Yan Shuai, Yang Lan, Yue Yin-Zi, Li Wen-Lin, Zeng Li, Yue Jin, Mao Chun-Qin
Suzhou Hospital of Traditional Chinese Medicine, Suzhou, Jiangsu Province, 215009, China.
First Clinical Medical College of Nanjing University of Chinese Medicine, Nanjing, 210023, China.
Chin J Integr Med. 2016 Aug;22(8):629-34. doi: 10.1007/s11655-015-2180-8. Epub 2015 Sep 30.
To study the effect of ligustrazine nanoparticles nano spray (LNNS) on transforming growth factor β (TGF-β)/Smad signal protein of rat peritoneal mesothelial cells (RPMC) induced by tumor necrosis factor α (TNF-α), and the anti-adhesion mechanism of LNNS in the abdominal cavity.
The primary culture and subculture of rat peritoneal mesothelial cells (RPMC) was processed by trypsin digestion method in vitro. The third generation was identifified for experiment and divided into 5 groups: a blank group: RPMC without treatment; a control group: RPMC stimulated with TNF-α; RPMC treated by a low-dosage LNNS group (2.5 mg/L); RPMC treated by a medium-dosage LNNS group (5 mg/L); and RPMC treated by a high-dosage LNNS group (10 mg/L). Reverse transcription-polymerase chain reaction was applied to test the expression of fifibronectin, collagen I (COL-I), TGF-β mRNA, and Western blot method to test the Smad protein 7 expression of RPMC.
Compared with the blank group, a signifificant elevation in fifibronectin (FN), COL-I and TGF-β mRNA expression of RPMC were observed in the control group (P<0.05). Compared with the control group, LNNS suppressed the expressions of FN, COL-I and TGF-β mRNA in a concentrationdependent manner (P<0.05). The expression of Smad7 protein of RPMC was down-regulated by TNF-α stimulation, and up-regulated with the increase of LNNS dose (P<0.05).
TNF-α may induce changes in RPMC's viability, leading to peritoneal injury. LNNS could reverse the induction of fifibrosis related cytokine FN, COL-I and TGF-β, up-regulating the expression of Smad7 by TNF-α in RPMC, thus attenuate peritoneal injury by repairing mesothelial cells.
研究川芎嗪纳米颗粒纳米喷雾剂(LNNS)对肿瘤坏死因子α(TNF-α)诱导的大鼠腹膜间皮细胞(RPMC)转化生长因子β(TGF-β)/Smad信号蛋白的影响,以及LNNS在腹腔内的抗粘连机制。
采用胰蛋白酶消化法体外进行大鼠腹膜间皮细胞(RPMC)的原代培养和传代培养。取第三代细胞进行实验鉴定,并分为5组:空白组:未处理的RPMC;对照组:用TNF-α刺激的RPMC;低剂量LNNS处理组(2.5 mg/L)的RPMC;中剂量LNNS处理组(5 mg/L)的RPMC;高剂量LNNS处理组(10 mg/L)的RPMC。应用逆转录-聚合酶链反应检测纤连蛋白、Ⅰ型胶原(COL-I)、TGF-β mRNA的表达,采用蛋白质印迹法检测RPMC中Smad蛋白7的表达。
与空白组相比,对照组RPMC的纤连蛋白(FN)、COL-I和TGF-β mRNA表达显著升高(P<0.05)。与对照组相比,LNNS以浓度依赖性方式抑制FN、COL-I和TGF-β mRNA的表达(P<0.05)。TNF-α刺激使RPMC中Smad7蛋白表达下调,随LNNS剂量增加而上调(P<0.05)。
TNF-α可能诱导RPMC活力改变,导致腹膜损伤。LNNS可逆转纤维化相关细胞因子FN、COL-I和TGF-β的诱导,上调TNF-α在RPMC中对Smad7的表达,从而通过修复间皮细胞减轻腹膜损伤。