Affiliations: [a] Department of Pharmacology, Institute of Basic Medical Sciences, Khyber Medical University, Peshawar, KPK, Pakistan
| [b] Department of Pharmacy, Women Institute of Learning (WIL), Abbottabad, KPK, Pakistan
| [c] Department of Pharmacy, Quaid-i-Azam University, Islamabad, Pakistan
| [d] Department of Pharmacy, Sarhad University of Science and Information Technology, Peshawar, KPK, Pakistan
| [e] Khyber Medical College, Khyber Medical University, Peshawar, KPK, Pakistan
| [f] Faculty of Pharmacy, Gomal University, Dera Ismail Khan, KPK, Pakistan
Correspondence:
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Corresponding author: Kifayat Ullah Shah, Department of Pharmaceutics, Faculty of Pharmacy, Gomal University, D.I. Khan, 23431, Khyber Pakhtunkhwa, Pakistan. Tel.: +92 3349008556; E-mail: kifayatrph@gmail.com.
Abstract: Diarrhea is a major health problem and despite of interventions, it remains a substantial cause of mortality and morbidity. In this study, a flavonoids-rich Vitex negundo extract was mechanistically evaluated for its effectiveness in diarrheal diseases. The Vitex negundo ethanolic extract was subjected to extraction for isolation of total flavonoids and qualitative phytochemical analysis. The acute toxicity of flavonoids-extract was done in mice to assess its safety and tolerability. The anti-diarrhoeal activity was determined using the castor oil induced diarrhea mouse model at doses of 10, 30 and 100 mg/kg in relation to loperamide (10 mg/kg, oral). The antispasmodic profile was determined by using cut pieces of rabbit’s jejunum in Tyrode’s solutions, employing acetylcholine (0.03μM) as a tissue stabilizer and verapamil, as a calcium channel blocker. The preliminary qualitative analysis of extract revealed different phytochemicals, in addition to flavonoids. The acute toxicity profile showed that the flavonoids-extract is safe and tolerable (LD50 = 1678 mg/kg). In the tissue experiments, the total flavonoids exhibited an EC50 of 1.52 mg/mL and showed maximum relaxation of spontaneous contractions at 5.0 mg/mL and against high-K+ induced contractions at 3 mg/mL (EC50 = 0.43 mg/mL). The spasmolytic activity of total flavonoids was comparable to verapamil which suggests that the activity might be due to the blockade of calcium channels. The flavonoids extract (0.1, 0.3 and 1.0 mg/mL) produced an adequate right shift in the calcium concentration response curve as compared to the control (EC50 value = –2.67 mg/mL), which confirmed that the extract has calcium channel blocking activity.
Keywords: Flavonoids and diarrhea, spasmolytic, flavonoids-rich Vitex negundo
, calcium channels antagonist, mechanism based anti-diarrheal therapy
