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目的观察复元胶囊是否通过调节smad 3磷酸化水平促进骨关节炎软骨细胞合成Ⅱ型胶原(COL2A1)及蛋白多糖(ACAN)。方法体外培养人软骨瘤细胞株(SW1353),随机分为3组:空白组、白介素1β(IL-1β)组、IL-1β+复元胶囊组。以最适浓度15%复元胶囊含药血清进行实验干预。CCK8法观察细胞的生长情况;流式细胞术检测细胞周期;实时荧光定量PCR(q RT-PCR)检测细胞COL2A1、ACAN m RNA合成水平;Western blot检测细胞smad 3磷酸化水平。结果 IL-1β组的细胞生长曲线,细胞增殖率(25.50±0.63),COL2A1(0.523±0.009)、ACAN(0.201±0.035)m RNA及磷酸化smad 3(p-smad 3)(1.026±0.003)蛋白表达水平均低于空白组(P<0.05);IL-1β复元胶囊组的细胞生长曲线,细胞增殖率(37.55±0.71),COL2A1(0.701±0.068)、ACAN(0.901±0.230)m RNA及p-smad 3(1.527±0.003)蛋白表达水平均高于IL-1β组(P<0.01)。结论复元胶囊可以通过上调smad 3磷酸化水平,促进骨关节炎软骨细胞COL2A1及ACAN的合成,起到保护软骨细胞的作用。
Objective To observe whether Fuyuan capsule can promote the synthesis of collagen type Ⅱ (COL2A1) and proteoglycan (ACAN) by osteoarthritis chondrocytes by regulating the phosphorylation of smad 3. Methods Human chondrogenic cell line SW1353 was cultured in vitro and randomly divided into 3 groups: blank group, interleukin 1β (IL-1β) group and IL-1β + Fuyuan capsule group. The optimal concentration of 15% Fuyuan capsules containing serum for experimental intervention. CCK8 method was used to observe the cell growth. Flow cytometry was used to detect the cell cycle. Real-time quantitative PCR (q RT-PCR) was used to detect the levels of COL2A1 and ACAN m RNA. Western blot was used to detect the phosphorylation of smad 3. Results The cell growth curve, cell proliferation rate (25.50 ± 0.63), COL2A1 (0.523 ± 0.009), ACAN (0.201 ± 0.035) m RNA and p-smad 3 (1.026 ± 0.003) (P <0.05). The cell growth curve, cell proliferation rate (37.55 ± 0.71), COL2A1 (0.701 ± 0.068) and ACAN (0.901 ± 0.230) mRNA in IL- The protein level of p-smad 3 (1.527 ± 0.003) was higher than that of IL-1β group (P <0.01). Conclusion Fuyuan capsule can up-regulate the phosphorylation of smad 3 and promote the synthesis of osteoarthritis chondrocytes COL2A1 and ACAN and play a role in protecting chondrocytes.