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ORIGINAL ARTICLE
Year : 2021  |  Volume : 11  |  Issue : 1  |  Page : 80-87

HHOM-051608 A Whey Protein Formulation Fortified with Herbal Actives Improves Exercise Performance and Muscle Strength: A Preclinical Study


Research and Development, The Himalaya Drug Company, Bengaluru, Karnataka, India

Date of Submission09-Jun-2020
Date of Decision15-Jul-2020
Date of Acceptance28-Sep-2020
Date of Web Publication12-Feb-2021

Correspondence Address:
Mohamed Rafiq
Head-Discovery Sciences Group, Research and Development Center, The Himalaya Drug Company, Makali, Bengaluru-562 162, Karnataka
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ijnpnd.ijnpnd_50_20

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   Abstract 


Context: The lifestyle of individuals is changing rapidly in today’s world and physical fitness is an essential requirement to keep oneself fit and healthy. Nutritional supplements could improve physical fitness by modulating energy metabolism, oxidative processes, and perceived fatigue. Aim: This study aimed to evaluate the effects of HHOM-051608, which is a first-of-its-kind whey protein supplement, packed with naturally rich essential amino acids and herbal ingredients to improve the exercise performance, energy metabolism, and physical endurance. Setting and study design: Thirty-two male Wistar rats were divided into four groups of eight each. Group 1 served as sedentary control; group 2 was sedentary rats treated with HHOM-051608 (2 g/kg b.w.); group 3 was exercise training control, and group 4 was exercise training rat treated with HHOM-051608 (2g/kg b.w.). Methods: The exercise training rats were trained on a motorized rat treadmill at a constant speed for 8 weeks. After 8 weeks, all animals were subjected to a grip strength test followed by a treadmill endurance test. Statistical analysis: The results were statistically analyzed by one-way analysis of variance followed by Tukey comparison test, using Prism GraphPad 6.07 (GraphPad Software Inc, San Diego, CA, USA). A P value < 0.05 was considered statistically significant. Results: HHOM-051608 supplemented animals showed a significant increase in grip strength and run time on the treadmill. Conclusion: HHOM-051608 has improved the exercise performance, muscle strength, and muscle glycogen. Thus can be recommended to enhance exercise performance, build muscle mass and decrease muscle soreness.

Keywords: Exercise performance, grip strength, herbal actives, physical endurance, whey protein


How to cite this article:
Vishwakarma KK, Narendra MM, Azeemuddin MM, Shankar P, Babshet P, Rafiq M, Babu UV. HHOM-051608 A Whey Protein Formulation Fortified with Herbal Actives Improves Exercise Performance and Muscle Strength: A Preclinical Study. Int J Nutr Pharmacol Neurol Dis 2021;11:80-7

How to cite this URL:
Vishwakarma KK, Narendra MM, Azeemuddin MM, Shankar P, Babshet P, Rafiq M, Babu UV. HHOM-051608 A Whey Protein Formulation Fortified with Herbal Actives Improves Exercise Performance and Muscle Strength: A Preclinical Study. Int J Nutr Pharmacol Neurol Dis [serial online] 2021 [cited 2021 Mar 9];11:80-7. Available from: https://www.ijnpnd.com/text.asp?2021/11/1/80/309282




   Introduction Top


The popularity of functional foods and nutraceuticals has increased in the modern world, which has driven focus on research that supports and suggests the health benefits of nutraceutical products.[1] The nutritional dietary supplement products usually consumed with routine diet are commercially available worldwide. Earlier, these nutritional supplements were used mostly by athletes to stimulate and maintain muscle growth but the horizon of nutritional supplements has increased to almost all age groups and various diseases and disorders.[2],[3],[4],[5] Many individuals involved in bodybuilding and athletes use protein supplements for muscle growth and strength and to increase the release of growth hormone. As per literature, the athlete should consume approximately 12% to 15% of their daily total energy intake as protein, equivalent to about 1.5 to 2.0 g protein/kg per day.[6]

The main sources of protein easily available are casein, whey, or soy proteins, and among these, whey is the most preferred one. Whey protein comprises about 20% of the total bovine milk protein and consists mainly of 7-lactoglobulin (50%), f3-lactalbumin (25%), serum albumin (7%), and immunoglobulins (5%). Whey protein contains high concentrations of branched-chain amino acid (BCAA) compounds that are involved in protein synthesis.[7] BCAAs are sources of nitrogen for the synthesis of non-essential amino acids such as glutamine and alanine.[8] Leucine is a potent inhibitor of muscle protein degradation that enhances insulin secretion[9] and may also be an important precursor for gluconeogenesis in both liver and muscle.[10] It has been shown that dietary BCAA augments exercise-induced mitochondrial biogenesis[11] and energy metabolism. Protein ingested with an optimal amount of carbohydrates potentiates post-exercise glycogen recovery particularly when co-administered.[12] BCAA-enriched essential amino acids are shown to attenuate post-exercise muscle soreness and enhance muscle repair in rats,[13] and it is effective in alleviating only low-to-moderate extent of muscle damage.[14] One-third of ingested carbohydrates are taken up by skeletal muscle, where it is either stored as glycogen or metabolized to lactate or CO2 and H2O. Depletion of muscle glycogen stores has been associated with fatigue during constant-load exercise,[15] and it is known that the storage levels are increased or speedily recovered with dietary supplements.

