本文選題：胡黃連苷Ⅱ + 脂多糖（LPS）　； 參考：《大連醫(yī)科大學(xué)》2013年碩士論文

【摘要】：目的:葡萄膜炎是常見眼病，也是主要致盲眼病之一，其中以前葡萄膜炎為最多，其次是全葡萄膜炎。在發(fā)達(dá)國家，5-20%的患者由于葡萄膜炎致盲，而在發(fā)展中國家這個比例可能要超過25%，因?yàn)榘l(fā)展中國家影響眼部的傳染病機(jī)會較多，如弓形蟲病和結(jié)核。但是目前葡萄膜炎的治療方法主要是使用激素，激素治療副作用大，眼科方面可能導(dǎo)致激素源性角膜炎、青光眼、白內(nèi)障；全身性副作用包括誘發(fā)高血壓、糖尿病、骨質(zhì)疏松、消化道出血、癲癇發(fā)作等，，這些都導(dǎo)致激素在應(yīng)用過程中具有相當(dāng)?shù)木窒扌�。所以我們尋找一種可以有效治療葡萄膜炎并且毒副作用小的藥物，中國傳統(tǒng)醫(yī)學(xué)在此方面為我們提供了新的思路和方法。胡黃連作為一種傳統(tǒng)保肝中藥，具有抗氧化、抗炎癥反應(yīng)、免疫調(diào)節(jié)等多種藥理學(xué)作用，胡黃連苷Ⅱ是從胡黃連的干根莖中提取的有效成分，分子量小，提純純度高，已經(jīng)發(fā)現(xiàn)其有抗炎癥作用。內(nèi)毒素誘導(dǎo)的葡萄膜炎(endotoxin induced uveitis，EIU)是用內(nèi)毒素中的脂多糖通過與脂多糖結(jié)合蛋白結(jié)合，提高血管通透性，通過激活補(bǔ)體系統(tǒng)和刺激血小板聚合因子釋放，從而引起組織炎癥。實(shí)驗(yàn)室常用的內(nèi)毒素為傷寒沙門氏菌或大腸桿菌內(nèi)毒素。實(shí)驗(yàn)動物一般為大鼠或小鼠，大鼠一般采用Lewis或SD鼠系。EIU模型是應(yīng)用最廣泛的葡萄膜炎動物模型，本模型是通過在實(shí)驗(yàn)動物腳掌皮下、腹腔或玻璃體腔注射細(xì)菌脂多糖，建立天然免疫機(jī)制誘導(dǎo)，釋放炎癥因子，也可有細(xì)胞介導(dǎo)的免疫反應(yīng)參與，EIU引起的葡萄膜炎持續(xù)時間短，表現(xiàn)為急性起病前葡萄膜炎。4小時出現(xiàn)炎癥，24小時達(dá)到高峰，之后炎癥反應(yīng)減輕�？赡軝C(jī)制為毒素的脂多糖進(jìn)入動物血液后形成脂多糖結(jié)合蛋白，然后再與Toll樣受體結(jié)合隨后傳遞到核轉(zhuǎn)錄因子NF—κB進(jìn)而啟動下游基岡的表達(dá)，最終引起組織炎癥反應(yīng)。我們即采用EIU葡萄膜炎模型來觀察胡黃連苷Ⅱ的抗炎作用。 方法:首先我們回顧分析了2012年03月至2013年09月于沈陽市第四人民醫(yī)院眼科就診并住院治療的葡萄膜炎患者195例235眼，患者均予以局部或全身糖皮質(zhì)激素治療，所有患者均隨訪6個月以上，觀察患者治療前后視力變化、炎癥反應(yīng)變化及局部全身出現(xiàn)的副作用。 隨后進(jìn)行基礎(chǔ)實(shí)驗(yàn)，研究胡黃連苷Ⅱ的抗炎作用。36只體重200~250g成年雄性SPF級SD大鼠隨機(jī)分成四組，每組9只，分別用20mg/kg和10mg/kg的胡黃連苷Ⅱ鼠尾靜脈注射兩次，其中3組第二次給藥后半小時予單側(cè)足底注射100μl濃度為100μg/100μl的LPS（Lipopolysaccharides），給藥后24小時前節(jié)裂隙燈照相，觀察炎癥變化并且評分，30gauge針收集（15-20μl）房水，細(xì)胞計數(shù)器計數(shù)，其余房水BCA試劑盒測房水蛋白濃度，取出眼球去核，OCT包埋冰凍切片，HE染色。 結(jié)果:病例回顧性分析中我們發(fā)現(xiàn)，195例葡萄膜炎患者的235眼中，男性70例80眼，女性125例155只眼，年齡9~78（平均37±23）歲，其中前葡萄膜炎109例（46.38%）、中間葡萄膜炎12例（5.12%）、后葡萄膜炎46例（19.57%）、全葡萄膜炎68例（28.93%）。治療后視力：眼前指數(shù)1眼（0.43%）、0.02~0.114眼（5.96%）、0.1~0.5101眼（42.98%）、0.5~0.8122眼（51.91%）、0.8~1.0及1.0以上7眼（2.98%）。血糖超過正常值41例（17.45%），停藥后血糖逐漸恢復(fù)正常；高血壓62例（26.38%），通過治療可以控制在正常血壓范圍內(nèi)；便潛血3例（1.28%），給與胃黏膜保護(hù)劑、糖皮質(zhì)激素減量停藥；癲癇發(fā)作1例（0.43%），經(jīng)治療后癥狀緩解；白細(xì)胞下降1例（0.43%），經(jīng)治療后痊愈；激素性青光眼3眼（1.28%），經(jīng)糖皮質(zhì)激素減量或停用局部激素眼藥水，并聯(lián)合降眼壓治療把眼壓控制在范圍內(nèi)；激素性白內(nèi)障9眼（3.83%），待葡萄膜炎穩(wěn)定后行白內(nèi)障超聲乳化術(shù)，術(shù)后視力恢復(fù)良好。 基礎(chǔ)實(shí)驗(yàn)中，細(xì)胞計數(shù)結(jié)果顯示20mg/kg組房水細(xì)胞少于LPS單純造模組（P0.05)）；10mg/kg組房水中細(xì)胞與LPS單純造模組比較無統(tǒng)計學(xué)差異（P0.05）。蛋白濃度測定結(jié)果20mg/kg組房水細(xì)胞少于LPS單純造模組（P0.05)）；10mg/kg組房水中蛋白濃度與LPS單純造模組比較無統(tǒng)計學(xué)差異（P0.05）。 結(jié)論:應(yīng)用激素治療各種非感染性葡萄膜炎具有一定效果，大多數(shù)患者病情好轉(zhuǎn)，但是糖皮質(zhì)激素治療期間均出現(xiàn)不同類型不同程度的副作用。胡黃連苷Ⅱ?qū)τ趦?nèi)毒素誘導(dǎo)的大鼠葡萄膜模型中具有抗炎作用。
