Chinese Journal of Tissue Engineering Research ›› 2023, Vol. 27 ›› Issue (23): 3682-3692.doi: 10.12307/2023.570
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Li Gang1, Song Yun2, Liang Guoguang3
Received:
2022-07-26
Accepted:
2022-10-18
Online:
2023-08-18
Published:
2023-01-14
About author:
Li Gang, MD, Lecturer, Associate chief physician, Guangdong Vocational College of Food and Drug, Guangzhou 510000, Guangdong Province, China
Supported by:
CLC Number:
Li Gang, Song Yun, Liang Guoguang. Wenyang Tongluo capsule intervenes with inflammation and angiogenesis in rat osteoarthritic chondrocytes[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(23): 3682-3692.
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2.1.3 各组大鼠膝关节软骨大体观察 正常组大鼠行动机警,肢体协调性良好,大鼠未见明显的行为异常;模型组大鼠的肢体灵活性明显下降,出现跛行的状况,活动量明显减少,大鼠的病灶膝关节部位严重肿胀,大鼠舐足次数明显增多;与模型组相比,药物组大鼠的临床症状明显改善,跛行状况明显缓解,舐足次数明显减少;与药物组相比,抑制剂组大鼠病灶部位明显肿胀,大鼠舐足次数明显增多;与抑制剂组相比,干扰组大鼠的膝关节的肿胀以及舐足行为有所改善。 将大鼠处死后,大体观察膝关节软骨,结果显示:正常组大鼠膝关节表面光滑,色泽透明,干燥,无增生;模型组大鼠关节软骨色泽暗淡,表面粗糙,部分区域出现米黄色病变,软骨层缺损明显,出现部分糜烂状况;与模型组相比,药物组、抑制剂组、干扰组大鼠关节性状明显好转,以温阳通络胶囊组改善最为明显;与抑制剂组相比,干扰组大鼠膝关节状况有了较为明显改善,见图2。"
2.1.8 各组大鼠膝关节软骨组织中miR-140、VEGF、Bcl-2、Bax的表达 qRT-PCR检测:与正常组相比,模型组、药物组、抑制剂组、干扰组VEGF、Bax的mRNA表达明显升高,miR-140、Bcl-2的mRNA表达明显降低(P < 0.05);与模型组相比,药物组、抑制剂组、干扰组VEGF、Bax的mRNA表达明显降低(P < 0.05),miR-140、Bcl-2的mRNA表达明显升高(P < 0.05);与药物组相比,抑制剂组、干扰组VEGF、Bax的mRNA表达明显升高(P < 0.05),miR-140、Bcl-2的mRNA表达明显降低(P < 0.05);与抑制剂组相比,干扰组VEGF、Bax的mRNA表达明显降低(P < 0.05),miR-140、Bcl-2的mRNA表达明显升高(P < 0.05),见表2。 "
Western blot检测:与正常组相比,模型组、药物组、抑制剂组、干扰组VEGF、Bax的蛋白表达明显升高(P < 0.05),Bcl-2的蛋白表达明显降低(P < 0.05);与模型组相比,药物组、抑制剂组、干扰组VEGF、Bax的蛋白表达明显降低(P < 0.05),Bcl-2的蛋白表达明显升高(P < 0.05);与药物组相比,抑制剂组、干扰组VEGF、Bax的蛋白表达明显升高(P < 0.05),Bcl-2的蛋白表达明显降低(P < 0.05);与抑制剂组相比,干扰组VEGF、Bax的蛋白表达明显降低(P < 0.05),Bcl-2的蛋白表达明显升高(P < 0.05),见图9。"
2.2.7 各组软骨细胞中miR-140、VEGF、Bcl-2、Bax的表达 qRT-PCR检测:与正常组相比,模型组、药物组、抑制剂组、干扰组VEGF、Bax的mRNA表达明显升高(P < 0.05),miR-140、Bcl-2的mRNA表达明显降低(P < 0.05);与模型组相比,药物组、抑制剂组、干扰组VEGF、Bax的mRNA表达明显降低(P < 0.05),miR-140、Bcl-2的mRNA表达明显升高(P < 0.05);与药物组相比,抑制剂组、干扰组VEGF、Bax的mRNA表达明显升高(P < 0.05),miR-140、Bcl-2的mRNA表达明显降低(P < 0.05);与抑制剂组相比,干扰组VEGF、Bax的mRNA表达明显降低(P < 0.05),miR-140、Bcl-2的mRNA表达明显升高(P < 0.05),见表3。"
Western blot检测:与正常组相比,模型组、药物组、抑制剂组、干扰组VEGF、Bax的蛋白表达明显升高(P < 0.05),Bcl-2的蛋白表达明显降低(P < 0.05);与模型组相比,药物组、抑制剂组、干扰组VEGF、Bax的蛋白表达明显降低(P < 0.05),Bcl-2的蛋白表达明显升高(P < 0.05);与药物组相比,抑制剂组、干扰组VEGF、Bax的蛋白表达明显升高(P < 0.05),Bcl-2的蛋白表达明显降低(P < 0.05);与抑制剂组相比,干扰组VEGF、Bax的蛋白表达明显降低(P < 0.05),Bcl-2的蛋白表达明显升高(P < 0.05),见图16,表4。"
2.2.8 温阳通络胶囊通过miR-140/VEGF轴对大鼠骨关节炎软骨细胞的影响机制 在体内,当关节软骨组织处于关节炎病理状态中时,软骨组织中的miR-140的表达明显下降,VEGF的含量明显升高,白细胞介素6和肿瘤坏死因子α的分泌升高,抗凋亡蛋白Bcl-2与促凋亡蛋白Bax的表达失衡,大鼠软骨组织的解剖学损伤、病理损伤明显加重,软骨组织的细胞凋亡随之升高,软骨中的新生血管数量明显增多,促进关节炎的病程进展;当以温阳通络胶囊进行干预时,可明显逆转这些参数,达到改善骨关节炎的治疗效果。 在体外,软骨细胞ATDC5受白细胞介素1β炎性递质刺激,细胞中miR-140与VEGF的表达失衡,miR-140的表达明显下降,VEGF的含量明显升高,细胞的增殖活性明显下降,凋亡率明显升高,细胞上清液中白细胞介素6和肿瘤坏死因子α的含量升高,人脐静脉内皮细胞成管性能明显升高,而当以温阳通络胶囊进行干预时可明显逆转这些参数,明显改善软骨细胞ATDC5的增殖活性、抑制其凋亡和促血管生成的能力,达到干预关节炎的目的。"
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