本文選題：維生素D3 + 脂質(zhì)體　； 參考：《吉林大學(xué)》2017年碩士論文

【摘要】：維生素D3(Vitamin D3,VD3)是一種脂溶性的維生素,屬于一種作用于鈣磷代謝的激素前體,又叫做“陽(yáng)光維生素”。VD3具有很多方面的功效,如調(diào)節(jié)鈣磷代謝,預(yù)防骨質(zhì)疏松,改善心腦血管疾病,提高免疫力等作用。但是由于VD3是脂溶性的,不溶于水,限制了其藥效。為了提高VD3的生物利用度,需要一種更為安全高效的VD3制劑。脂質(zhì)體是一種單層或多層兩性雙分子層組成的結(jié)構(gòu),內(nèi)部包含水溶性或脂溶性物質(zhì),可以作為藥物載體,達(dá)到提高藥效和降低副作用的目的。而本文主要利用VD3的高效抗氧化性,阻止皮膚細(xì)胞中自由基的過(guò)度產(chǎn)生,提高自由基清除酶的活性,將VD3與脂質(zhì)體的結(jié)合,制備成維生素D3脂質(zhì)體(L-VD3)來(lái)達(dá)到防止皮膚光老化的作用。本文主要包括如下內(nèi)容:1.VD3體外分析方法學(xué)的建立。建立了VD3的體外分析方法-高效液相色譜法(HPLC),使用此方法得到的VD3的精密度和回收率均符合體外分析測(cè)定要求。HPLC測(cè)定L-VD3中的藥物含量,并且該方法測(cè)定的L-VD3中藥物的回收率符合體外分析要求。建立了用超濾離心法來(lái)分離游離VD3,進(jìn)而測(cè)定L-VD3的包封率的方法。此方法可以很好的使游離藥物分離,并且方便有效,且該法測(cè)定的VD3的精密度和回收率也均符合體外分析測(cè)定的要求。2.VD3常規(guī)脂質(zhì)體制備及性能評(píng)價(jià)分別就乙醇注入法,薄膜均質(zhì)法,以及反向蒸發(fā)法對(duì)L-VD3的制備方法進(jìn)行了篩選,對(duì)比包封率和脂質(zhì)體的粒徑,得到乙醇注入法是制備L-VD3的最適合的方法。接著分別就L-VD3配方中磷脂含量、磷脂與膽固醇的比例、脂藥比以及磷酸緩沖液的PH值進(jìn)行單因素實(shí)驗(yàn)。篩選出來(lái)磷脂含量、磷脂與膽固醇的比例、脂藥比作為主要的考察對(duì)象進(jìn)行響應(yīng)面優(yōu)化實(shí)驗(yàn),得到最優(yōu)工藝配方為:磷脂含量為25mg,磷脂與膽固醇的比例為4.5:1,脂藥比為2.5:l。以最優(yōu)配方條件下得到vd3常規(guī)脂質(zhì)體的包封率的理論值為87.83%,實(shí)際值為86.79%。3.l-vd3的穩(wěn)定性研究本文在溫度、光照、凍融的條件下進(jìn)行了影響l-vd3穩(wěn)定性因素的測(cè)試。其中溫度在4-40℃的條件下,包封率基本不發(fā)生太大變化,穩(wěn)定性較好,適宜保存l-vd3;光照對(duì)l-vd3的穩(wěn)定性沒有過(guò)多影響,因此可以說(shuō)明vd3在制備成脂質(zhì)體之后對(duì)光的穩(wěn)定性大大提高了,可以不用避光保存;凍融條件對(duì)l-vd3的穩(wěn)定性也沒有過(guò)多影響,可以說(shuō)明vd3制成脂質(zhì)體后對(duì)溫度的突變穩(wěn)定性較好,也可以很好的適應(yīng)溫度的變化。另外,本文進(jìn)行了vd3的體外釋放試驗(yàn),發(fā)現(xiàn)l-vd3在皮膚中的單位截留藥量為182.13μg/cm2,而vd3溶液在皮膚中的單位截留藥量為113.82μg/cm2,l-vd3的單位截留藥量要遠(yuǎn)大于vd3溶液,并且l-vd3的透皮量也小于vd3溶液,因此將vd3做成脂質(zhì)體會(huì)更有利于皮膚吸收。之后又進(jìn)行了加速實(shí)驗(yàn)和長(zhǎng)期實(shí)驗(yàn),發(fā)現(xiàn)l-vd3的包封率和其外觀形態(tài)并沒有太大的變化,說(shuō)明l-vd3的穩(wěn)定性較好。4.l-vd3的藥效研究本文建立了皮膚光老化模型。經(jīng)過(guò)曬傷處理的大鼠皮膚都有明顯的褶皺,紅腫,斑點(diǎn),白塊等不同程度的光老化現(xiàn)象,說(shuō)明模型建立成功。藥物治療了一段時(shí)間之后觀察大鼠的皮膚狀態(tài),發(fā)現(xiàn)涂抹l-vd3和陽(yáng)性對(duì)照藥物后的大鼠皮膚基本上恢復(fù)正常,沒有明顯的白斑褶皺破損等現(xiàn)象,光潔有彈性;涂抹了vd3溶液的大鼠皮膚狀況也有一定的改善,雖然仍有有一些黃斑紅點(diǎn),但不是很多;而涂抹pbs溶液的陰性對(duì)照組大鼠皮膚依舊有紅斑出血白斑褶皺等光老化現(xiàn)象,沒有得到改善。之后將各組大鼠的皮膚進(jìn)行he染色,觀察切片的電鏡圖片,發(fā)現(xiàn)涂抹l-vd3和陽(yáng)性對(duì)照藥物的大鼠表皮真皮結(jié)構(gòu)基本完整,內(nèi)部纖維組織、腺體等排布均勻,與正常的大鼠皮膚組織切片狀態(tài)相差無(wú)幾;涂抹了vd3溶液的大鼠表皮真皮結(jié)構(gòu)略有改變,內(nèi)部纖維組織、腺體個(gè)別變形,但整體上比較有序排列;而涂抹pbs溶液的陰性對(duì)照組大鼠表皮真皮嚴(yán)重分層,之間的界限模糊不清,內(nèi)部腺體纖維也雜亂無(wú)章甚至變形,皮膚損傷狀況沒有改善。以上結(jié)果說(shuō)明,L-VD3可以有效的防止皮膚的光老化現(xiàn)象,有比較顯著的效果。
[Abstract]:Vitamin D3 (Vitamin D3, VD3) is a fat soluble vitamin. It belongs to a hormone precursor that acts on calcium and phosphorus metabolism. It is also called "sunshine vitamin".VD3, which has many effects, such as regulating calcium and phosphorus metabolism, preventing osteoporosis, improving cardiovascular and cerebrovascular diseases, and improving immunity. But because VD3 is fat soluble, insoluble. In order to improve the bioavailability of VD3, in order to improve the bioavailability of the VD3, a more safe and