[Abstract]:Objective: uveitis is one of the common ophthalmopathy and also one of the main blinding ophthalmopathy. Former uveitis is the most, followed by full uveitis. In developed countries, 5-20% patients are blinded by uveitis, and the proportion in developing countries may exceed 25%, because there are more opportunities for infectious diseases in the developing countries, such as arched. But at present, the main treatment for uveitis is the use of hormones, hormone treatment has a large side effect, and the ophthalmology may lead to hormone derived keratitis, glaucoma, and cataract. Systemic side effects include induced hypertension, diabetes, osteoporosis, gastrointestinal bleeding, seizures and so on, which all lead to the use of hormones. So we look for a drug that can effectively treat uveitis with small side effects. Chinese traditional medicine provides us with new ideas and methods in this field. Hu Huanglian, as a traditional Chinese medicine, has many pharmacological effects, such as antioxidation, anti-inflammatory reaction and immunoregulation. Lipopolysin II is an effective component extracted from rhizome of Rhizoma Coptidis, which has small molecular weight and high purity. It has been found that it has anti inflammatory effect. Endotoxin induced uveitis (EIU), which is induced by endotoxin, is a combination of lipopolysaccharide in endotoxin by binding with lipopolysaccharide binding protein to improve the permeability of blood vessels by activating a supplementary system. The release of platelet aggregation factors and stimulation of platelet aggregation causes inflammation of the tissues. The commonly used endotoxin in the laboratory is Salmonella typhi or Escherichia Coli Endotoxin. Experimental animals are generally rats or mice. Rats generally use the Lewis or SD mouse.EIU model to be the most widely used animal model of Vitis. This model is used in experimental animals. Subcutaneous, intraperitoneal or intravitreal injection of bacterial lipopolysaccharide to establish a natural immune mechanism, release inflammatory factors, and be involved in cell mediated immune responses. EIU induced uveitis has a short duration of uveitis, showing an acute onset of uveitis for.4 hours of inflammation, reaching a peak at 24 hours, and then alleviating the inflammatory response. The mechanism is that the lipopolysaccharide into the animal blood forms the lipopolysaccharide binding protein and then combines with the Toll like receptor and then transferred to the nuclear factor NF - kappa B and then initiates the expression of the downstream Keegan and eventually causes the inflammatory response of the tissue. We use the EIU uveitis model to observe the anti-inflammatory effect of the glucoside II.
