Evaluation of Renal and Cardioprotective Potential of the Biofield Energy Treated Proprietary Test Formulation on L-NAME and High Fat Diet-Induced Cardiovascular Disorders in Sprague Dawley Rats

The aim of this study was to evaluate the impact of Biofield Energy Treated/Blessed Proprietary Test Formulation and Biofield Energy Treatment/Blessing per se on kidney biomarkers on L-NAME and high fat diet (HFD)-induced cardiovascular disorders in Sprague Dawley rats. In this experiment, the functional kidney biomarkers such as epinephrine/adrenaline, inducible nitric oxide synthase (iNOS), angiotensin-II, C-reactive protein (CRP), and renin were measured using ELISA assay. A test formulation was formulated including minerals (magnesium, zinc, copper, calcium, selenium, and iron), vitamins (vitamin C, vitamin B6, vitamin B12, vitamin B9, and vitamin D3), cannabidiol (CBD) isolate, Panax ginseng extract, and β-carotene. The components of the test item were divided into two; one section was defined as the untreated test formulation, while the other part and three group of animals received Mr. Mahendra Kumar Trivedi’s Biofield Energy healing/Blessing remotely for about 3 minutes. The results showed that the level of adrenaline was reduced by 31.62%, 19.58%, 34.32%, 37.07%, and 29.87% in the G5 (L-NAME + HFD + the Biofield Energy Treated test formulation), G6 (L-NAME + HFD + Biofield Energy Treatment per se to animals from day -15), G7 (L-NAME + HFD + the Biofield Energy Treated test formulation from day-15), and G8 (L-NAME + HFD + Biofield Energy Treatment per se plus the Biofield Energy Treated test formulation from day-15), and G9 (L-NAME + HFD + Biofield Energy Treatment per se animals plus the untreated test formulation) groups, respectively as compared to the disease control group (G2). Moreover, the level of iNOS was reduced by 56.76%, 49.51%, 61.79%, 57.63%, and 62.44% in the G5, G6, G7, G8, and G9 groups, respectively, as compared to the disease control group (G2). Additionally, the level of angiotensin-II was decreased by 41.09%, 34.92%, 60.65%, 53.28%, and 60.09% in the G5, G6, G7, G8, and G9 groups, respectively, as compared to the G2 group. The level of CRP was decreased by 47.21%, 38.89%, 59.81%, 55.52%, and 64.02% in the G5, G6, G7, G8, and G9 groups, respectively as compared to the G2 group. Besides, the level of renin was decreased by 20.27%, 20.13%, 12.99%, and 25.73% in the G5, G7, G8, and G9 groups, respectively as compared to the G2 group. Overall, the data suggested significance improvement of vital functional kidney biomarkers of the Biofield Energy Treated/ Blessed test formulation and Biofield Energy Treatment per se along with preventive measure on the animal with respect to various pathological conditions that might be beneficial various types of cardiovascular disorders. Therefore, the results showed the significant slowdown the inflammation-related cardiovascular disease progression and its complications/symptoms in the preventive Biofield Energy Treatment group per se and/or Biofield Energy Treated/Blessed Test formulation groups (viz. G6, G7, G8, and G9). DOI: 10.14302/issn.2574-4488.jna-21-3847 Freely Available Online www.openaccesspub.org | JNA CC-license DOI : 10.14302/issn.2574-4488.jna-21-3847 Vol-1 Issue 3 Pg. no.13 Corresponding author: Snehasis Jana, Trivedi Science Research Laboratory Pvt. Ltd., Thane (W), Maharashtra, India.