HHOM-051608, a proprietary herbal, whey protein blend of The Himalaya Drug Company, Bangalore, was formulated considering the beneficial aspects of the ingredients mentioned above. It is a blend of three forms of whey, namely whey protein concentrate, whey protein isolate, and whey protein hydrolysate. It is enriched with herbal ingredients like Withania somnifera (WS), Cissus quadrangularis (CQ), and Punica granatum (PG), which are commonly known in India as Ashvagandha, Hadjod, and Pomegranate, respectively, and also with high-quality proteins and nutrients such as vitamins, minerals, taurine, and calcium. The major ingredients of the supplement are whey protein blend (82.5%, whey protein concentrate, whey protein isolate, and whey protein hydrolysate), PG juice powder (0.61%), CQ (0.60%), and WS (0.10%). The objective of the present study was to assess the effect of HHOM-051608 on exercise performance, energy metabolism, and physical endurance of rats.


   Materials and Methods Top


This study is of preventive design; the animals were supplemented with HHOM-051608 along with the exercise for 4 weeks before subjecting for a physical endurance test. Inbred male Wistar rats were housed in standard conditions of temperature (22 ± 3°C), relative humidity (55 ± 5%), and light (12 hour light/dark cycle) before and during the study. They were fed ad libitum with standard rat pellet feed manufactured by VRK Nutritional Solutions, Pune, India. Deep bore-well water passed through Aquaguard Reviva reverse osmosis plant manufactured by Eureka Forbes Ltd., Mumbai, India was provided ad libitum. All the experimental protocols were approved by the Institutional Animal Ethics Committee (IAEC) of The Himalaya Drug Company, Bangalore, and the animals received humane care as per the guidelines prescribed by the Committee for the Purpose of Control and Supervision on Experiments on Animals (CPCSEA), The Ministry of Environment & Forests, Government of India.

Procedure

Thirty-two male Wistar rats weighing 250 to 280 g, were divided into four groups of eight each. Group 1 served as sedentary control (Ctrl/Sed), group 2 as sedentary group treated with HHOM-051608 at a dose of 2 g/kg b.w. (HHOM-051608/Sed), group 3 as exercise training control (Ctrl/Ex), and group 4 as exercise training group treated with HHOM-051608 at a dose of 2 g/kg b.w. (HHOM-051608/Ex).

Stimulus exercise

Rats in the exercise-trained groups were trained to run on a motor-driven treadmill for 10 minutes with a daily gradual increase in treadmill speed up to 15 m/min during the acclimatization period of 5 days. After acclimatization, animals were subjected to actual training for 8 weeks with a running speed of 15 m/min at 5% inclination for 30 min/day and 5 days/week.[16],[17] The treatment was started from fifth week of the study and continued till eighth week. Weekly body weight and food consumption were recorded.

Grip strength test

At the end of the eighth week, a grip strength test was performed. Rats were placed with their forepaws on a grill, and they were gently pulled backward until they released their grip.[18] A grip meter (Bio-GT3, BIOSEB, France), attached to a force transducer, measured the peak force generated. A total of three trials were performed with each animal and the results were expressed as the mean of three values in Newtons (N).

Endurance test

After the grip strength test, all rats were subjected for an endurance test on a treadmill with a speed of 25 m/min until exhaustion. During the treadmill test, the rats started spending more and more time in the shock zone as they get exhausted. Rats spending more than 30 seconds in this zone were considered as fully exhausted, and were removed from the treadmill. The run time, speed, and other parameters were recorded for each rat using these criteria.

Blood and tissue collection

Blood was collected from each rat immediately after exhaustion using isoflurane anesthesia. The blood samples were centrifuged at 3000g for 15 minutes and the serum samples were stored at −80°C. After blood collection, rats were euthanized using deep isoflurane anesthesia. Gastrocnemius muscle, soleus muscle, and epididymal fat tissues were quickly excised, washed with saline, weighed, and stored at −80°C for further analysis.

Serum glucose, serum creatine kinase, total protein and lipid profile

Serum glucose, serum creatine kinase, total protein and lipid profile were measured by an auto-analyzer using standard kits from ERBA (Erba EM-360 from Transasia Biomedicals, Mumbai).