efficient preparation is needed. The liposome is a structure composed of a single layer or multilayer bisexual biolecular layer, which contains water-soluble or fat soluble substances, which can be used as a drug carrier to achieve high efficacy and reduce side effects. The effective antioxidant activity of VD3 is used to prevent the excessive production of free radicals in skin cells, improve the activity of free radical scavenging enzymes, and combine VD3 with liposomes to prepare vitamin D3 liposomes (L-VD3) to prevent skin photoaging. This article mainly includes the following contents: the establishment of the method of 1.VD3 in vitro analysis and the establishment of the body of VD3. The external analysis method - high performance liquid chromatography (HPLC), the precision and recovery of VD3 obtained by this method are all in accordance with the requirement for the determination of the drug content in L-VD3 by.HPLC in vitro, and the recovery rate of the drug in L-VD3 is in conformity with the requirements of the in vitro analysis. The separation of free VD3 is established by ultrafiltration centrifugation, and the determination of L-V is established. The method of encapsulation rate of D3. This method can make the separation of free drugs very good, and it is convenient and effective, and the precision and recovery of VD3 measured by this method also conform to the requirements of in vitro analysis and determination of.2.VD3. The preparation and performance evaluation of the conventional lipid system and performance evaluation on the ethanol injection method, the thin film homogenization method, and the reverse evaporation method for the preparation of L-VD3. The method was screened to compare the encapsulation efficiency and the particle size of liposomes. Ethanol injection was the most suitable method for preparing L-VD3. Then, the content of phospholipid, the proportion of phospholipid and cholesterol, the ratio of lipid to cholesterol, the pH of the phosphate buffer and the pH value of the phosphoric acid buffer were tested. The content of phospholipid, the proportion of phospholipid and cholesterol, the proportion of lipids were screened, and the proportion of phospholipid and cholesterol was screened. The response surface optimization experiment was carried out as the main object. The optimum formula was obtained: the content of phospholipid was 25mg, the proportion of phospholipid and cholesterol was 4.5:1, and the theoretical value of the encapsulation efficiency of VD3 conventional liposomes was 87.83% under the optimal formulation condition, and the stability of 86.79%.3.l-vd3 was studied in this paper. In the conditions of temperature, light and freezing and thawing, the stability factors of l-vd3 were tested. Under the condition of 4-40 C, the encapsulation efficiency did not change much, the stability was better, and the stability of the l-vd3 was better. The