肺纖方顆粒對(duì)鹽酸平陽(yáng)霉素致肺間質(zhì)纖維化大鼠血清轉(zhuǎn)化生長(zhǎng)因子β

1、肺纖方顆粒對(duì)鹽酸平陽(yáng)霉素致肺間質(zhì)纖維化大鼠血清轉(zhuǎn)化生長(zhǎng)因子         作者：張偉, 姜良鐸, 張曉梅, 吳建軍, 鄧媛, 魯香鳳 【摘要】 觀察肺纖方顆?？勾笫蠓伍g質(zhì)纖維化的作用，及其對(duì)大鼠血清轉(zhuǎn)化生長(zhǎng)因子1（transforming growth factor1, TGF1）和血小板衍生生長(zhǎng)因子（plateletderived growth factor，PDGF）的影響。方法：72          

2、60;                作者：張偉, 姜良鐸, 張曉梅, 吳建軍, 鄧媛, 魯香鳳【摘要】  觀察肺纖方顆粒抗大鼠肺間質(zhì)纖維化的作用，及其對(duì)大鼠血清轉(zhuǎn)化生長(zhǎng)因子1（transforming growth factor1, TGF1）和血小板衍生生長(zhǎng)因子（plateletderived growth factor，PDGF）的影響。方法：72只雄性Wistar大鼠經(jīng)氣管插管灌注鹽酸平陽(yáng)霉素1 mg/kg復(fù)制大鼠肺間質(zhì)纖維化模

3、型，隨機(jī)分入模型組、潑尼松組和肺纖方組，每組24只；15只雄性Wistar大鼠經(jīng)氣管插管灌注等體積生理鹽水作為假手術(shù)組。造模后24 h開始常規(guī)灌胃給藥，潑尼松組給予醋酸潑尼松5 mg/kg，肺纖方組給予肺纖方顆粒1.25 g/kg，模型組和假手術(shù)組給予等體積生理鹽水，1次/d。分別于第14、28、45 天取材，假手術(shù)組每批5只，其余各組每批8只。檢測(cè)其肺組織病理改變及血清中TGF1和PDGF的水平。結(jié)果：與模型組和潑尼松組比較，在第45天時(shí)病理檢測(cè)顯示肺纖方組肺間質(zhì)纖維化的程度較輕；血清TGF1和PDGF檢測(cè)結(jié)果顯示，模型組大鼠兩種因子水平升高，第45天時(shí)升高最明顯，并高于假手術(shù)組（P<

4、0.05）。肺纖方組與潑尼松組的TGF1變化與模型組相似（P>0.05），兩組間TGF1變化比較差異無(wú)統(tǒng)計(jì)學(xué)意義（P>0.05）。肺纖方組PDGF水平在第14天時(shí)與其余各組比較顯著升高（P<0.01），然后持續(xù)下降，至第45天與假手術(shù)組接近（P=0.792）；而模型組和潑尼松組PDGF水平皆隨時(shí)間延長(zhǎng)逐漸上升，第45天時(shí)均顯著高于假手術(shù)組（P<0.01）。結(jié)論：肺纖方有一定的減輕鹽酸平陽(yáng)霉素致大鼠肺間質(zhì)纖維化的作用，其作用機(jī)制可能與對(duì)血清PDGF水平的調(diào)節(jié)作用有關(guān)。 【關(guān)鍵詞】  肺纖方顆粒; 肺疾病, 間質(zhì)性; 肺纖維化; 轉(zhuǎn)化生長(zhǎng)因子1; 血小板衍生生長(zhǎng)因

5、子; 大鼠Objective: To observe the effects of Feixian Formula, a compound traditional Chinese herbal medicine for treating pulmonary fibrosis, on bleomycininduced pulmonary fibrosis in rats, and its influence on serum transforming growth factor1 (TGF1) and plateletderived growth factor (PDGF).Methods: S

6、eventytwo male Wistar rats were infused with bleomycin (1 mg/kg) through tracheal intubation to induce pulmonary fibrosis, and they were randomly divided into untreated group (n=24), prednisonetreated group (n=24) and Feixian Formulatreated group (n=24). Fifteen male Wistar rats of the shamoperated

7、group were infused with equivalent normal saline. Twentyfour hours after operation, prednisone (5 mg/kg) and Feixian Formula (1.25 g/kg) were given to the prednisonetreated group and Feixian Formulatreated group respectively by intragastric administration once a day. Equivalent saline was administer

8、ed to rats of the untreated group and shamoperated group. On the 14th, 28th and 45th day, 5 rats in the shamoperated group and 8 rats in each of the other three groups were dissected to observe pathologic changes of the lung tissues, and the levels of serum TGF1 and PDGF were determined by enzymelin

9、ked immunosorbent assay.Results: At the 45th day, the degree of pulmonary interstitial fibrosis was lesser in rats of the Feixian Formulatreated group as compared with those of the untreated group and prednisonetreated group. The levels of serum TGF1 and PDGF were increased, and were significantly h