Muscle glycogen

The muscle glycogen was measured as described by Folin’s method.[19] Briefly, 100 mg of freshly collected gastrocnemius muscle was added to 2 mL of 30%, w/v, potassium hydroxide, and 2.5 mL of absolute alcohol was added to precipitate the glycogen and centrifuged at 2000 rpm for 15 minutes. The supernatant was discarded and the dried pellet was reconstituted with 1 mL of distilled water to make a glycogen stock solution. Phenol (80%) and concentrated sulphuric acid were added under ice-cold condition. The absorbance of the resulting solution was measured at 490 nm. The muscle glycogen content was derived using standard glycogen concentration curve.

Statistical analysis

All values were expressed as the mean ± standard error of mean. The results were statistically analyzed by one-way analysis of variance followed by Tukey comparison test, using Prism GraphPad 6.07 (GraphPad Software Inc, San Diego, CA, USA). A P value < 0.05 was considered statistically significant.


   Results Top


Effect of HHOM-051608 on body weight and feed consumption

Changes in body weight and average feed consumption are presented in [Figure 1] and [Figure 2]. The sedentary group of rats showed a marginal increase in body weight compared to trained rats. The feed consumption of the trained rats decreased compared to the sedentary rats and not much change was observed in HHOM-051608 treated group compared to Ctrl/Ex group.
Figure 1 Effect of HHOM-051608 on body weight of sedentary and exercised rats after 4 weeks of supplementation

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Figure 2 Effect of HHOM-051608 on feed consumption of sedentary and exercised rats after 4 weeks of supplementation

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Effect of HHOM-051608 on endurance and grip strength

A significant increase in physical endurance (P ≤ 0.001) and strength (P ≤ 0.05) was observed in the HHOM-051608/Ex group compared to Ctrl/Ex group [Figure 3] and [Figure 4].
Figure 3 Effect of HHOM-051608 on the endurance of sedentary and exercised rats after 4 weeks of supplementation

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Figure 4 Effect of HHOM-051608 on grip strength of sedentary and exercised rats after 4 weeks of supplementation

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Effect of HHOM-051608 on muscles and epididymal fat weight

Sedentary rats and exercise-trained rats treated with HHOM-051608 showed an appreciable increase in relative gastrocnemius muscle weight compared to their respective control groups. Relative soleus muscle weight was also found to be higher in HHOM-051608/Ex group when compared with Ctrl/Ex group. A significant decrease in epididymal fat was found in HHOM/Ex (P ≤ 0.05) group compared to Ctrl/Ex group [Table 1].
Table 1 Effect of HHOM-051608 on muscle weight and epididymal fat of sedentary and exercised rats

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Effect of HHOM-051608 on muscle glycogen

No change was observed in the muscle glycogen in the case of HHOM-051608/Sed rats when compared to Ctrl/Sed rats. However, a marginal increase was observed in the HHOM-051608/Ex rats when compared to Ctrl/Ex group [Figure 5].
Figure 5 Effect of HHOM-051608 on muscle glycogen of sedentary and exercised rats after 4 weeks of supplementation

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Effect of HHOM-051608 on serum biochemical parameters

The serum glucose level was higher in HHOM-051608/Ex group and a significant (P ≤ 0.05) decrease in serum cholesterol was observed in HHOM-051608 treated groups compared to Ctrl/Ex group. No significant change was observed in triglyceride and high density lipoprotein (HDL) levels [Table 2]. Plasma concentrations of creatine kinase (CK) and lactate dehydrogenase (LDH) increased with exercise; supplementation with “HHOM-051608” decreased serum CK and LDH concentrations compared with exercise control. However, no much change was observed in the serum total protein level in HHOM-051608 treated groups compared to the respective control groups.
Table 2 Effect of HHOM-051608 on biochemical parameters of sedentary and exercised rats

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   Discussion Top


Animal proteins are an important source of protein, however, potential health concerns do exist when consumed primarily from animal sources. With a proper combination of sources, vegetable proteins may provide similar benefits as proteins from animal sources. Whey is one of the rich sources of complete protein whose biologically active components provide additional benefits to enhance human function. It contains amino acid cysteine, which appears to enhance glutathione levels, which has been shown to have strong antioxidant properties that can assist the body in combating various diseases. Whey protein also contains a high concentration of BCAA that is important for tissue maintenance and prevention of catabolic actions during exercise.[20] Whey protein supplementation suppresses hunger when compared with casein or soy and egg albumin. In agreement with the previous findings, our study showed similar results where the supplementation of HHOM-051608 decreased the body weight and feed intake of rats. This may be due to the presence of BCAA, especially leucine, which acts on central receptors and signal anorexigen peptide synthesis, probably via mTOR.[21]

