Study on Protective Effects of Longbie Capsule Contained Serum on the Apoptosis of Chondrocytes Induced by YAP Inhibitor
- VernacularTitle:龙鳖胶囊含药血清对YAP抑制剂诱导人软骨细胞凋亡的保护作用及机制研究
- Author:
Guihong LIANG
1
,
2
;
Hetao HUANG
1
;
Jianke PAN
1
;
Lingfeng ZENG
1
,
2
;
Weiyi YANG
1
;
Minghui LUO
1
;
Yuan YANG
1
,
2
;
Hongyun CHEN
1
;
Yanhong HAN
1
;
Jinlong ZHAO
1
;
Jun LIU
1
Author Information
1. Dept. of Orthopaedics，the Second Affiliated Hospital of Guangzhou University of TCM （Guangdong Provincial Hospital of TCM ），Guangzhou 510120，China
2. Research Team of Bone and Joint Degeneration and Injury，Guangdong Provincial Academy of Chinese Medicina l Sciences，Guangzhou 510120，China
- Publication Type:Journal Article
- Keywords:
Longbie capsule;
Contained serum;
Osteoarthritis;
Chondrocytes;
YAP inhibitor;
Apoptosis;
Autophagy
- From:
China Pharmacy
2021;32(12):1442-1448
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE：To ex plore the protective effects of Longbie capsule contained serum （called“LBJN”for short ）on the apoptosis of chondrocytes induced by YAP inhibitor verteporfin and its mechanism. METHODS ：Primary human knee osteoarthritis（OA）chondrocytes were extracted by two-step enzymatic digestion ，and then identif ied by toluidine blue staining and type Ⅱ collagen immunofluorescence staining. The effects of 2，5 μmol/L verteporfin alone or combined with 5％LBJN on cell apoptosis were detected by flow cytometry. Solvent control （0.1％ DMSO）and 5％ LBJN were set. Western blot assay was adopted to detect the expression of apoptosis related proteins （YAP，Bcl-2，cleaved-caspase-3） after treated with 0.1％DMSO（solvent control ），2 μmol/L verteporfin，2 μmol/L verteporfin+5％LBJN 和 0（blank control ），2.5％ LBJN and 5％ LBJN for 48 h. The expression of autophagy related proteins （mTOR，Beclin-1，LC3A/B） after treated with 0 （blank control ），2.5％，5％ LBJN for 48 h were det ected by Western blot assay. RESULTS ：The isolated cells accorded with the characteristics of chondrocytes. Compared with 0.1％DMSO， the apoptosis rates of cells were increased significantly after treated with 2，5 μmol/L verteporfin（P＜0.05），and the effects of the two concentrations were similar （P＞0.05）. Compared with verteporfin alone ，2，5 μmol/L verteporfin combined with 5％LBJN could significantly decrease the apoptotic rate of cells （P＜0.05）. Compared with 0.1％DMSO，the protein expression of YAP and Bcl-2 were decreased significantly after treated with 2 μ mol/L verteporfin （P＜0.05）， while the protein expression of cleaved-caspase-3 were increased significantly （P＜0.05）. Compared with 2 μmol/L verteporfin，protein expression of YAP and Bcl-2 were increased significantly after treated with 2 μmol/L verteporfin+5％LBJN（P＜0.05），while the protein expression of cleaved-caspase-3 were decreased significantly （P＜0.05）. Compared with blank control ，the protein expression of YAP ，Bcl-2 and Beclin-1 were increased significantly after treated with 2.5％，5％LBJN（P＜0.05），while protein expression of cleaved-caspase- 3 and mTOR were decreased significantly （P＜0.05）. CONCLUSIONS ：LBJN can block the apoptosis of chondrocytes induced by YAP inhibitor verteporfin ，and its mechanism may be related to regulating the expression of apoptosis related proteins and enhancing autophagy of chondrocytes.
