The Oral Administration of Sanguisorba officinalis Extract Improves Physical Performance through LDHA Modulation

Abstract

Muscle fatigue is induced by an acute or chronic physical performance inability after excessive physical activity often associated with lactate accumulation, the end-product of glycolysis. In this study, the water-extracted roots of Sanguisorba officinalis L., a herbal medicine traditionally used for inflammation and diarrhea, reduced the activities of lactate dehydrogenase A (LDHA) in in vitro enzyme assay myoblast C2C12 cells and murine muscle tissue. Physical performance measured by a treadmill test was improved in the S. officinalis-administrated group. The analysis of mouse serum and tissues showed significant changes in lactate levels. Among the proteins related to energy metabolism-related physical performance, phosphorylated-AMP-activated protein kinase alpha (AMPKα) and peroxisome proliferator-activated receptor-coactivator-1 alpha (PGC-1α) levels were enhanced, whereas the amount of LDHA was suppressed. Therefore, S. officinalis might be a candidate for improving physical performance via inhibiting LDHA and glycolysis.

Keywords: LDHA; Sanguisorba officinalis L.; glycolysis; herbal medicine; physical performance.

Figure 1

High-performance liquid chromatography (HPLC) analysis of S. officinalis (SO) extract. (A, B) The HPLC chromatogram of S. officinalis extract was monitored at ultraviolet detectors of 254 (A) and 203 nm (B). Representative compounds of S. officinalis extract were indicated by arrow lines.

Figure 2

Administration of S. officinalis (SO) extract improved physical performance. (A) Scheme of treatment and physical activity schedule. (B–D) The endurance performance (B), body weight (C), water intake (D), and food intake (E) of each group were measured. (F) Representative H&E staining of the quadriceps muscle tissue. The results of (B) are shown as mean ± SEM. *** p < 0.001 compared with group C of 3 weeks. # p < 0.05, ## p < 0.01, and ### p < 0.001 compared with group C of 4 week.

Figure 3

Measurement of physical performance-related factors. Serum levels of lactate (A), lactate dehydrogenase (LDH) (B), glucose (C), creatinine (D), and (E) aspartate transaminase (AST) and alanine aminotransferase (ALT) were measured by a commercially available assay kit. The results are shown as mean ± SEM. * p < 0.05 and *** p < 0.001 compared to the N group. # p < 0.05, ## p < 0.01, and ### p < 0.001 compared with group C. N; normal, C—control, P—positive control, L—low-dose S. officinalis, H—high-dose S. officinalis.

Figure 4

The administration of S. officinalis extract modulates physical performance-related factors in murine skeletal muscle. (A–C) The expressions of phospho-AMPK α (p-AMPKα), phosphorylated-AMP-activated protein kinase alpha (AMPKα), PGC-1α, and lactate dehydrogenase A (LDHA) were evaluated in the quadriceps muscle tissue. GAPDH and AMPKα expressions were used as control. Densitometry was measured with protein expression. The results are shown as mean ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with group N. # p < 0.05 and ### p < 0.001 compared with group C. N—normal, C—control, P—positive control, L—low-dose S. officinalis, H—high-dose S. officinalis.

Figure 5

The mode-of-action of S. officinalis (SO) (A) The protein expression of LDHA, Hsp90, CV-ATP5A, CIII-UQCRC2, CII-SDHB, CI-NDUFB8, and TOM20 were measured by Western blot analysis. Hsp90 expression was used as a control. (B) The activity of LDHA was measured by in vitro LDHA assay using recombinant human LDHA in the presence of an indicated concentration of S. officinalis extract. The results were shown as mean ± SEM. *** p < 0.001 compared to the control. ### p < 0.001 compared to the negative control. Analysis of extracellular acidification rate (ECAR) (C) and oxygen consumption rate (OCR) (D) was performed using a Seahorse XF analyzer to assess glycolysis and mitochondrial respiration, respectively. (E) Schematic representation of inhibition of S. officinalis extract on the LDHA expression and activity and subsequently enhancing the physical performance was illustrated.