In-Vitro α-amylase, α-glucosidase Inhibitory Activities and In-Vivo Anti-Hyperglycemic Potential of Different Dosage Forms of Guduchi (Tinospora Cordifolia [Willd.] Miers) Prepared With Ayurvedic Bhavana Process

Abstract

Guduchi (Tinospora cordifolia [Willd.] Miers) is a flagship rejuvenating herb of Ayurveda with reported anti-diabetic potential. In the present study, different dosage forms of Guduchi stem (growing on neem tree) were developed by adopting Ayurvedic pharmaceutical process of Bhavana (levigation). Guduchi Churna (GC) was subjected to 07 times Bhavana separately with its own extracted juice, decoction and potable water, and dosage forms namely Svarasa Bhavita Guduchi Churna (SBGC), Kwatha Bhavita Guduchi Churna (KBGC), and Jala Bhavita Guduchi Churna (JBGC) were prepared. The present study was aimed to evaluate the role of Bhavana on the potentiation of therapeutic properties of Guduchi. Sequential solvent extracts (5, 10, 15 and 25%) of GC, SBGC, KBGC and JBGC were prepared in different solvents [phosphate buffer, hexane, dichloromethane (DCM), chloroform] and screened for the α-amylase and α-glucosidase inhibitory activity. The results revealed that phosphate buffer and DCM extracts of SBGC exhibited strong α-amylase inhibitory potential (>80% inhibition at 25% concentration) followed by KBGC, JBGC and GC with reference to the standard acarbose. In α-glucosidase inhibitory activity, maximum inhibition was observed in DCM and chloroform extracts of SBGC (>85% inhibition at 25% concentration), followed by KBGC (>80% inhibition at 25% concentration), JBGC and GC. In vivo anti-hyperglycemic studies were carried out by oral glucose tolerance test in Swiss albino mice. Test drugs (JBGC, KBGC, SBGC) treated groups showed marginal decrease of blood glucose levels in normo glycemic mice. However, the blood glucose level in test drug JBGC, KBGC and SBGC treated groups was still within normal range in overnight fasted mice. In oral glucose tolerance test, among all dosage forms SBGC (51.08%) produced pronounced anti-hyperglycemic effect followed by KBGC (42.57%) at a dose of 520 mg/kg. The GC, JBGC, KBGC and SBGC samples were also standardized using berberine (a well established anti-diabetic compound) as a marker compound by HPTLC fingerprint analysis. Findings of the present study indicate that SBGC and KBGC can be used in the treatment of diabetes mellitus and gives supporting evidence to Ayurvedic claims that the Bhavana process has pharmaceutico-therapeutic significance in Ayurvedic drug development.

Keywords: anti-hyperglycemic; bhavana; guduchi; hypoglycemic; tinospora cordifolia; α-amylase; α-glucosidase.

FIGURE 1

Unit operating process of Svarasa Bhavita Guduchi Churna (SBGC), Kwatha Bhavita Guduchi Churna (KBGC), and Jala Bhavita Guduchi Churna (JBGC) samples preparation by traditional Bhavana method. (A)- (1) fresh Guduchi stems (2) chopped/crushed, (B)- (1) expressed juice (for SBGC batch) (2) decoction was prepared (for KBGC batch), (C)- wet grinding of Guduchi Churma with liquid media (Guduchi juice for SBGC batch, Guduchi decoction for KBGC batch, and potable water for JBGC batch) in edge-runner, (D)- soft and fine mass formed during grinding, (E)- observation of Subhavita Lakshana (Confirmatory tests for completion of levigation), (F)- wet granulation, (G)- hot air over drying, (H)- dry granules, (I)- preparation of tables.

FIGURE 2

α-amylase inhibitory activity of Guduchi Churna (GC), Svarasa Bhavita Guduchi Churna (SBGC), Kwatha Bhavita Guduchi Churna (KBGC), and Jala Bhavita Guduchi Churna (JBGC). (A) In phosphate buffer solution (PBS) (B) In dichloro methane (DCM) (C) In chloroform (D) In hexane.

FIGURE 3

α-glucosidase inhibitory activity of Guduchi Churna (GC), Svarasa Bhavita Guduchi Churna (SBGC), Kwatha Bhavita Guduchi Churna (KBGC), and Jala Bhavita Guduchi Churna (JBGC). (A) In phosphate buffer solution (PBS) (B) In dichloro methane (DCM) (C) In chloroform (D) In hexane.

FIGURE 4

Multivariate statistical analysis of codified different polarity Guduchi samples (Ph: phosphate buffer solution, D: dichloro methane, Ch: chloroform, and H: hexane) based on an unsupervized PCA model. (A) Unsupervized PCA score plot (B) Loading plot of α-amylase inhibitory activity of GC, JBGC, KBGC, SBGC formulations and various concentrations.

FIGURE 5

Multivariate statistical analysis of codified different polarity Guduchi samples (Ph: phosphate buffer solution, D: dichloro methane, Ch: chloroform, and H: hexane) based on an unsupervized PCA model. (A) Unsupervized PCA score plot and (B) Loading plot of α-glucosidase activity of GC, JBGC, KBGC, SBGC formulations and various concentrations. The ellipse represents the Hotelling T₂ with 95% confidence.

FIGURE 6

Effect of Svarasa Bhavita Guduchi Churna (SBGC), Kwatha Bhavita Guduchi Churna (KBGC), and Jala Bhavita Guduchi Churna (JBGC) on BSL in normal fasted mice.

FIGURE 7

Effect of Svarasa Bhavita Guduchi Churna (SBGC), Kwatha Bhavita Guduchi Churna (KBGC), and Jala Bhavita Guduchi Churna (JBGC) on BSL in glucose overloaded mice.

FIGURE 8

TLC plate scanned at 254 nm showing berberine chloride hydrate in the 3D samples chromatogram.