Forelimb grip strength is one of the routine physical examinations to assess muscle strength.[22] A significant increase (P<0.05) was found in the grip strength of the animals treated with HHOM-051608. This may be due to whey protein, which could improve muscle strength because of its amino acid composition and also due to the presence of WS, which is known to increase muscle strength and recovery.[23],[24]

The effect of exercise training (ET) and/or HHOM-051608 supplementation on improving exercise endurance capacity was evaluated with a treadmill running test. All animals underwent a run-to-exhaustion exercise test at moderate treadmill speed. It was found that HHOM-051608 supplementation could prolong the exhaustion time significantly after 8 weeks of exercise training. Previous reports have demonstrated that long-term ET can cause an imbalance in antioxidant status and result in acute tissue injury or muscle fatigue that causes muscle damage and affects protein metabolism in the muscle.[25] HHOM-051608 supplementation after ET reduced the resulting physiologic fatigue, thereby contributing a significant (P < 0.001) improvement in exercise performance. This effect may be due to PG extract that is known for its anti-oxidant and rejuvenating properties and it is also reported to accelerate recovery of muscle damage and soreness that are very important factors to be considered in resistant exercise.[26],[27],[28]

Furthermore, many studies reported that BCAA is known to promote protein synthesis.[29] The daily supplementation with whey protein in combination with ET was found effective in promoting muscle hypertrophy.[30] In the present work, 8 week aerobic running ET has increased gastrocnemius and soleus muscle weight, which has further increased with HHOM-051608 supplementation. This may be due to the BCAA present in HHOM-051608, especially leucine, which plays an important role in protein synthesis.[31] Moderate to high-intensity aerobic exercise could reduce visceral adipose tissue in overweight subjects.[32] Exercise could be a new physiological stimulus in counteracting obesity by providing adrenergic-regulated transdifferentiation of white-to-brown adipocytes.[33] In the present study, it was observed that HHOM-051608 treated animals showed a significant (P < 0.05) decrease in epididymal fat pad compared to control rats.

ET is known to be an effective method in reducing fat mass and increasing lean body mass via improving lipid–lipoprotein profile[34] and insulin sensitivity.[35] In the present work, we found that ET rats treated with HHOM-051608 significantly decreased the total cholesterol level compared to exercise control rats.

An intensive exercise or long-term training could elevate biochemical markers, LDH, CK, and total protein, of post-exercise cardiac or skeletal muscle damage,[22] which remains elevated up to 24 hours post-workout.[36] High protein supplementation is known to attenuate muscle strength and LDH levels in animal models.[37] In this study, a significant elevation in serum LDH and CK levels that are the markers of tissue damage and strenuous exercise decreased with the supplementation of HHOM-051608, which showed its protective effect against muscle injury and soreness.

Based on the above results, it can be inferred that supplementation of HHOM-051608 increased the stamina, strength, and anabolic muscle mass of exercised rats after 4 weeks of supplementation. It was also found to be effective in decreasing blood cholesterol and correcting the lipid profile. This showed supplementation of HHOM-051608 along with exercise achieved better results compared to exercise control.

The ability of whey supplementation to improve muscle strength is of great benefit to sportspersons who would like to maintain an appropriate lipid profile, low adiposity, increased muscle strength, and enhanced performance. It is useful as additional quick-absorbing, good quality protein for adults whose protein intake is inadequate through their regular meals. Whey proteins also help those who are recovering from illnesses, in convalescence, and require additional proteins to have a better immune response and faster recovery. Additionally, whey proteins also help in improving muscle strength in the elderly and those who have lower muscle strength due to prolonged immobility or illness. The additional value of herbs (WS, CQ, and PG) helps in aiding muscle mass, increasing bone health, and shortens recovery time.

HHOM-051608, which is a combination of both whey protein and herbs, can be recommended to enhance the exercise performance, build muscle mass, and decrease muscle soreness; consuming this during the training period will have immense potential to raise performance to the next level. Thus, HHOM-051608 would be an ideal protein supplement for fitness enthusiasts. This product is high in proteins and hence, caution and medical advice are necessary before recommending for a patient with compromised kidney function.

Acknowledgment

The authors acknowledge Functional foods, Natural Product Innovations, and Preclinical Pharmacology and Toxicology Departments, R&D Center, The Himalaya Drug Company, for providing support during the study. The authors also acknowledge M/S The Himalaya Drug Company, for providing the facilities, funds, and support for performing this research work.

Financial support and sponsorship

This work was funded by The Himalaya Drug Company Bangalore.

Conflicts of interest

The authors listed are the employees of The Himalaya Drug Company, Bangalore, India when the research work was carried out. The authors declare no other conflicts of interest.



 
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    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5]
 
 
    Tables

  [Table 1], [Table 2]



 

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