each group was decreased after the model was made, but the weight of the model group was significantly lower than that of the other groups treated with ChS. In the low molecular weight ChS group, the cartilage surface was flat and the hollow was basically filled, which was more effective than the model group and the high molecular weight ChS group. Under HE staining, the initial repair reaction was observed at the site of injury repair, and the effect of low molecular weight ChS group was better than that of macromolecule ChS group and model group. 3. Both low molecular weight ChS and high molecular weight ChS could decrease the content of inflammatory factor and increase the content of TGF- 尾 in articular fluid. The effect of low molecular weight ChS group was better than that of high molecular weight ChS group. Compared with the model group, the serum calcium and phosphorus contents of the ChS group were also relatively lower than that of the model group. Conclusion 1HC1H _ 2O _ 2 and chondroitin sulfate lyase EAC can effectively degrade ChS with molecular weight over 50,000 to low molecular weight products in the range of 3500-5300. 2, low molecular weight ChS can better promote wound healing and cartilage repair, which may be due to the low molecular weight, easy to be absorbed and bioavailability, which makes wound healing and cartilage repair more obvious. 3. ChS with high, low molecular weight could increase the content of TGF- 尾, decrease the content of IL-1 尾 TNF- 偽 and the contents of calcium and phosphorus in serum. And the low molecular ChS is superior to the polymer Chs. For cartilage repair, this may have a positive effect by influencing these factors.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R914

【引證文獻(xiàn)】

相關(guān)期刊論文 前1條

1 王俊;王紫薇;張金木;汪蘭;喬宇;李新;扥曉艷;吳文錦;熊光權(quán);;不同來(lái)源和純度的硫酸軟骨素理化特性的研究[J];湖北農(nóng)業(yè)科學(xué);2016年23期

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本文編號(hào)：1798087