10、igher than those of the shamoperated group, especially on the 45th day (P<0.05). Changes of TGF1 level in the prednisonetreated group and the Feixian Formulatreated group were similar to untreated group (P>0.05), and there was no significant difference between the prednisonetreated group and t

11、he Feixian Formulatreated group (P>0.05). PDGF in the Feixian Formulatreated group reached the highest level on the 14th day, significantly higher than those of the other three groups (P<0.01). Then it decreased, and was close to that of the shamoperated group on the 45th day (P=0.792). The le

12、vels of PDGF in untreated group and prednisonetreated group were increased depending on time, and were obviously higher than that of the shamoperated group on the 45th day (P<0.01).Conclusion: Feixian Formula can relieve bleomycininduced pulmonary fibrosis in rats, and the mechanism of its action

13、 may be related to downregulating serum PDGF.Keywords: Feixian Formula; lung diseases, interstitial; pulmonary fibrosis; transforming growth factor 1；plateletderived growth factor; rats    肺間質(zhì)纖維化是許多不同病因所致肺間質(zhì)疾病的共同結(jié)局，其發(fā)病機(jī)制尚不完全清楚，至今沒有理想的治療方法。本研究使用鹽酸平陽(yáng)霉素（bleomycin A5, BLM）1 mg/kg經(jīng)氣管插管快速灌注

14、的方法復(fù)制大鼠肺間質(zhì)纖維化模型，研究中藥制劑肺纖方顆粒對(duì)BLM致肺間質(zhì)纖維化大鼠肺組織病理、血清轉(zhuǎn)化生長(zhǎng)因子1（transforming growth factor1, TGF1）和血小板衍生生長(zhǎng)因子（plateletderived growth factor, PDGF）水平的影響。1  材料與方法1.1  實(shí)驗(yàn)材料清潔級(jí)健康雄性Wistar大鼠87只，體質(zhì)量180200 g，由北京維通利華公司提供，許可證號(hào)為京動(dòng)許字（1999）第039號(hào)。注射用鹽酸平陽(yáng)霉素，8 mg/支，天津太和制藥有限公司生產(chǎn)，批號(hào)為050402。肺纖方顆粒劑的制作由北京師范大學(xué)教育部資源藥物工程研

15、究中心承擔(dān)，制備工藝符合新藥申報(bào)要求，藥品規(guī)格為每劑生藥89.5 g，制成25.14 g顆粒，每克顆粒相當(dāng)于生藥3.56 g。醋酸潑尼松片，5 mg/片，天津力生制藥股份有限公司生產(chǎn)，批號(hào)為0503019。18號(hào)套管針、光源、紗布、35度傾斜鼠板、LEICA RM2135石蠟切片機(jī)（德國(guó)）、LEICA DMLB顯微鏡（德國(guó)）、OLYMPUS全自動(dòng)顯微攝像系統(tǒng)（日本）、Specotrat酶標(biāo)比色儀（SLT公司）。1.2  實(shí)驗(yàn)方法87只雄性Wistar大鼠經(jīng)腹腔注射10水合氯醛(3.5 ml/kg體質(zhì)量)麻醉后仰面固定于35度傾斜鼠板上，將光源直接照射于大鼠咽喉部皮膚表面，左手墊紗布向

16、外向上提起舌頭，可見咽喉部有一明顯的亮點(diǎn)，從大鼠口腔，右手水平插入18號(hào)套管針到達(dá)咽喉部時(shí)，壓低針尾，使針頭抬高1530度，輕輕插入氣管，長(zhǎng)度約為針長(zhǎng)的3/4，拔出針芯。將棉球薄片置于套管針外口，若棉球薄片隨呼吸前后擺動(dòng)，則說明套管針已經(jīng)插入氣管，若棉球薄片不隨呼吸前后擺動(dòng)，可能套管針誤入食道，應(yīng)立即拔出重新插入。其中72只大鼠按1 mg/kg體質(zhì)量劑量1向氣管插管內(nèi)快速注射2 mg/ml BLM生理鹽水溶液，15只假手術(shù)組動(dòng)物注射等體積的生理鹽水，注射后繼續(xù)向氣管內(nèi)快速推注0.30.5 ml空氣，并立即將大鼠直立旋轉(zhuǎn)，以使藥液在雙肺內(nèi)均勻分布。動(dòng)物清醒后隨意進(jìn)食，將BLM造模組大鼠隨機(jī)分為模型組、潑尼松組和肺纖方組，每組各24只。造模24 h后開始灌胃給藥，1次/d，潑尼松組給予醋酸潑尼松5 mg/kg（藥液濃度為0.5 mg/ml），肺纖方組給予肺纖方顆粒1.25 g/kg（相當(dāng)于生藥4.475 g/kg，藥液濃度為0.125 g/ml），模型組和假手術(shù)組給予等量生理鹽水。造模后第14天、第28天和第45天分3批取材，假手術(shù)組每批5只，其余各組