Zhang Ji, Song Yan, Wang Qiao-Han, Li Lin, Ji De, Gu Wei, Hao Min, Lu Tu-Lin, Mao Chun-Qin
School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China.
Jiangsu Key Laboratory of Chinese Medicine Processing, Nanjing University of Chinese Medicine, Nanjing 210023, China.
Zhongguo Zhong Yao Za Zhi. 2017 Jul;42(13):2538-2545. doi: 10.19540/j.cnki.cjcmm.20170610.001.
To compare the effects of Curcumae Rhizoma/vinegar-processed Curcumae Rhizomaon immune hepatic fibrosis, proliferation of HSC-T6, and expressions of α-SMA and Procollagen I. The immunological liver fibrosis model was prepared through intraperitoneal injection with porcine serum 0.5 mL in each rat, twice a week, for 14 weeks. Expressions of serum ALT, AST, PC-Ⅲ, IV-C, LN, HA and HYP, MDA in liver tissues were observed after administration of Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma (0.95, 1.90 g•kg⁻¹). The pathological changes in liver tissues were observed by HE staining. Masson staining and Sirius red staining were used to observe the expression of collagen in rat liver. HSC-T6 was cultured, and the proliferation of HSC-T6 was determined by MTT assay at different concentrations in 12, 24, 36, 48 h. The expressions of α-SMA and Procollagen I were detected by Real-time PCR. The results showed that expressions of serum ALT, AST, PC-Ⅲ, IV-C, LN and HA in Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma groups (0.95, 1.90 g•kg⁻¹) were significantly lower than model group; in terms of effect, vinegar-processed Curcumae Rhizoma group was superior to Curcumae Rhizoma group. Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma containing serum could inhibit the proliferation of HSC-T6 in a dose-effect and time-effect manner. Expressions of α-SMA and Procollagen I in HSC-T6 were decreased after 24 h, especially in 20% vinegar-processed Curcumae Rhizoma containing serum group (P<0.01). Both Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma could reduce immune hepatic fibrosis to varying extent. Their anti-hepatic fibrosis mechanism may be correlated with inhibition of the proliferation of HSC-T6, and reduction of the formation of extracellular matrix and promotion of its degradation.
比较姜黄/醋制姜黄对免疫性肝纤维化、肝星状细胞(HSC-T6)增殖及α-平滑肌肌动蛋白(α-SMA)和Ⅰ型胶原蛋白表达的影响。通过给每只大鼠腹腔注射0.5 mL猪血清制备免疫性肝纤维化模型,每周2次,共14周。给予姜黄/醋制姜黄(0.95、1.90 g•kg⁻¹)后,观察血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、Ⅲ型前胶原(PC-Ⅲ)、Ⅳ型胶原(IV-C)、层粘连蛋白(LN)、透明质酸(HA)和羟脯氨酸(HYP)的表达以及肝组织中丙二醛(MDA)的含量。通过苏木精-伊红(HE)染色观察肝组织的病理变化。采用Masson染色和天狼星红染色观察大鼠肝脏中胶原蛋白的表达。培养HSC-T6,通过MTT法在12、24、36、48 h测定不同浓度下HSC-T6的增殖情况。通过实时荧光定量聚合酶链反应(Real-time PCR)检测α-SMA和Ⅰ型胶原蛋白的表达。结果显示,姜黄/醋制姜黄组(0.95、1.90 g•kg⁻¹)血清中ALT、AST、PC-Ⅲ、IV-C、LN和HA的表达均显著低于模型组;在作用效果方面,醋制姜黄组优于姜黄组。含药血清的姜黄/醋制姜黄能以剂量效应和时间效应方式抑制HSC-T6的增殖。24 h后HSC-T6中α-SMA和Ⅰ型胶原蛋白的表达降低,尤其是含20%醋制姜黄血清组(P<0.01)。姜黄/醋制姜黄均能不同程度减轻免疫性肝纤维化。其抗肝纤维化机制可能与抑制HSC-T6增殖、减少细胞外基质形成并促进其降解有关。