Introduction
Cardiovascular diseases (CVDs) are the leading cause of death worldwide. A randomized controlled trial indicates that epinephrine increases return of pulses for cardiac arrest patients. Despite increases in return of pulses, it reduces long-term survival and functional recovery after cardiopulmonary resuscitation (CPR). Laboratory data also suggest that it induced reductions in microvascular blood flow during and after CPR [1]. Nitric oxide (NO) is produced in almost all tissues and organs by 3 distinct NO synthase (NOS) isoforms (neuronal, inducible, and endothelial NOS), all the enzymes are expressed in the human cardiovascular system [2]. Abnormal generation of NO is considered as a major cause of coronary heart disease (CHD). It has been shown that endothelial dysfunction is characterized by reduced endothelial NO synthesis by constitutive NOS (cNOS) and increased systemic NO synthesis due to increased iNOS activity can leads to cardiovascular disorders [3]. Angiotensin II is considered one of the important mediator of the renin-angiotensin system (RAS). It has been reported that angiotensin-II plays a vital role for the pathophysiology of cardiovascular disorders such as hypertension, atherosclerosis, coronary heart disease, restenosis, and heart failure through the RAS [4,5].
C-reactive protein (CRP) seems to predict the risk of cardiovascular problems as well as cholesterol levels. A recent study reported that elevated levels of CRP is associated with three-times more risk of heart attack.
CRP is one of the best possible marker of vascular inflammation and plays a vital role in promoting vascular inflammation, vessel damage and clinical cardiovascular disease [6,7]. Evidence from preclinical and clinical studies suggests that renin inhibitors may have renal and cardiovascular effects [8]. Thus provide vital physiological roles [9][10][11]. Besides, cannabidiol itself has wide range of pharmacological profile and has been reported to role in different disorders [12,13], while ginseng extract is regarded as the one of the best immune booster for overall immunity [14]. The animals in groups G2-G9 were received L-NAME (20 mg/kg, i.p.) and a HFD throughout the experimental period. At the end of the experimental period (8 weeks treatment), the animals were sacrifice, remove kidney, homogenate and subjected for the estimation of epinephrine, iNOS, angiotensin-II, CRP, and renin.

Estimation of Different Biomarkers in Kidney Homogenate
The kidney homogenate from all the groups was subjected for the estimation of level of various vital biomarkers such as epinephrine, iNOS, angiotensin-II, CRP, and renin. All the biomarkers were estimated using ELISA assay as per manufacturer's procedure. The assay is a quantitative method and the principle based on the binding of antigen and antibody in sandwich manner.

Estimation of Epinephrine/Adrenaline in Kidney Homogenate
The expression of adrenaline in the kidney homogenate was measured in the presence of the test formulation and the data are shown in Figure 1.

Estimation of iNOS Kidney Tissue Homogenate
The effect of the test formulation and Biofield Energy Treatment per se on the expression of induced nitric oxide synthase (iNOS) is shown in Figure 2. The disease control (L-NAME + high fat diet (HFD) + 0.5% CMC) group (G2) showed value of iNOS as 88. 36   mechanism of endothelial dysfunction, and that causes cardiovascular abnormalities [30,31]. Besides, iNOS is expressed due to the effects of proinflammatory cytokines and can release more NO than other isoform of nitric oxide synthase enzymes [32]. Overall, in this study the Biofield Energy Treated test formulation and Biofield Energy Treatment per se significantly reduced the level of iNOS, which was increased due to cardiovascular disease condition, induced by L-NAME and HFD that could be beneficial in the cardiovascular patients

Estimation of Angiotensin-II in Kidney Homogenate
The level of angiotensin-II in kidney homogenate was measured and the data are shown in      Figure 4). Inflammation plays a major role in the pathogenesis of cardiovascular disease [35]. In this context, CRP is plays an independent risk factor for cardiovascular patients and one of the best biomarker for detection of immune function alterations [36,37]. Therefore, in this experiment the Biofield Energy Treated test formulation and Biofield Energy Treatment per se reduced the level of CRP, which could be beneficial to improve the cardiovascular disease conditions.

Estimation of Renin in Kidney Tissue
The effect of the test formulation and Biofield Energy Treatment per se on the level of renin in kidney tissue and the results are shown in Figure 5. The level of renin in the disease control (L-NAME + high fat diet,