light has not too much effect on the stability of l-vd3, so it can explain the stability of VD3 to the light after preparing the liposome. As a result, the freezing and thawing conditions have no effect on the stability of l-vd3, which shows that the VD3 liposomes have a better temperature mutation and can adapt to the change of temperature. In addition, the release test of VD3 in vitro is carried out in this paper, and the unit interception of l-vd3 in the skin is 182.13. Mu g/cm2, and the unit of VD3 solution in the skin of unit interception of the amount of 113.82 u g/cm2, l-vd3 unit to intercept the dose is much larger than the VD3 solution, and l-vd3 transdermal volume is less than the VD3 solution, so the VD3 to make lipid experience is more beneficial to the skin absorption. Then, the accelerated experiment and long-term experiments were carried out, and the encapsulation rate and external appearance of l-vd3 were found. The state has not changed too much. It shows that the stability of l-vd3 is better than that of.4.l-vd3. The skin photoaging model has been established in this paper. The skin of rats treated with sunburn has obvious folds, redness, blotches, white patches and other light aging phenomena, indicating the success of the model. After a period of drug treatment, the rats were observed. The skin status, the skin of rats after smearing l-vd3 and positive control drugs was basically restored to normal, without obvious leukoplakia fold damage and other phenomena, smooth and flexible; the skin condition of rats smeared with VD3 solution was also improved, although there were some yellow spot red spots, but not many; and the negative control group smeared with PBS solution The skin of rats was still characterized by erythema hemorrhage and leukoplakia folds, which were not improved. Then the skin of rats in each group was stained with he, and the electron microscope pictures of the slices were observed. It was found that the dermal structure of the skin of rats smeared with l-vd3 and positive control drugs was basically complete, the internal fibrous tissue, glands, etc. were evenly distributed, with the normal rat skin. The tissue section of the tissue was different. The dermal structure of the rat epidermis with VD3 solution was slightly changed, the internal fibrous tissue and the gland were deformed, but on the whole, the epidermis of the negative control group smeared with PBS solution was seriously stratified, the boundary between the epidermis was not clear, the inner gland fibers were disorderly and even deformed. The results showed that L-VD3 could effectively prevent photoaging of skin and had a significant effect.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R943;R96

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1 夏鴻熙;VD3脂質(zhì)體的制備及其體內(nèi)外抑制光老化活性初步研究[D];吉林大學(xué);2017年

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本文編號(hào)：2017115