Effect of Duhuo Jisheng Decoction on PERK/Bip signaling pathway in cartilage of a rat model of knee osteoarthritis

doi:10.3969/j.issn.2095-4344.0391

Abstract

Abstract:

BACKGROUND: Preliminary study has shown that Duhuo Jisheng Decoction (DHJSD) is an effective prescription for osteoarthritis. But the underling mechanism remains unclear.
OBJECTIVE: To observe the effect of DHJSD on the key regulating factors PERK, Bip, eIF-2α, ATF-4, GADD153, Caspase-9 and Caspase-3 in PERK/Bip signaling pathway in cartilage of rats with knee osteoarthritis, so as to explore its treatment mechanism.
METHODS: After 1 week of acclimation, 80 Sprague-Dawley rats were randomly divided into normal group (n=20) and model rats (n=60). Osteoarthritis was induced by injecting 0.2 mL of 4% papain solution in both knees. The rat models were randomly divided into three groups, including model, treatment and positive control groups (n=20 per group). The normal and model groups received treatment of normal saline. The treatment and positive control groups were given DHJSD (9.3 g/(kg•d)) and glucosamine hydrochloride capsule 0.15 g/(kg•d) via gavage, respectively, for four courses (two weeks a course). After every two courses, the animals were sacrificed and the right tibial plateau was obtained. The mRNA and protein levels of PERK, Bip, eIF-2α, ATF-4, GADD153, Caspase-9 and Caspase-3 were measured by RT-PCR and immunohistochemistry, respectively.
RESULTS AND CONCLUSION: RT-PCR results showed that DHJSD and glucosamine hydrochloride capsule could time-dependently inhibit the mRNA expression of PERK, Bip, eIF-2α, ATF-4, GADD153, Caspase-9 and Caspase-3 (P < 0.05 or P < 0.01). There was no significant difference between treatment and positive control groups (P > 0.05). Immunohistochemistry results found that DHJSD and glucosamine hydrochloride capsule could time-dependently inhibit the protein expression of PERK, Bip, eIF-2α, ATF-4, GADD153, Caspase-9 and Caspase-3 (P < 0.05 or P < 0.01). There was no significant difference between treatment and positive control groups (P > 0.05). These results suggest that DHJSD can inhibit the apoptosis of chondrocytes caused by endoplasmic reticulum stress through regulating the PERK/Bip signaling pathway.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Heracleum, Osteoarthritis, Apoptosis, Tissue Engineering

CLC Number:

R318

Chen Jun1, Wu Guang-wen2, Xu Hui-feng2, Zheng Chun-song3, Li Xi-hai3, Qiu Jian-qing3, Liu Shu-ru3, . Effect of Duhuo Jisheng Decoction on PERK/Bip signaling pathway in cartilage of a rat model of knee osteoarthritis[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(28): 4493-4500.

Figures/Tables 4

2.1 实验动物数量分析在干预过程中，模型组麻醉意外致死1只，治疗组在灌胃过程中出现腹泻致死1只，均给予补全后，所有动物进入结果分析。 2.2 独活寄生汤对大鼠关节软骨组织PERK、Bip、eIF-2α、ATF-4、GADD153、Caspase-9和Caspase-3 mRNA表达的影响如图1 A-H，4周时，与正常组相比，模型组4周中PERK、Bip、eIF-2α、ATF-4、GADD153、Caspase-9和Caspase-3 mRNA的表达量均升高，差异有显著性意义(P < 0.01)；治疗组和阳性对照组PERK、Bip、eIF-2α、ATF-4、GADD153、Caspase-9和Caspase-3 mRNA的表达量均逐渐降低，与模型组相比，差异均有显著性意义(P < 0.05)，但治疗组与阳性对照组之间无显著性差异(P > 0.05)。随着治疗时间的延长，8周时，与正常组相比，模型组中PERK、Bip、eIF-2α、ATF-4、GADD153、Caspase-9和Caspase-3 mRNA的表达量均明显升高，差异有显著性意义(P < 0.01)；治疗组和阳性对照组中PERK、Bip、eIF-2α、ATF-4、GADD153、Caspase-9和Caspase-3 mRNA表达量均进一步降低，与模型组8周相比，差异有显著性意义(P < 0.01)，分别与治疗组4周和阳性对照组4周相比，差异具有显著性意义(P < 0.05)，但治疗组与阳性对照组之间无显著性意义(P > 0.05)。 2.3 独活寄生汤对大鼠关节软骨组织PERK、Bip、eIF-2α、ATF-4、GADD153、Caspase-9和Caspase-3蛋白表达的影响 PERK、Bip、eIF-2α、ATF-4、GADD153、Caspase-9和Caspase-3阳性染色呈棕黄色或棕褐色，主要位于细胞胞浆中(见图2 A-H、图3 A-H、图4 A-H、图5 A-H、图6 A-H、图7 A-H、图8 A-H)。比较各组阳性率(图2I、图3I、图4I、图5I、图6I、图7I、图8I)可见各组对PERK、Bip、eIF-2α、ATF-4、GADD153、Caspase-9和Caspase-3蛋白表达的影响与其对PERK、Bip、eIF-2α、ATF-4、GADD153、Caspase-9和Caspase-3 mRNA的影响具有相同的趋势，与正常组相比，模型组中PERK、Bip、eIF-2α、ATF-4、GADD153、Caspase-9和Caspase-3阳性率均升高，差异均有显著性意义(P < 0.01)；治疗组和阳性对照组"

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