Effects of phased joint intervention on Rho/ROCK expression levels in patients with portal hypertension

Abstract

The current study investigated the effects of phased joint intervention on clinical efficacy and Rho/Rho-associated coil protein kinase (ROCK) expression in patients with portal hypertension complicated by esophageal variceal bleeding (EVB) and hypersplenism. Patients with portal hypertension (n=53) caused by liver cirrhosis complicated by EVB and hypersplenism treated with phased joint intervention were assessed, and portal hemodynamics, blood, liver function, complications, and rebleeding incidence were analyzed. Reverse transcription-quantitative polymerase chain reaction was used to measure Rho, ROCK1 and ROCK2 mRNA expression levels in peripheral blood mononuclear cells prior to and following phased joint intervention, and western blotting was employed to determine the protein expression levels of Rho, ROCK1, ROCK2, phosphorylated (p) myosin phosphatase target subunit 1 (MYPT1) and total-MYPT1. All patients underwent an emergency assessment of hemostasis with a 100% success rate. Varicose veins were alleviated, and portal hemodynamics and liver function improved following intervention. Furthermore, preoperative and postoperative expression levels of Rho, ROCK1 and ROCK2 mRNA were higher compared with the control group. Notably, the mRNA expression levels of Rho, ROCK1 and ROCK2 in the postoperative group were significantly lower when compared with the preoperative group. Protein expression levels of Rho, ROCK1, ROCK2 and pMYPT1 in the postoperative group were lower, as compared with the preoperative group. Concentration levels of transforming growth factor-β1, connective tissue growth factor and platelet-derived growth factor in peripheral blood were significantly reduced following phased joint intervention. Therefore, the present findings demonstrated that phased joint intervention is able to effectively treat EVB and hypersplenism, and improve liver function. The efficacy of phased joint intervention may be associated with its role in the regulation of the Rho-ROCK signaling pathway.

Keywords: Rho; cirrhosis; embolization; esophageal varices; rock.

Figure 1.

Changes in esophageal varices prior to and after treatment, demonstrated by digital subtraction angiography (DSA). (A) Varicose vein as revealed by portal vein angiography prior to intervention. (B) Percutaneous transhepatic variceal embolization varicose vein occlusion following intervention. (C) Enlarged spleen demonstrated by DSA prior to partial splenic embolization (PSE) intervention. (D) Partial spleen embolization after PSE intervention. (E) Enlarged spleen embolization after repeat PSE intervention 3 months later.

Figure 2.

Rho, ROCK1 and ROCK2 expression levels before and after treatment, as determined by reverse transcription-quantitative polymerase chain reaction. mRNA expression levels of Rho, ROCK1 and ROCK2 were higher compared with the normal control group Prior to and after intervention (Rho: preoperative, 10.59±0.19 vs. 1.80±0.46; postoperative, 3.29±0.47 vs. 1.80±0.46; ROCK1: preoperative, 2.79±0.27 vs. 0.44±0.08; postoperative, 1.21±0.09 vs. 0.44±0.08; and ROCK2: preoperative, 3.48±0.19 vs. 0.65±0.20; postoperative, 1.84±0.11 vs. 0.65±0.20). mRNA expression levels of Rho, ROCK1 and ROCK2 in the postoperative group were significantly lower compared with those of the preoperative group (Rho: 3.29±0.47 vs. 10.59±0.19; ROCK1: 1.21±0.09 vs. 2.79±0.27; and ROCK2: 1.84±0.11 vs. 3.48±0.19). *P<0.001, **P=0.002 and ***P=0.004 vs. the control group; ^#P<0.001 vs. the preoperative group. ROCK, Rho/Rho-associated coil protein kinase.

Figure 3.

Protein expression levels of Rho, ROCK1 and ROCK2 prior to and after treatment, as determined by (A) western blotting and (B) densitometry. Protein expression levels were normalized to β-actin. 1, normal control group; 2, preoperative group; 3, postoperative group. Data are presented as the mean ± standard deviation. *P<0.05, **P<0.01 vs. the control group; ^#P<0.05, ^##P<0.01 vs. the preoperative group. ROCK, Rho/Rho-associated coil protein kinase; p phosphorylated; t, total; MYPT1, myosin phosphatase target subunit 1.

Figure 4.

TGF-β1, CTGF and PDGF concentrations in the peripheral blood of the preoperative and postoperative groups, determined by ELISA. TGF-β1, CTGF and PDGF concentrations in the peripheral blood following intervention were significantly reduced compared with the preoperative parameters (TGF-β1, 552.89±69.49 vs. 837.19±99.14; CTGF, 147.68±7.62 vs. 190.94±9.66; and PDGF, 454.30±57.74 vs. 638.74±66.68). *P<0.001 vs. the preoperative group. A, preoperative group; B, postoperative group; TGF, transforming growth factor; CTGF, connective tissue growth factor; PDGF, platelet-derived growth factor.