Methods: first, we reviewed and analyzed 195 cases of uveitis in Fourth People's Hospital, Fourth People's Hospital of Shenyang from 03 months to 09 months in 2013. The patients were treated with local or systemic glucocorticoid. All patients were followed up for more than 6 months. The changes of visual acuity and the changes of inflammation were observed before and after treatment. Side effects of the local body.
Then the basic experiment was carried out to study the anti-inflammatory effect of hanthoside II,.36 only weight 200~250g adult male SPF SD rats were randomly divided into four groups, 9 rats in each group were injected two times with the tail vein of hanthoside 20mg/kg and 10mg/kg respectively, of which 3 groups were given second times after the injection of the sole plantar 100 L concentration of 100 u g/100 Mu L LPS ( Lipopolysaccharides), a slit lamp was taken 24 hours after the administration to observe the changes of inflammation and score. 30gauge needle was used to collect (15-20 L) water, cell counter count, and the rest aqueous BCA kit was used to measure the concentration of aqueous protein, remove the nucleus of the eyeball, OCT embedded frozen section, and HE staining.
Results: in the retrospective analysis, we found that 195 cases of uveitis were found in 235 eyes, male 70 cases, 80 eyes, 125 women in 155 eyes, age 9~78 (average 37 + 23) years, including 109 cases of anterior uveitis (46.38%), 12 cases of middle uveitis (5.12%), posterior Vitis 46 cases (19.57%), full uveitis. The index was 1 eyes (0.43%), 0.02~0.114 eye (5.96%), 0.1~0.5101 eye (42.98%), 0.5~0.8122 eye (51.91%), 0.8~1.0 and 7 eyes (2.98%). Blood sugar exceeded normal value in 41 cases (17.45%), blood sugar gradually returned to normal after withdrawal; hypertension 62 (26.38%) could be controlled within the normal blood pressure range; then the occult blood 3 (1.28%) was given to the gastric mucosa. 1 cases (0.43%) of epileptic seizures (0.43%) were relieved after treatment; the symptoms were relieved after treatment; 1 cases (0.43%) of leukocyte decreased (0.43%), 3 eyes (1.28%), glucocorticoid reduction or discontinuation of local hormone ophthalmic water, and intraocular pressure controlled by combined ophthalmic pressure reduction; 9 eyes of hormonal cataract (3) .83%) phacoemulsification is performed after uveitis is stable.
In the basic experiment, the cell count results showed that the water cells in the 20mg/kg group were less than the LPS model group (P0.05), and there was no statistical difference between the 10mg/kg group and the LPS simple model group (P0.05). The determination of protein concentration in the 20mg/kg group was less than that of the LPS simple model group (P0.05), and the protein concentration in the 10mg/kg group and LPS were simple. There was no statistical difference in the model (P0.05).
Conclusion: the use of hormones in the treatment of various non infectious uveitis has a certain effect, most patients have a better condition, but there are different types of side effects during the treatment of glucocorticoid.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R773.9

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 賀震旦,楊崇仁,王答祺,馮寶樹;西藏胡黃連的化學(xué)成分(英文)[J];云南植物研究;1993年01期

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