目的 研究豬苓多糖抑制胃黏膜上皮細胞（GES-1）間充質(zhì)轉(zhuǎn)化的作用和機制。方法 應(yīng)用N-甲基-N'-硝基-N-亞硝基（MNNG，40 μmol·L^-1）誘導(dǎo)GES-1細胞間充質(zhì)轉(zhuǎn)化模型，造模同時給予豬苓多糖（1.25、2.50、5.00、10.00 mg·mL^-1）處理24、48 h后，CCK-8法檢測細胞增殖能力；造模同時給予豬苓多糖（5.00 mg·mL^-1）處理48 h后，實時定量PCR（qRT-PCR）法檢測豬苓多糖對N-cadherin、Snail、Twist、Fibronection及Vmention mRNA表達的影響；Western blotting法檢測E-cadherin、N-cadherin、Snail、Twist、Fibronetin、Vimentin、STAT3、p-STAT3、JAK2、p-JAK2蛋白表達。裸鼠分別sc經(jīng)MNNG 40 μmol·L^-1處理48 h后的GES-1細胞（5×10⁷·mL^-1 ，0.2 mL）、胃癌細胞SGC7901（5×10⁷·mL^-1 ，0.2 mL ，陽性對照），1個月后觀察各組成瘤及體質(zhì)量情況，注射局部進行HE染色后行組織病理學(xué)觀察，驗證GES-1細胞經(jīng)MNNG處理后是否達到腫瘤階段。結(jié)果 與模型組比較，隨豬苓多糖濃度的增加，GES-1細胞增殖能力逐漸上升，到達5 mg·mL^-1時增殖能力最高，差異顯著（P<0.01、0.001）；豬苓多糖組N-cadherin、Snail、Twist、Fironetion及Vimentin的mRNA表達均顯著下降（P<0.05、0.01、0.001）；豬苓多糖組E-cadherin蛋白表達顯著上升（P<0.05），N-cadherin、Snail、Twist、Fibronetion、Vimentin及通路蛋白STAT3、p-STAT3、JAK2、p-JAK2表達顯著下降（P<0.05、0.01、0.001）。對照組和MNNG處理的GES-1組未見瘤體形成，陽性對照SGC7901組注射后1周左右局部出現(xiàn)瘤體，1個月瘤體長至6 cm左右；MNNG處理的GES-1組裸鼠精神較差，體質(zhì)量明顯減輕，SGC7901組裸鼠狀態(tài)差，體質(zhì)量大幅減輕；對照組和MNNG組病理染色顯示為正常的皮膚組織，陽性對照組顯示為腫瘤組織。結(jié)論 豬苓多糖可能通過抑制JAK2-STAT3通路干預(yù)MNNG誘導(dǎo)的GES-1細胞上皮間充質(zhì)化。

Objective To study the inhibitory effect of polyporus polysaccharides (PPS) on mesenchymal transformation of gastric epithelial cells (GES-1) and its preliminary mechanism. Methods GES-1 cell mesenchymal transition model was induced by N-methyl-N'-nitro-N-nitroso (MNNG, 40 μmol·L^-1). The model was established and treated with PPS (1.25, 2.50, 5.00, 10.00 mg·mL^-1) for 24 and 48 h. The cell proliferation ability was measured using the CCK8 method.After 48 hours of treatment with PPS(5.00 mg·mL^-1), the effects of PPS on N-cadherin, Snail, Twist, Fibronection, and Vmention mRNA expression were detected using real-time quantitative PCR (qRT-PCR), and Western blotting was used to detect the expression of E-cadherin, N-cadherin, Snail, Twist, Fibronetin, Vimentin, STAT3, p-STAT3, JAK2, and p-JAK2 proteins. Nude mice were subjected to GES-1 cells (5×10⁷·mL^-1, 0.2 mL) treated with MNNG 40 μmol·L^-1 for 48 hours and gastric cancer cells SGC7901 (5×10⁷·mL^-1, 0.2 mL, positive control), after one month, observed the tumor and body mass of each component, and observed the histopathological changes after HE staining at the injection site to verify whether GES-1 cells had reached the tumor stage after MNNG treatment. Results Compared with the model group, as the concentration of PPS increased, the proliferation ability of GES-1 cells gradually increased, reaching the highest proliferation ability at 5 mg·mL^-1, with significant differences (P< 0.01, 0.001); The mRNA expression of N-cadherin, Snail, Twist, Fironeton, and Vimentin in the PPS group was significantly reduced (P< 0.05, 0.01, 0.001); The expression of E-cadherin protein was significantly increased in the polyporus polysaccharides group (P< 0.05), while the expression of ^N-cadherin, ^Snail, ^Twist, ^Fibronection, ^Vimentin, and pathway proteins STAT3, p-STAT3, JAK2, and p-JAK2 were significantly decreased (P< 0.05, 0.01, 0.001). The control group and MNNG treated GES-1 group showed no tumor formation, while the positive control SGC7901 group showed local tumor formation around one week after injection, and the tumor grew to about 6 cm in length after one month; The GES-1 group of nude mice treated with MNNG showed poor mental state and significantly reduced body mass, while the SGC7901 group showed poor state and significantly reduced body weight. The pathological staining of the control group and MNNG group showed normal skin tissue, while the positive control group showed tumor tissue. Conclusion PPS may interfere with MNNGinduced epithelial mesenchymal transition of GES-1 cells by inhibiting JAK2-STAT3 pathway.

R285.5

國家自然科學(xué)基金青年項目（81801625）