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懸飲寧方對(duì)SPC
作者：徐振曄，劉建文，周美云【關(guān)鍵詞】 懸飲寧方
［摘要］ 目的：觀察中藥懸飲寧方抑制人肺腺癌細(xì)胞SPCA1浸潤(rùn)的作用及對(duì)S180腹水瘤小鼠腹膜病理形態(tài)學(xué)變化的影響。方法：在進(jìn)行小鼠腹膜間皮細(xì)胞分離培養(yǎng)的基礎(chǔ)上，將SPCA1人肺腺癌細(xì)胞調(diào)整加入復(fù)層培養(yǎng)，觀察形成的克隆數(shù)。取出S180腹水瘤小鼠腹膜，用透射電鏡觀察懸飲寧方各劑量組小鼠腹膜病理形態(tài)學(xué)的變化。結(jié)果：體外實(shí)驗(yàn)顯示加入含有懸飲寧生藥的培養(yǎng)液(50 μg/ml)后，SPCA1在瓊脂培養(yǎng)皿背面形成的癌細(xì)胞克隆數(shù)明顯減少。用懸飲寧治療S180腹水瘤小鼠后，取出腹膜，在電鏡下可觀察到，從低劑量開(kāi)始即可見(jiàn)小鼠腹膜間皮細(xì)胞排列整齊，隨著劑量的遞增這種現(xiàn)象更明顯，間皮細(xì)胞排列更加整齊，增厚；而生理鹽水組小鼠腹膜間皮細(xì)胞排列疏松，壞死。結(jié)論：中藥懸飲寧方有抑制SPCA1細(xì)胞浸潤(rùn)的作用，懸飲寧還可以改變S180腹水瘤小鼠排列疏松的腹膜間皮細(xì)胞，從而控制腹水瘤小鼠癌性積液生長(zhǎng)。
�。坳P(guān)鍵詞］ 懸飲寧方; SPCA1細(xì)胞; 間皮細(xì)胞; S180腹水瘤; 小鼠
Effects of Xuanyinning Recipe on invasion of SPCA1 cells and pathomorphological changes of peritoneum in mice inoculated with sarcoma 180
ABSTRACT Objective: To observe the effects of Xuanyinning Recipe (XYNR) in inhibiting SPCA1 cellular infiltration and on the pathomorphological changes of peritoneum in mice inoculated with sarcoma 180 (S180). Methods: On the bases of isolated culture of mouse peritoneal mesothelial cells, we adjusted and added the human lung adenocarcinoma cell line SPCA1 into the doublelayer culture medium to observe the number of clones formed. We also took out the peritoneum from the mice administered with three different dosages of XYNR and observed its pathomorphological changes with transmission electron microscope. Results: In the in vitro experiment, the number of clones of SPCA1 in culture medium with XYNR (50 μg/ml) decreased distinctly. In the in vivo experiment, it was observed that, in the peritoneum from the XYNRtreated mice inoculated with S180, the mesothelial cells arranged more and more regularly with the increasing of the dosage of XYNR, while the mesothelial cells in the peritoneum of the mice in the control group necrosed and arranged loosely. Conclusion: XYNR can inhibit the invasion of SPCA1 cells. It also can improve the loose arrangement of the peritoneal mesothelial cells in mice inoculated with S180, so as to inhibit the malignant effusion.
KEY WORDS Xuanyinning Recipe; SPCA1 cells; mesothelial cells; sarcoma 180; mice
研究資料表明，大約50%的癌癥轉(zhuǎn)移患者最終發(fā)生癌性積液［1］。而癌性積液的出現(xiàn)常常預(yù)示著疾病的進(jìn)展和生活質(zhì)量的下降，表明疾病進(jìn)入晚期，生存時(shí)間4～6個(gè)月［2］。我們?cè)谂R床及實(shí)驗(yàn)中觀察到中藥懸飲寧方能有效地控制癌性積液生長(zhǎng)［3］，為了進(jìn)一步探討懸飲寧方抑制癌性積液生長(zhǎng)的作用機(jī)制，我們進(jìn)行了懸飲寧方對(duì)SPCA1細(xì)胞浸潤(rùn)作用的體外實(shí)驗(yàn)，并通過(guò)電鏡觀察該方對(duì)S180腹水瘤小鼠腹膜組織病理形態(tài)的影響，現(xiàn)報(bào)告如下。
1 材料與方法
1.1 主要儀器與試劑 超凈工作臺(tái)，上海大華醫(yī)療儀器廠生產(chǎn)；CO2培養(yǎng)箱，日本平尺株式會(huì)社生產(chǎn)；倒置相差顯微鏡，日本Olympus公司制造；達(dá)氏修正依氏培養(yǎng)基（Dulbecco’s modified Eagle’s medium, DMEM）培養(yǎng)粉，由日本制藥株式會(huì)社生產(chǎn)；表皮生長(zhǎng)因子(epidermal growth factor,EGF)，由上海細(xì)胞生物學(xué)研究所提供；胰酶、酚紅、瓊脂粉、35 mm培養(yǎng)皿均由中國(guó)科學(xué)院實(shí)生細(xì)胞生物技術(shù)公司提供；順氯氨鉑，由中國(guó)齊魯制藥廠生產(chǎn)(批號(hào)：9386162)，規(guī)格20 mg／瓶；S180腹水瘤細(xì)胞株，引自中國(guó)科學(xué)院上海分院藥物研究所。
1.2 實(shí)驗(yàn)動(dòng)物 昆明種小鼠，雌雄各半，體質(zhì)量（20±2）g，6～8周齡，由中國(guó)科學(xué)院上海分院實(shí)驗(yàn)動(dòng)物中心提供（動(dòng)物合格證號(hào)：中科動(dòng)管005）。
1.3 實(shí)驗(yàn)藥物制備
1.3.1 體外實(shí)驗(yàn)用藥 懸飲寧液藥物按協(xié)定比例配方（由生白術(shù)、川椒目、葶藶子、貓人參等中藥組成），用三蒸水煎取，用乙醇明膠沉淀法制成，生藥終濃度為1 g／m1，pH值為6.8～7.2。
1.3.2 體內(nèi)實(shí)驗(yàn)用藥 懸飲寧藥液由上海中醫(yī)藥大學(xué)龍華醫(yī)院中藥藥房提供，每毫升含生藥濃度為2.8 g。用生理鹽水稀釋成2.49、1.2、0.6 g／ml。按《中藥藥理實(shí)驗(yàn)方法學(xué)》附表“人和動(dòng)物間接體表面積折算表［4］”，計(jì)算出高、中、低不同劑量組的用藥劑量分別為52.36、32.36、20 g／kg；每只小鼠每日用藥量分別為1.047、0.647、0.4 g，灌胃用藥。順氯氨鉑，用生理鹽水稀釋成0.
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